U.S. patent application number 17/280358 was filed with the patent office on 2021-11-25 for use of soluble trem-1 levels for identifying subjects susceptible to respond to an anti-inflammatory therapy.
This patent application is currently assigned to INOTREM. The applicant listed for this patent is INOTREM. Invention is credited to Amir BOUFENZER, Kevin CARRASCO, Marc DERIVE, Jean-Jacques GARAUD, Lucie JOLLY, Martin KOCH, Aurelie OLIVIER, Margarita SALCEDO-MAGGUILLI.
Application Number | 20210364532 17/280358 |
Document ID | / |
Family ID | 1000005799199 |
Filed Date | 2021-11-25 |
United States Patent
Application |
20210364532 |
Kind Code |
A1 |
DERIVE; Marc ; et
al. |
November 25, 2021 |
USE OF SOLUBLE TREM-1 LEVELS FOR IDENTIFYING SUBJECTS SUSCEPTIBLE
TO RESPOND TO AN ANTI-INFLAMMATORY THERAPY
Abstract
A method for identifying human subjects suffering from an
inflammatory disorder susceptible to respond to a therapy, in
particular to a TREM-1 inhibitor. In particular, an in vitro method
for identifying a human subject suffering from an inflammatory
disorder, preferably an acute inflammatory disorder such as SIRS,
sepsis or septic shock, susceptible to respond to a therapy, in
particular to a TREM-1 inhibitor. The method includes the steps of
a) measuring the level of soluble triggering receptors expressed on
myeloid cells-1 (sTREM-1) in a biological sample from the human
subject, comparing the level of sTREM-1 measured at step a) to a
predetermined sTREM-1 value, and c) identifying a human subject
suffering from an inflammatory disorder with a level of sTREM-1
measured at step a) higher than the predetermined sTREM-1 value of
step b) as susceptible to respond to a therapy, in particular to a
TREM-1 inhibitor.
Inventors: |
DERIVE; Marc;
(Villers-les-Nancy, FR) ; OLIVIER; Aurelie;
(Clairefontaine-en-Yvelines, FR) ; CARRASCO; Kevin;
(Dombasle-sur-Meurthe, FR) ; KOCH; Martin;
(Montauban, FR) ; SALCEDO-MAGGUILLI; Margarita;
(Chatillon, FR) ; BOUFENZER; Amir;
(Villers-les-Nancy, FR) ; JOLLY; Lucie;
(Bar-le-Duc, FR) ; GARAUD; Jean-Jacques; (Paris,
FR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
INOTREM |
Vandoeuvre-les-Nancy |
|
FR |
|
|
Assignee: |
INOTREM
Vandoeuvre-les-Nancy
FR
|
Family ID: |
1000005799199 |
Appl. No.: |
17/280358 |
Filed: |
September 27, 2019 |
PCT Filed: |
September 27, 2019 |
PCT NO: |
PCT/EP2019/076263 |
371 Date: |
March 26, 2021 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
G01N 2333/705 20130101;
G01N 33/6893 20130101; A61K 38/10 20130101; G01N 2800/52
20130101 |
International
Class: |
G01N 33/68 20060101
G01N033/68; A61K 38/10 20060101 A61K038/10 |
Foreign Application Data
Date |
Code |
Application Number |
Sep 28, 2018 |
EP |
18306277.7 |
Claims
1-15. (canceled)
16. A method for treating an inflammatory disorder in a human
subject identified as susceptible to respond to a therapy, said
method comprising: identifying a human subject suffering from an
inflammatory disorder susceptible to respond to a therapy by: a)
measuring the level of soluble Triggering Receptors Expressed on
Myeloid cells-1 (sTREM-1) in a biological sample from the human
subject; b) comparing the level of sTREM-1 measured at step a) to a
predetermined sTREM-1 value; c) identifying a human subject
suffering from an inflammatory disorder with a level of sTREM-1
measured at step a) higher than the predetermined sTREM-1 value of
step b) as susceptible to respond to a therapy; administering a
therapy to the subject suffering from an inflammatory disorder
identified as being susceptible to respond to a therapy.
17. The method according to claim 16, wherein the predetermined
sTREM-1 value of step b) is obtained from a reference
population.
18. The method according to claim 17, wherein the reference
population is a population of human patients suffering from an
inflammatory disorder.
19. The method according to claim 17, wherein the reference
population is a population of human patients suffering from
systemic inflammatory response syndrome (SIRS), sepsis or septic
shock.
20. The method according to claim 17, wherein the predetermined
sTREM-1 value of step b) is the sTREM-1 median in the reference
population or the sTREM-1 third quartile in the reference
population.
21. The method according to claim 16, wherein the level of sTREM-1
measured at step a) is a level measured before the beginning of the
therapy.
22. The method according to claim 16, wherein the level of sTREM-1
measured at step a) is measured within the first 24 hours following
the diagnosis or the hospitalization of the human subject for an
inflammatory disorder.
23. The method according to claim 16, wherein the biological sample
is a blood sample, a serum sample or a plasma sample.
24. The method according to claim 16, wherein the level of sTREM-1
measured at step a) is a protein level.
25. The method according to claim 16, wherein the level of sTREM-1
measured at step a) is a protein level measured by enzyme-linked
immunosorbent assay (ELISA), electrochemiluminescence immunoassay
(ECLIA) or enzyme-linked fluorescent assay (ELFA).
26. The method according to claim 16, wherein the therapy is a
Triggering Receptors Expressed on Myeloid cells-1 (TREM-1)
inhibitor.
27. The method according to claim 26, wherein the TREM-1 inhibitor
is selected from the group consisting of peptides inhibiting the
function, activity or expression of TREM-1; antibodies directed to
TREM-1 and/or sTREM-1, or TREM-1 and/or sTREM-1 ligand; small
molecules inhibiting the function, activity or expression of
TREM-1; siRNAs directed to TREM-1; shRNAs directed to TREM-1;
antisense oligonucleotide directed to TREM-1; ribozymes directed to
TREM-1; and aptamers directed to TREM-1.
28. The method according to claim 27, wherein the TREM-1 inhibitor
is a peptide inhibiting the function, activity or expression of
TREM-1 by targeting TREM-1 ligand.
29. The method according to claim 27, wherein the TREM-1 inhibitor
is a peptide of 6 to 20 amino acids comprising an amino acid
sequence as set forth in SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10,
SEQ ID NO: 11, or SEQ ID NO: 12, or an amino acid sequence having
at least 90% identity with the amino acid sequence as set forth in
SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, or SEQ ID
NO: 12.
30. The method according to claim 27, wherein the TREM-1 inhibitor
is a peptide having an amino acid sequence as set forth in SEQ ID
NO: 9 or an amino acid sequence having at least 90% identity with
the amino acid sequence as set forth in SEQ ID NO: 9.
31. The method according to claim 16, wherein the inflammatory
disorder is selected from the group consisting of systemic
inflammatory response syndrome (SIRS), sepsis, and septic
shock.
32. The method according to claim 16, wherein the human subject is
suffering from an organ dysfunction defined as an acute change in
his/her sequential organ failure assessment (SOFA) score of at
least 2 points.
33. The method according to claim 31, wherein a human subject
suffering from SIRS, sepsis or septic shock susceptible to respond
to a therapy is a human subject susceptible to have his/her
sequential organ failure assessment (SOFA) score decrease following
the administration of the therapy.
34. The method according to claim 31, wherein a human subject
suffering from septic shock susceptible to respond to a therapy is
a human subject suffering from septic shock susceptible to reverse
a hypotensive shock within or after the administration period of
the therapy, wherein a hypotensive shock reversal is defined as the
absence of any vasopressor therapy during 24 hours.
35. An in vitro method for identifying a human subject suffering
from an inflammatory disorder susceptible to respond to a therapy,
said method comprising: a) measuring the level of soluble
Triggering Receptors Expressed on Myeloid cells-1 (sTREM-1) in a
biological sample from the human subject; b) comparing the level of
sTREM-1 measured at step a) to a predetermined sTREM-1 value; c)
identifying a human subject suffering from an inflammatory disorder
with a level of sTREM-1 measured at step a) higher than the
predetermined sTREM-1 value of step b) as susceptible to respond to
a therapy.
Description
FIELD
[0001] The present invention relates to the treatment of an
inflammatory disorder, preferably an acute inflammatory disorder,
in human subjects, in particular in a subpopulation of human
subjects susceptible to respond to a therapy, in particular to a
TREM-1 inhibitor. The present invention thus relates to methods for
identifying human subjects suffering from an inflammatory disorder,
preferably an acute inflammatory disorder such as systemic
inflammatory response syndrome (SIRS), sepsis or septic shock,
susceptible to respond to a therapy, in particular to a TREM-1
inhibitor.
BACKGROUND
[0002] Systemic inflammatory response syndrome (SIRS) is
characterized by systemic inflammation and widespread tissue
injury. SIRS may occur as a response to a nonspecific insult of
either infectious or non-infectious origin. Examples of insults of
non-infectious origin include, without being limited to, trauma,
thermal injury, pancreatitis, autoimmune disorders and surgery.
Examples of insults of infectious origin include bacterial
infections fungal infections and viral infections.
[0003] Sepsis is defined as a life-threatening organ dysfunction
caused by a dysregulated host response to infection (Singer et al.,
JAMA. 2016 Feb. 23; 315(8):801-10). Sepsis thus arises when the
body's response to an infection injures its own tissues and organs,
eventually leading to death in 30-50% of cases (Fleischmann et al.,
Dtsch Arztebl Int. 2016 Mar. 11; 113(10):159-66). Moreover, many
survivors suffer from post-sepsis syndrome, reporting an increase
in sensory, integumentary, digestive, breathing, chest pain, kidney
and musculoskeletal problems after sepsis (Huang et al., Int J Qual
Health Care. 2018 Jun. 19). According to the World Health
Organization, it is estimated that sepsis affects more than 30
million people worldwide every year, potentially leading to 6
million deaths.
[0004] Septic shock is defined as a subset of sepsis in which
particularly profound circulatory, cellular, and metabolic
abnormalities are associated with a greater risk of mortality than
with sepsis alone (Singer et al., JAMA. 2016 Feb. 23;
315(8):801-10). Clinically, patients with septic shock can be
identified as sepsis patients (i) suffering from hypotension
persisting after adequate fluid resuscitation requiring use of
vasopressors to maintain mean blood pressure of 65 mm Hg or greater
and (ii) having a serum lactate level greater than 2 mmol/L
(Shankar-Hari et al., JAMA. 2016 Feb. 23; 315(8):775-87).
[0005] Currently, there is no specific causal treatment for sepsis
or septic shock. Management of patients thus relies mainly on early
recognition allowing correct therapeutic measures to be started
rapidly, including administration of appropriate antibiotics,
source control measures when necessary, and resuscitation with
intravenous fluids and vasoactive drugs when needed (Cohen et al.,
Lancet Infect Dis. 2015 May; 15(5):581-614). Previous attempts to
develop treatments have failed, including therapies targeting
endotoxins and TLRs (Cuvier et al., Br J Clin Pharmacol. 2018 Jun.
8).
[0006] Recently, the Applicant developed a therapy which targets
the TREM-1 (triggering receptor expressed on myeloid cells-1)
pathway. TREM-1 is an immunoreceptor expressed by innate immune
cells (monocyte/macrophages, neutrophils, platelets, dendritic
cells) and endothelial cells. TREM-1 activation leads to cytokines
and chemokines production along with rapid neutrophil degranulation
and oxidative burst. TREM-1 function is to modulate/amplify, rather
than activate/initiate, inflammation by synergizing with pathogen
recognition receptors (PRRs), including Toll-like receptors (TLRs),
in order to trigger an exuberant immune response. Notably, the
TREM-1 pathway is involved in the pathophysiology of sepsis and
septic shock. The Applicant thus showed that TREM-1 inhibitors,
i.e., short TLT-1 (TREM-like transcript-1) peptides inhibiting
TREM-1 activity, may be used in the treatment of inflammatory
disorders, in particular acute inflammatory disorders such as
systemic inflammatory response syndrome (SIRS), sepsis or septic
shock (WO2011/124685). TLT-1 (Trem-Like Transcript-1) is a
receptor, member of the TREM family, exclusively expressed by
megakaryocytes and platelets.
[0007] However, for the TREM-1 inhibitor to provide the greatest
therapeutic benefit in the treatment of an acute inflammatory
disorder such as SIRS, sepsis or septic shock, there is still a
need to identify the human subjects suffering from an acute
inflammatory disorder susceptible to respond to a therapy, in
particular to a TREM-1 inhibitor. Notably, there is still a need to
identify the human subjects suffering from an acute inflammatory
disorder such as SIRS, sepsis or septic shock susceptible to
respond to a therapy, in particular to a TLT-1 peptide inhibiting
TREM-1 activity.
[0008] The Applicant now shows that measuring the level of soluble
TREM-1 (sTREM-1) in a biological sample from human subjects
suffering from septic shock and comparing their measured level of
sTREM-1 to a predetermined sTREM-1 value allows to identify the
human subjects suffering from septic shock susceptible to respond
to a TREM-1 inhibitor. In particular, the Applicant shows that
human subjects suffering from septic shock with a circulatory level
of sTREM-1 higher than the median sTREM-1 level predetermined in a
reference population of human subjects suffering from septic shock
are more likely to respond to, and thus benefit from, the
administration of a TLT-1 peptide inhibiting TREM-1 activity.
[0009] The present invention thus relates to an in vitro method for
identifying a human subject suffering from an inflammatory
disorder, preferably an acute inflammatory disorder such as SIRS,
sepsis or septic shock, susceptible to respond to a therapy, in
particular to a TREM-1 inhibitor, said method comprising: [0010] a)
measuring the level of soluble triggering receptors expressed on
myeloid cells-1 (sTREM-1) in a biological sample from the human
subject; [0011] b) comparing the level of sTREM-1 measured at step
a) to a predetermined sTREM-1 value; [0012] c) identifying a human
subject suffering from an inflammatory disorder, preferably an
acute inflammatory disorder such as SIRS, sepsis or septic shock
with a level of sTREM-1 measured at step a) higher than the
predetermined sTREM-1 value of step b) as susceptible to respond to
a therapy, in particular to a TREM-1 inhibitor.
[0013] The present invention also relates to a therapy, preferably
a TREM-1 inhibitor, for use in the treatment of an inflammatory
disorder, preferably an acute inflammatory disorder such as SIRS,
sepsis or septic shock, in a subject in need thereof, wherein a
level of sTREM-1 measured in a biological sample from the subject
and higher than a predetermined sTREM-1 value indicates that the
subject is susceptible to respond to the therapy, preferably to the
TREM-1 inhibitor.
SUMMARY
[0014] The present invention thus relates to an in vitro method for
identifying a human subject suffering from an inflammatory disorder
susceptible to respond to a therapy, preferably to a TREM-1
inhibitor, said method comprising: [0015] a) measuring the level of
soluble Triggering Receptors Expressed on Myeloid cells-1 (sTREM-1)
in a biological sample from the human subject; [0016] b) comparing
the level of sTREM-1 measured at step a) to a predetermined sTREM-1
value; [0017] c) identifying a human subject suffering from an
inflammatory disorder with a level of sTREM-1 measured at step a)
higher than the predetermined sTREM-1 value of step b) as
susceptible to respond to a therapy, preferably to a TREM-1
inhibitor.
[0018] In one embodiment, the predetermined sTREM-1 value of step
b) is obtained from a reference population. In one embodiment, the
reference population is a population of human patients suffering
from an inflammatory disorder, preferably systemic inflammatory
response syndrome (SIRS), sepsis or septic shock.
[0019] In one embodiment, the predetermined sTREM-1 value of step
b) is the sTREM-1 median in the reference population or the sTREM-1
third quartile in the reference population.
[0020] In one embodiment, the level of sTREM-1 measured at step a)
is a level measured before the beginning of the therapy, preferably
before the administration of a TREM-1 inhibitor.
[0021] In one embodiment, the level of sTREM-1 measured at step a)
is measured within the first 24 hours following the diagnosis or
the hospitalization of the human subject for an inflammatory
disorder.
[0022] In one embodiment, the biological sample is a blood sample,
a serum sample or a plasma sample.
[0023] In one embodiment, the level of sTREM-1 measured at step a)
is a protein level, preferably measured by ELISA,
electrochemiluminescence (ECL) also referred as
electrochemiluminescence immunoassay (ECLIA) or enzyme-linked
fluorescent assay (ELFA).
[0024] In one embodiment, the therapy comprises the administration
of a TREM-1 inhibitor, preferably selected from the group
consisting of peptides inhibiting the function, activity or
expression of TREM-1; antibodies directed to TREM-1 and/or sTREM-1,
or TREM-1 and/or sTREM-1 ligand; small molecules inhibiting the
function, activity or expression of TREM-1; siRNAs directed to
TREM-1; shRNAs directed to TREM-1; antisense oligonucleotide
directed to TREM-1; ribozymes directed to TREM-1 and aptamers
directed to TREM-1. In one embodiment, the therapy comprises the
administration of a TREM-1 inhibitor, said TREM-1 inhibitor being a
peptide targeting sTREM-1 ligand, preferably a peptide having an
amino acid sequence selected from the group consisting of SEQ ID
NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO:
12. In one embodiment, the therapy comprises the administration of
a TREM-1 inhibitor, said TREM-1 inhibitor being a peptide having an
amino acid sequence as set forth in SEQ ID NO: 9.
[0025] In one embodiment, the therapy is a TREM-1 inhibitor,
preferably selected from the group consisting of peptides
inhibiting the function, activity or expression of TREM-1;
antibodies directed to TREM-1 and/or sTREM-1, or TREM-1 and/or
sTREM-1 ligand; small molecules inhibiting the function, activity
or expression of TREM-1; siRNAs directed to TREM-1; shRNAs directed
to TREM-1; antisense oligonucleotide directed to TREM-1; ribozymes
directed to TREM-1 and aptamers directed to TREM-1. In one
embodiment, the TREM-1 inhibitor is a peptide inhibiting the
function, activity or expression of TREM-1 by targeting TREM-1
ligand, preferably a peptide having an amino acid sequence selected
from the group consisting of SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO:
10, SEQ ID NO: 11, and SEQ ID NO: 12. In one embodiment, the TREM-1
inhibitor is a peptide having an amino acid sequence as set forth
in SEQ ID NO: 9.
[0026] In one embodiment, the inflammatory disorder is selected
from the group comprising systemic inflammatory response syndrome
(SIRS), sepsis and septic shock, preferably the inflammatory
disorder is septic shock. In one embodiment, the human subject is
suffering from an organ dysfunction defined as an acute change in
his/her sequential organ failure assessment (SOFA) score of at
least 2 points.
[0027] In one embodiment, a human subject suffering from SIRS,
sepsis or septic shock susceptible to respond to a therapy,
preferably to a TREM-1 inhibitor, is a human subject susceptible to
have his/her sequential organ failure assessment (SOFA) score
decrease following the administration of the therapy, preferably
following the administration of a TREM-1 inhibitor. In one
embodiment, a human subject suffering from septic shock susceptible
to respond to a therapy, preferably to a TREM-1 inhibitor, is a
human subject suffering from septic shock susceptible to reverse a
hypotensive shock within or after the administration period of the
therapy, preferably of a TREM-1 inhibitor, wherein a hypotensive
shock reversal is defined as the absence of any vasopressor therapy
during 24 hours.
[0028] In the present invention, the following terms have the
following meanings: [0029] "About" preceding a figure encompasses
plus or minus 10%, or less, of the value of said figure. It is to
be understood that the value to which the term "about" refers is
itself also specifically, and preferably, disclosed. [0030] "APACHE
II" refers to "Acute Physiology And Chronic Health Evaluation II".
APACHE II is a scoring system commonly used to assess the severity
of disease in adult patients admitted in intensive care units and
determine their prognostic. APACHE II uses a point score ranging
from 0 to 71 that is based upon the initial values of 12 routine
physiologic measurements (i.e., variables), age, and previous
health status to provide a general measure of severity of disease
(Knaus et al., Crit Care Med. 1985 October; 13(10):818-29). [0031]
"APACHE III" refers to "Acute Physiology And Chronic Health
Evaluation III". APACHE III is a scoring system that was redefined
from the APACHE II scoring system in order to more accurately
predict hospital mortality risk for critically ill hospitalized
adults (Knaus et al., Chest. 1991 December; 100(6):1619-36). The
APACHE III scoring system is similar to the APACHE II scoring
system, except that several variables have been added to those used
in the APACHE II scoring system so that 17 variables are computed
into a point score ranging from 0 to 299. [0032] "APACHE IV" refers
to "Acute Physiology And Chronic Health Evaluation IV". APACHE IV
is an improved and updated model for estimating the risk of
short-term mortality as well as predicting the length of intensive
care unit (ICU) stay (Zimmerman et al., Crit Care Med. 2006 May;
34(5):1297-310). In the APACHE IV scoring system, a greater number
of variables are considered, notably mechanical ventilation,
thrombolysis, impact of sedation on Glasgow Coma Scale, rescaled
Glasgow Coma Scale, PaO.sub.2/FiO.sub.2 ratio and disease-specific
subgroups. APACHE IV uses a point score ranging from 0 to 286.
[0033] "Electrochemiluminescence immunoassay (ECLIA)" refers to an
immunoassay wherein the detection of the signal is based on
electrochemiluminescence, i.e., a form of chemiluminescence in
which the light-emitting chemiluminescent reaction is preceded by
an electrochemical reaction. [0034] "Fluid therapy" refers to a
therapy aiming at restoring and/or maintaining the volume and
composition of the body fluids to normal, in particular with
respect to the water-electrolyte balance. Fluid therapy thus aims
at correcting and/or preventing volume and/or electrolyte deficit.
[0035] "Immunodeficient" refers to a subject whose ability to
develop a normal immune response, for example to fight an
infectious disease or a cancer, is compromised or entirely absent.
Similarly, "immunocompromised" refers to a subject whose ability to
develop a normal immune response, for example to fight an
infectious disease or a cancer, is compromised "Immunosuppressed"
refers to a subject in whom the activation or efficacy of the
immune system is reduced, notably through the administration of an
immunosuppressive therapy. For example, immunosuppression in a
subject may be assessed through the measure of PD-1 level, the
measure of circulatory IL-7 level, or the measure of HLA-DR. [0036]
"Measuring" or "measurement", or alternatively "detecting" or
"detection", means assessing the presence, absence, quantity, or
amount (which can be an effective amount) of a given substance,
i.e., sTREM-1, within a biological sample from a human subject.
"Measuring" or "measurement", or alternatively "detecting" or
"detection" as used herein include the derivation of the
qualitative or quantitative concentration of said substance, i.e.,
sTREM-1, within the biological sample and within the human subject
(e.g., blood concentration or plasma concentration). [0037] "Organ
dysfunction score" or "organ dysfunction scoring system" as used
herein refers to a score used to assess organ dysfunction in a
human subject, notably in a human subject suffering from SIRS,
sepsis or septic shock, upon hospitalization, in particular upon
admission in ICU or emergency unit. Examples of organ dysfunction
scores include, without being limited to, the SOFA score, the qSOFA
score, the MODS (Multiple Organ Dysfunction Score), the P-MODS
(Pediatric Multiple Organ Dysfunction Score) and the LODS (Logistic
Organ Dysfunction System). [0038] "qSOFA score" refers to the quick
SOFA score (also known as quickSOFA). The qSOFA scoring system
relies on three criteria: respiratory rate .gtoreq.22 breaths/min;
altered mentation (Glasgow coma scale <15); and systolic blood
pressure .gtoreq.100 mm Hg (Seymour et al., JAMA. 2016 Feb. 23;
315(8):762-74). [0039] "Quantile(s)" refers to (a) cut-off value(s)
dividing the observations/measures made in a population into
equal-sized groups, each group comprising an equal percentage of
said observations/measures. As used herein, "quantile(s)" thus
refer to cut-off sTREM-1 value(s) dividing the sTREM-1 level
measured in a biological sample from each of the human subjects of
a reference population into equal-sized groups each comprising an
equal percentage of said measures of sTREM-1 level. In other words,
"quantile(s)" refers to cut-off sTREM-1 value(s) below or above
which lies a determined percentage of the sTREM-1 levels measured
in a reference population. For example, as used herein, "quartiles"
refer to the three cut-off sTREM-1 values dividing the sTREM-1
level measured in a reference population into four groups, each
comprising 25% of the sTREM-1 levels measured in the reference
population. It should be noted that "quantiles" may also sometimes
refer to the groups so defined by said cut-off value. For example,
"quartiles" may also sometimes refer to the four groups defined by
the three cut-off sTREM-1 values so dividing the sTREM-1 level
measured in a reference population. However, as used herein and
unless otherwise specified, the term "quantile" refers to a cut-off
value. [0040] "SAPS" refers to "Simplified Acute Physiology Score".
SAPS is a scoring system reflecting the risk of death in ICU
patients. SAPS relies on 14 biological and clinical variables (Le
Gall et al., Crit Care Med. 1984 November; 12(11):975-7). [0041]
"SAPS II" refers to "Simplified Acute Physiology Score II". SAPS II
is a scoring system for estimating in-hospital mortality in adult
patients admitted to the intensive care unit (ICU). SAPS II include
17 variables: 12 physiology variables, age, type of admission, and
3 variables regarding underlying diseases (Le Gall et al., JAMA.
1993 Dec. 22-29; 270(24):2957-63). SAPS II uses a point score
ranging from 0 to 163. [0042] "SAPS 3" refers to "Simplified Acute
Physiology Score III". SAPS 3 is a scoring system for predicting
hospital mortality of patients admitted to the intensive care unit
(ICU) (Metnitz et al., Intensive Care Med. 2005 October; 31(10):
1336-1344 and Moreno et al., Intensive Care Med. 2005 October;
31(10):1345-55). SAPS 3 is based on 20 different variables. [0043]
"Severity score" or "severity scoring system" as used herein refer
to a score used to assess the severity of the disease and/or the
prognosis of human subjects, notably of human subjects suffering
from SIRS, sepsis or septic shock, upon admission in ICU or
emergency unit. Examples of severity scores include, without being
limited to, the APACHE II score, the APACHE III score, the APACHE
IV score, the SAPS score, the SAPS II score and the SAPS 3 score.
[0044] "SIRS, sepsis or septic shock therapy" as used herein refers
to a therapy administered to a subject in need thereof for the
treatment of SIRS, sepsis or septic shock. In one embodiment, the
SIRS, sepsis or septic shock therapy is an immunomodulatory or an
anti-inflammatory therapy. Examples of immunomodulatory or an
anti-inflammatory therapies include, without being limited to,
checkpoint inhibitors such as anti-PD-1, anti-PD-L1 and anti-CTLA4;
TLR (Toll-like receptors) inhibitors; cytokine inhibitors such as
anti-cytokine or anti-cytokine receptors (for example IL-1RA for
interleukin-1 receptor antagonist); G-CSF (granulocyte-colony
stimulating factor); IL-7 (interleukin-7); inhibitors of
immunostimulants such as CD28 antagonist peptides and antibodies,
in particular monoclonal antibodies, against CD28; and cellular
therapies such as adoptive cell therapies. In one embodiment, the
SIRS, sepsis or septic shock therapy is an angiogenesis inhibitor,
in particular an angiopoietin-2 (Ang-2 or Ang2) inhibitor. In one
embodiment, the SIRS, sepsis or septic shock therapy is a TREM-1
inhibitor. In one embodiment, the TREM-1 inhibitor is a peptide
targeting TREM-1 ligand, preferably a peptide having an amino acid
sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID
NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12. [0045]
"Shock" or "hypotensive shock" or "distributive shock" refer to a
diminished or insufficient perfusion that impairs organ function
and is generally associated with decreased arterial blood pressure.
A hypotensive shock may be unresponsive to fluid resuscitation and
require a vasopressor therapy to increase the arterial blood
pressure, restore effective tissue perfusion and normalize cellular
metabolism. [0046] "Shock reversal" or "hypotensive shock reversal"
as used herein are defined as the absence of any vasopressor
therapy during at least 24 hours (i.e., not requiring to restart a
vasopressor therapy in the 24 hours following the end of a
vasopressor therapy). In one embodiment, shock reversal may occur
within the administration period of the SIRS, sepsis or septic
shock therapy. In one embodiment, shock reversal may occur 6, 12,
18, 24, 30, 36, 42, 48, 54, 60, 66 or 72 hours following the start
of the administration of the SIRS, sepsis or septic shock therapy.
In one embodiment, shock reversal may occur after the
administration period of the SIRS, sepsis or septic shock therapy.
In one embodiment, shock reversal may occur 6, 12, 18, 24, 30, 36,
42, 48, 54, 60, 66 or 72 hours following the end of the
administration of the SIRS, sepsis or septic shock therapy. [0047]
"SOFA score" refers to the Sequential Organ Failure Assessment
(SOFA) score (originally referred to as the Sepsis-related Organ
Failure Assessment) which is commonly used to assess the presence
of organ dysfunction. The SOFA scoring system (Vincent et al., Crit
Care Med. 1998 November; 26(11):1793-800) relies on the assessment
of the respiratory system (i.e., PaO2/FiO2 (mmHg)); of the nervous
system (i.e., Glasgow coma scale); of the cardiovascular system
(i.e., mean arterial pressure or administration of vasopressors
required); of the liver function (i.e., bilirubin (mg/dL or
.mu.mon)); of coagulation (i.e., platelet count); and of the kidney
function (i.e., creatinine (mg/dL or .mu.mon) or urine output
(mL/d)). [0048] "Standard of care" refers to the care routinely
provided to a patient suffering from an inflammatory disorder, in
particular systemic inflammatory response syndrome (SIRS), sepsis
or septic shock. In one embodiment, the patient is suffering from a
septic shock and the standard of care includes at least one of
antimicrobial therapy, fluid therapy, vasopressor therapy,
cardiovascular support, respiratory support, renal support, and
sedation. [0049] "TREM-1" refers to "triggering receptors expressed
on myeloid cells-1". TREM-1 is a membrane-bound glycoprotein
receptor belonging to the Ig superfamily that is notably expressed
on myeloid cells. TREM-1 activates downstream signaling pathways
with the help of an adapter protein called DAP12. TREM-1 comprises
three distinct domains: an Ig-like structure (mostly responsible
for ligand binding), a transmembrane part and a cytoplasmic tail
with associates with DAP12. Unless specified otherwise, the TREM-1
protein has an amino acid sequence as set forth in SEQ ID NO: 1,
corresponding to UniProtKB/Swiss-Prot accession number Q9NP99-1,
last modified on Oct. 1, 2000 and to UniProtKB accession number
Q38L15-1, last modified on Nov. 22, 2005. Several transcripts are
known for TREM-1. The transcript commonly referred to as TREM1-201
(transcript ID ensembl ENST00000244709.8) encodes an amino acid
sequence as set forth in SEQ ID NO: 1. The transcript commonly
referred to as TREM1-202, also known as TREM-1 isoform 2 (ensembl
transcript ID ENST00000334475.10) encodes an amino acid sequence as
set forth in SEQ ID NO: 2 (corresponding to UniProtKB/Swiss-Prot
accession number Q9NP99-2). The transcript commonly referred to as
TREM1-207, also known as TREM-1 isoform 3 (ensembl transcript ID
ENST00000591620.1) encodes an amino acid sequence as set forth in
SEQ ID NO: 3 (corresponding to UniProtKB/Swiss-Prot accession
number Q9NP99-3). The transcript commonly referred to as TREM1-204
(ensembl transcript ID ENST00000589614.5) encodes an amino acid
sequence as set forth in SEQ ID NO: 4 (corresponding to
UniProtKB/Swiss-Prot accession number K7EKM5-1, last modified Jan.
9, 2013). [0050] "sTREM-1", for "soluble triggering receptors
expressed on myeloid cells-1", refers to a soluble form of TREM-1
lacking the transmembrane and intracellular domains of TREM-1. In
one embodiment, sTREM-1 thus corresponds to the soluble form of the
extracellular domain of TREM-1. The soluble TREM-1 may be generated
by proteolytic cleavage of TREM-1 Ig-like ectodomain from the
membrane-anchored TREM-1 by matrix metalloproteinases (Gomez-Pina
et al., J Immunol. 2007 Sep. 15; 179(6):4065-73). In one
embodiment, sTREM-1 thus corresponds to a truncated TREM-1 shed
from the membrane of myeloid cells, in particular from activated
myeloid cells. It was also suggested that sTREM-1 results from an
alternative splicing of TREM-1 mRNA. A TREM-1 splice variant was
characterized in 2015 by Baruah et al. (J Immunol. 2015 Dec. 15;
195(12):5725-31), and was found to be secreted from primary and
secondary human neutrophil granules In one embodiment, sTREM-1 thus
corresponds to a TREM-1 splice variant, in particular to the TREM-1
transcript commonly referred to as TREM1-202, also known as TREM-1
isoform 2, encoding an amino acid sequence as set forth in SEQ ID
NO: 2. [0051] "sTREM-1 median" as used herein refers to a
predetermined sTREM-1 value obtained from a reference population
dividing the sTREM-1 levels measured in said reference population
into two groups, each comprising 50% of the sTREM-1 levels measured
in said reference population. [0052] "sTREM-1 terciles" as used
herein refers to predetermined sTREM-1 values obtained from a
reference population dividing the sTREM-1 levels measured in said
reference population into three groups, each comprising a third of
the sTREM-1 levels measured in said reference population. In one
embodiment, the sTREM-1 tercile is the last sTREM-1 tercile (i.e.,
the sTREM-1 second tercile), corresponding to the sTREM-1 value
below which two thirds of the sTREM-1 levels measured in the
reference population lie and above which one third of the sTREM-1
levels measured in the reference population lie.
[0053] "sTREM-1 quartiles" as used herein refers to predetermined
sTREM-1 values obtained from a reference population dividing the
sTREM-1 levels measured in said reference population into four
groups, each comprising 25% of the sTREM-1 levels measured in said
reference population. In one embodiment, the sTREM-1 quartile is
the last sTREM-1 quartile (i.e., the sTREM-1 third quartile also
referred to as Q3), corresponding to the sTREM-1 value below which
75% of the sTREM-1 levels measured in the reference population lie
and above which 25% of the sTREM-1 levels measured in the reference
population lie. [0054] "sTREM-1 quintiles" as used herein refers to
predetermined sTREM-1 values obtained from a reference population
dividing the sTREM-1 levels measured in said reference population
into five groups, each comprising 20% of the sTREM-1 levels
measured in said reference population. In one embodiment, the
sTREM-1 quintile is the last sTREM-1 quintile (i.e., the sTREM-1
fourth quintile), corresponding to the sTREM-1 value below which
80% of the sTREM-1 levels measured in the reference population lie
and above which 20% of the sTREM-1 levels measured in the reference
population lie. [0055] "sTREM-1 deciles" as used herein refers to
predetermined sTREM-1 values obtained from a reference population
dividing the sTREM-1 levels measured in said reference population
into ten groups, each comprising 10% of the sTREM-1 levels measured
in said reference population. In one embodiment, the sTREM-1 decile
is the last sTREM-1 decile (i.e., the sTREM-1 ninth decile),
corresponding to the sTREM-1 value below which 90% of the sTREM-1
levels measured in the reference population lie and above which 10%
of the sTREM-1 levels measured in the reference population lie.
[0056] "sTREM-1 percentiles" as used herein refers to a
predetermined sTREM-1 value obtained from a reference population
dividing the sTREM-1 levels measured in said reference population
into groups corresponding to a given percentage of the sTREM-1
levels measured in the reference population. As used herein, a
sTREM-1 percentile is thus a predetermined sTREM-1 value obtained
from a reference population below which a given percentage of the
sTREM-1 levels measured in said reference population lies. For
example, the 40th percentile is the sTREM-1 value obtained from a
reference population below which 40% of the sTREM-1 levels measured
in said reference population lie. [0057] "Therapeutically effective
amount" or "therapeutically effective dose" refer to the amount or
dose of therapy, preferably a TREM-1 inhibitor, that is aimed at,
without causing significant negative or adverse side effects to the
human subject, (1) delaying or preventing the onset of an
inflammatory disorder, preferably SIRS, sepsis or septic shock, in
the human subject; (2) reducing the severity or incidence of an
inflammatory disorder, preferably SIRS, sepsis or septic shock; (3)
slowing down or stopping the progression, aggravation, or
deterioration of one or more symptoms of an inflammatory disorder,
preferably SIRS, sepsis or septic shock, affecting the human
subject; (4) bringing about ameliorations of the symptoms of an
inflammatory disorder, preferably SIRS, sepsis or septic shock,
affecting the human subject; or (5) curing an inflammatory
disorder, preferably SIRS, sepsis or septic shock, affecting the
human subject. In one embodiment, the administration of a
therapeutically effective dose (or amount) of a therapy, preferably
of TREM-1 inhibitor, to a human subject suffering from an
inflammatory disorder, preferably SIRS, sepsis or septic shock,
aims at inducing in said subject at least one of the following:
[0058] reversal of a hypotensive shock, preferably within or after
the administration period of the therapy, preferably a TREM-1
inhibitor, for example over the 6, 12, 18 or 24 hours following the
end of said administration, wherein a hypotensive shock reversal is
defined as the absence of any vasopressor therapy during at least
24 hours (i.e., not requiring to restart a vasopressor therapy in
the 24 hours following the end of a vasopressor therapy); [0059] a
decrease of a severity score used to assess the severity of the
disease and/or the prognosis of subjects suffering from an
inflammatory disorder, preferably SIRS, sepsis or septic shock,
upon admission in ICU or emergency unit, such as the APACHE II
score, the APACHE III score, the APACHE IV score, the SAPS score,
the SAPS II score or the SAPS 3 score; [0060] a decrease of an
organ dysfunction score used to assess the presence of organ
dysfunction in a human subject suffering from an inflammatory
disorder, preferably SIRS, sepsis or septic shock, upon admission
in ICU or emergency unit, such as the SOFA score, the qSOFA score,
the MODS (Multiple Organ Dysfunction Score), the P-MODS (Pediatric
Multiple Organ Dysfunction Score) or the LODS (Logistic Organ
Dysfunction System); [0061] a decrease of the SOFA score and/or of
the qSOFA score, in particular over the 1, 2, 3, 4, 5, 6 or 7
day(s) following the start of the administration of the therapy,
preferably a TREM-1 inhibitor, preferably with reference to the
SOFA score and/or the qSOFA score assessed upon admission in ICU or
emergency unit or before the start of the administration of the
therapy; in one embodiment, said decrease of the SOFA score is a
decrease of at least 1 point (also referred as a delta of -1 point
or .DELTA.SOFA of -1 point), preferably of at least 1.5 point, in
particular at day 3 or at day 5 following the start of the
administration of the therapy, preferably with reference to the
SOFA score and/or the qSOFA score assessed upon admission in ICU or
emergency unit or before the start of the administration of the
therapy; [0062] decrease in the requirement for cardiovascular
support, for example a decrease in the use of vasopressor therapy,
in particular over the 1, 2, 3, 4, 5, 6 or 7 day(s) following the
start of the administration of the therapy, preferably a TREM-1
inhibitor, preferably with reference to the requirement for
cardiovascular support upon admission in ICU or emergency unit or
before the start of the administration of the therapy; [0063] a
decrease in the requirement for respiratory support, for example a
decrease in the use of invasive mechanical ventilation (IMV), in
particular over the 1, 2, 3, 4, 5, 6 or 7 day(s) following the
start of the administration of the therapy, preferably a TREM-1
inhibitor, preferably with reference to the requirement for
respiratory support upon admission in ICU or emergency unit or
before the start of the administration of the therapy; [0064] a
decrease in the requirement for renal support, for example a
decrease in the use of continuous or discontinuous renal
replacement therapy also referred to as RRT (e.g., dialysis), in
particular over the 1, 2, 3, 4, 5, 6 or 7 day(s) following the
start of the administration of the therapy, preferably a TREM-1
inhibitor, preferably with reference to the requirement for renal
support upon admission in ICU or emergency unit or before the start
of the administration of the therapy; [0065] a decrease in the risk
of reinfection, in particular a decrease in the risk of reinfection
in the 28, 90 or 365 days following the initial inflammatory
disorder, in particular the initial infection responsible for an
inflammatory disorder; [0066] an absence of reinfection, in
particular an absence of reinfection in the 28, 90 or 365 days
following the initial inflammatory disorder, in particular the
initial infection responsible for an inflammatory disorder; [0067]
a decrease in the risk of rehospitalization, in particular a
decrease in the risk of rehospitalization in the 28, 90 or 365 days
following the initial inflammatory disorder, in particular the
initial infection responsible for an inflammatory disorder; [0068]
an absence of rehospitalization, in particular an absence of
rehospitalization in the 28, 90 or 365 days following the first
hospitalization; [0069] an increase in the chance of survival, in
particular 1-year, 2-year, 3-year, 4-year, 5-year, 6-year, 7-year,
8-year, 9-year or 10-year survival, following the start of the
administration of the therapy, preferably a TREM-1 inhibitor; in
one embodiment the chance of survival, in particular in a subject
with multiple comorbidities, is assessed with the Charlson
Comorbidity Index (CCI) and an increase in the chance of 10-year
survival corresponds to a decrease of the Charlson Comorbidity
Index, preferably with reference to CCI assessed upon admission in
ICU or emergency unit or before the start of the administration of
the therapy; [0070] a decrease in the risk of sepsis-related death,
in particular at day 5, day 28, day 90 or day 365 following the
start of the administration of the therapy, preferably a TREM-1
inhibitor; [0071] a decrease in the risk of the all-cause death, in
particular at day 5, day 28, day 90 or day 365 following the start
of the administration of the therapy, preferably a TREM-1
inhibitor; [0072] a decrease in the risk of post-sepsis or
post-shock morbidity, in particular at day 5, day 28, day 90 or day
365 following the start of the administration of the therapy,
preferably a TREM-1 inhibitor; [0073] an increase in the quality of
life, in particular in the post-sepsis or post-shock quality of
life that may be assessed for example through an evaluation of
survival and quality-adjusted life years (QALYs), estimated from
the EuroQoL Quality of Life Scale commonly known as EQ5D. For
example, health-related quality of life (HRQoL) scores from the EQ
5D 5L (5-level EQ-5D version) may be calculated and converted to
utility scores, in particular at 3 months, 6 months, 9 months, 12
months, 18 months, 24 months or 36 months following the start of
the administration of the therapy. [0074] decrease in the level of
an inflammatory marker, such as, for example, CRP or IL6, IL-8,
IL-10, MCP-1 and TNF-.alpha.), in particular over the 1, 2, 3, 4,
5, 6 or 7 day(s) following the start of the administration of the
therapy, preferably a TREM-1 inhibitor, preferably with reference
to the level assessed upon admission in ICU or emergency unit or
before the start of the administration of the therapy; or [0075]
decrease in the level of an endothelial injury marker, such as, for
example, Ang-2, VCAM-1, VGEFR-1 and E-selectin, in particular over
the 1, 2, 3, 4, 5, 6 or 7 day(s) following the start of the
administration of the therapy, preferably a TREM-1 inhibitor,
preferably with reference to the level assessed upon admission in
ICU or emergency unit or before the start of the administration of
the therapy.
[0076] A therapeutically effective amount of a therapy, preferably
a TREM-1 inhibitor, may be administered for a prophylactic or
preventive action prior to the onset of an inflammatory disorder,
preferably SIRS, sepsis or septic shock; prior to the diagnosis of
an inflammatory disorder, preferably SIRS, sepsis or septic shock;
prior to the admission to ICU or emergency unit; or prior to the
start of vasopressor therapy. Alternatively, or additionally, a
therapeutically effective amount of a therapy, preferably a TREM-1
inhibitor, may be administered for a therapeutic action after the
onset of an inflammatory disorder, preferably SIRS, sepsis or
septic shock; after the diagnosis of an inflammatory disorder,
preferably SIRS, sepsis or septic shock; after the admission to ICU
or emergency unit; or after start of vasopressor therapy. [0077]
"TLT-1" refers to TREM-like transcript-1. TLT-1 is a receptor,
member of the TREM family, exclusively expressed by megakaryocytes
and platelets. TLT-1 contains a v-set Ig type-extracellular domain,
a transmembrane region and a cytoplasmic tail that comprises an
immunoreceptor tyrosine based inhibitory motif (ITIM) and a
polyproline-rich domain. [0078] "Treating" or "treatment" refers to
therapeutic treatment, to prophylactic or preventative measures, or
to both, wherein the object is to prevent, slow down (lessen) or
cure the targeted pathologic condition or disorder, i.e., an
inflammatory disorder, preferably SIRS, sepsis or septic shock. In
one embodiment of the present invention, "treating" or "treatment"
refers to a therapeutic treatment. In another embodiment of the
present invention, "treating" or "treatment" refers to a
prophylactic or preventive treatment. In yet another embodiment of
the present invention, "treating" or "treatment" refers to both a
prophylactic (or preventive) treatment and a therapeutic treatment.
Those in need of treatment include those already suffering from an
inflammatory disorder, preferably SIRS, sepsis or septic shock, as
well as those prone to develop an inflammatory disorder, preferably
SIRS, sepsis or septic shock, or those in whom an inflammatory
disorder, preferably SIRS, sepsis or septic shock is to be
prevented. A human subject suffering from an inflammatory disorder,
preferably SIRS, sepsis or septic shock, is successfully "treated"
if, after being administered a therapeutically effective amount of
a therapy, preferably a TREM-1 inhibitor, the human subject shows
at least one of the following: [0079] a reversal of a hypotensive
shock, preferably within or after the administration period of the
therapy, preferably a TREM-1 inhibitor, for example over the 6, 12,
18 or 24 hours following the end of said administration, wherein a
hypotensive shock reversal is defined as the absence of any
vasopressor therapy during at least 24 hours (i.e., not requiring
to restart a vasopressor therapy in the 24 hours following the end
of a vasopressor therapy); [0080] a decrease of a severity score
used to assess the severity of the disease and/or the prognosis of
subjects suffering from an inflammatory disorder, preferably SIRS,
sepsis or septic shock upon admission in ICU or emergency unit,
such as the APACHE II score, the APACHE III score, the APACHE IV
score, the SAPS II score or the SAPS 3 score; [0081] a decrease of
an organ dysfunction score used to assess the presence of organ
dysfunction in a human subject suffering from an inflammatory
disorder, preferably SIRS, sepsis or septic shock upon admission in
ICU or emergency unit, such as the SOFA score, the qSOFA score, the
MODS (Multiple Organ Dysfunction Score), the P-MODS (Pediatric
Multiple Organ Dysfunction Score) or the LODS (Logistic Organ
Dysfunction System); [0082] a decrease of the SOFA score and/or of
the qSOFA score, in particular over the 1, 2, 3, 4, 5, 6 or 7
day(s) following the start of the administration of therapy,
preferably a TREM-1 inhibitor, preferably with reference to the
SOFA score and/or the qSOFA score assessed upon admission in ICU or
emergency unit or before the start of the administration of the
therapy; in one embodiment, said decrease of the SOFA score is a
decrease of at least 1 point (also referred as a delta of -1 point
or .DELTA.SOFA of -1 point), preferably of at least 1.5 point, in
particular at day 3 or at day 5 following the start of the
administration of the therapy, preferably with reference to the
SOFA score and/or the qSOFA score assessed upon admission in ICU or
emergency unit or before the start of the administration of the
therapy; [0083] decrease in the requirement for cardiovascular
support, for example a decrease in the use of vasopressor therapy,
in particular over the 1, 2, 3, 4, 5, 6 or 7 day(s) following the
start of the administration of the therapy, preferably a TREM-1
inhibitor, preferably with reference to the requirement for
cardiovascular support upon admission in ICU or emergency unit or
before the start of the administration of the therapy; [0084] a
decrease in the requirement for respiratory support, for example a
decrease in the use of invasive mechanical ventilation (IMV), in
particular over the 1, 2, 3, 4, 5, 6 or 7 day(s) following the
start of the administration of the therapy, preferably a TREM-1
inhibitor, preferably with reference to the requirement for
respiratory support upon admission in ICU or emergency unit or
before the start of the administration of the therapy; [0085] a
decrease in the requirement for renal support, for example a
decrease in the use of continuous or discontinuous renal
replacement therapy also referred to as RRT (e.g., dialysis), in
particular over the 1, 2, 3, 4, 5, 6 or 7 day(s) following the
start of the administration of the therapy, preferably a TREM-1
inhibitor, preferably with reference to the requirement for renal
support upon admission in ICU or emergency unit or before the start
of the administration of the therapy; [0086] a decrease in the risk
of reinfection, in particular a decrease in the risk of reinfection
in the 28, 90 or 365 days following the initial inflammatory
disorder, in particular the initial infection responsible for an
inflammatory disorder; [0087] an absence of reinfection, in
particular an absence of reinfection in the 28, 90 or 365 days
following the initial inflammatory disorder, in particular the
initial infection responsible for an inflammatory disorder; [0088]
a decrease in the risk of rehospitalization, in particular a
decrease in the risk of reinfection in the 28, 90 or 365 days
following the initial inflammatory disorder, in particular the
initial infection responsible for an inflammatory disorder; [0089]
an absence of rehospitalization, in particular an absence of
rehospitalization in the 28, 90 or 365 days following the first
hospitalization; [0090] increase in the chance of survival, in
particular 1-year, 2-year, 3-year, 4-year, 5-year, 6-year, 7-year,
8-year, 9-year or 10-year survival, following the start of the
administration of the therapy, preferably a TREM-1 inhibitor; in
one embodiment the chance of survival, in particular in a subject
with multiple comorbidities, is assessed with the Charlson
Comorbidity Index (CCI) and an increase in the chance of 10-year
survival corresponds to a decrease of the Charlson Comorbidity
Index, preferably with reference to CCI assessed upon admission in
ICU or emergency unit or before the start of the administration of
the therapy; [0091] a decrease in the risk of sepsis-related death,
in particular at day 5, day 28, day 90 or day 365 following the
start of the administration of the therapy, preferably a TREM-1
inhibitor; [0092] a decrease in the risk of the all-cause death, in
particular at day 5, day 28, day 90 or day 365 following the start
of the administration of the therapy, preferably a TREM-1
inhibitor; [0093] a decrease in the risk of post-sepsis or
post-shock morbidity, in particular at day 5, day 28, day 90 or day
365 following the start of the administration of the therapy,
preferably a TREM-1 inhibitor; [0094] an increase in the quality of
life, in particular in the post-sepsis or post-shock quality of
life, that may be assessed for example through an evaluation of
survival and quality-adjusted life years (QALYs), estimated from
the EQ5D. For example, health-related quality of life (HRQoL)
scores from the EQ 5D 5L may be calculated and converted to utility
scores, in particular at 3 months, 6 months, 9 months, 12 months,
18 months, 24 months or 36 months following the start of the
administration of the therapy; [0095] a decrease in the level of an
inflammatory marker, such as, for example, CRP or IL6, IL-8, IL-10,
MCP-1 and TNF-.alpha.), in particular over the 1, 2, 3, 4, 5, 6 or
7 day(s) following the start of the administration of the therapy,
preferably a TREM-1 inhibitor, preferably with reference to the
level assessed upon admission in ICU or emergency unit or before
the start of the administration of the therapy; or [0096] a
decrease in the level of an endothelial injury marker, such as, for
example, Ang-2, VCAM-1, VGEFR-1 and E-selectin, in particular over
the 1, 2, 3, 4, 5, 6 or 7 day(s) following the start of the
administration of the therapy, preferably a TREM-1 inhibitor,
preferably with reference to the level assessed upon admission in
ICU or emergency unit or before the start of the administration of
the therapy.
[0097] The above parameters for assessing successful treatment and
improvement in the disease are readily measurable by routine
procedures familiar to a physician. [0098] "TREM-1 inhibitor"
refers to an active agent able to inhibit TREM-1 function, activity
or expression. Example of TREM-1 inhibitors include, without begin
limited to, peptides inhibiting the function, activity or
expression of TREM-1; antibodies directed to TREM-1 and/or sTREM-1,
or TREM-1 and/or sTREM-1 ligand; small molecules inhibiting the
function, activity or expression of TREM-1; siRNAs directed to
TREM-1; shRNAs directed to TREM-1; antisense oligonucleotide
directed to TREM-1; ribozymes directed to TREM-1 and aptamers
directed to TREM-1. [0099] "Vasopressor therapy" refers to a
therapy used in the treatment of shock, i.e., hypotensive shock,
defined as a diminished or insufficient perfusion that impairs
organ function and is generally associated with decreased arterial
blood pressure. In particular, vasopressor therapy is required when
the hypotension is unresponsive to fluid resuscitation. Vasopressor
therapy aims at increasing the arterial blood pressure and
maintaining an adequate blood pressure, restoring effective tissue
perfusion and normalizing cellular metabolism. Examples of
vasopressor therapies include, without limitation, the
administration of vasoactive catecholamine hormones such as
norepinephrine, dopamine, epinephrine; vasopressin; and/or
phenylephrine.
BRIEF DESCRIPTION OF THE DRAWINGS
[0100] FIGS. 1A-1C show that sTREM-1 release depends on TREM-1
dimerization. 1A is a graph showing TREM-1 expression by flow
cytometry on U937 (grey) and U937-vitD cells (dark-grey) compared
to isotype control (light-grey). 1B is a set of photographs showing
TREM-1 expression and cell nuclei staining by confocal microscopy.
White arrows indicate the clustering and dimerization of TREM-1 at
the membrane of U937-vitD cells after incubation with LPS. 1C is a
graph showing sTREM-1 concentration in the supernatant of U937 and
U937-vitD cells after 30 minutes incubation in resting conditions
or with APMA, Pro-MMP9, Act-MMP9 or LPS (100 ng/mL as indicated.
LOQ: Limit of Quantification.
[0101] FIGS. 2A-2B are a set of graphs showing that TREM-1 at
baseline is correlated to 28-day mortality. ROC plot (2A) and
Kaplan-Meier plot (2B) by quartiles for 28-day mortality. Mortality
rates (at day 28) per quartile are 12%, 30%, 37% and 49% for Q1-Q4,
respectively. Overall mortality rate is 32%. Optimal cut point was
determined to be at 408 pg/mL.
[0102] FIGS. 3A-3B are a set of boxplots of sTREM-1 at baseline
(day 1) for 28-day mortality (3A) and 90-day mortality (3B).
Chi.sup.2 of Kruskal-Wallis test is 28.8 and 29.5, respectively
(both p<0.0001).
[0103] FIGS. 4A-4B are a set of boxplots of sTREM-1 at day 2 for
28-day mortality (4A) and 90-day mortality (4B). Chi.sup.2 of
Kruskal-Wallis test is 30.2 and 30.8, respectively (both
p<0.0001).
[0104] FIGS. 5A-5B are a set of boxplots of sTREM-1 at day 3 for
28-day mortality (5A) and 90-day mortality (5B). Chi.sup.2 of
Kruskal-Wallis test is 16.9 (p=0.0001) and 19.1 (p=0.0002).
[0105] FIGS. 6A-6C are a set of boxplots of sTREM-1 at day 1 (6A),
day 2 (6B), day 3 (6C) for renal replacement therapy (RRT) at
admission (RRT adm) and during ICU stay (RRT later). Chi.sup.2 of
Kruskal-Wallis test is 54.7 (p<0.0001) and 68.7 (p<0.0001)
and 44 (p<0.0001) respectively.
[0106] FIG. 7 is a scheme depicting the two stages of the phase IIa
clinical trial assessing the administration of nangibotide (also
known as motrem) to septic shock patients (NCT03158948).
[0107] FIG. 8 is a scheme depicting patient randomization in each
treatment group.
[0108] FIGS. 9A-9B are a set of graphs depicting the
pharmacokinetics of nangibotide. 9A depicts nangibotide kinetics in
each group (i.e., patients who received 0.3 mg/kg/h, patients who
received 1.0 mg/kg/h and patients who received 3.0 mg/kg/h) from
the start of the infusion (day 0) to the end of infusion (day 5)
and 9B depicts nangibotide kinetics in each group (i.e., patients
who received 0.3 mg/kg/h, patients who received 1.0 mg/kg/h and
patients who received 3.0 mg/kg/h) after the end of the infusion.
Circles: 0.3 mg/kg/h; squares: 1 mg/kg/h; triangles: 3 mg/kg/h.
[0109] FIGS. 10A-10C are a set of graphs depicting the SOFA
evolution over time change versus baseline (LOCF) in the whole
population (10A), patients with low sTREM-1 at baseline (below the
median 433 pg/mL) (10B), patients with high sTREM-1 at baseline
(above the median 433 pg/mL) (10C).
[0110] FIGS. 11A-11C are a set of graphs depicting organ support
free days in patients with high levels of sTREM-1 at baseline
(above the median 433 pg/mL): vasopressor free days alive (11A),
invasive mechanical ventilation (IMV) free days alive (11B), renal
failure and continuous renal replacement therapy (CRRT) free days
alive (11C).
[0111] FIGS. 12A-12B are a set of graphs depicting Ang-2
concentration change from baseline at day 3 (12A) and day 5 (12B),
in "all" (corresponding to the whole population), in the "G1"
subgroup (corresponding to patients with low sTREM-1 at baseline,
i.e., below the median 433 pg/mL), and "the G2" subgroup
(corresponding to patients with high sTREM-1 at baseline, i.e.,
above the median 433 pg/mL).
[0112] FIGS. 13A-13B are a set of graphs depicting IL-6
concentration change from baseline at Day 3 (13A) and Day 5 (13B),
in "all" (corresponding to the whole population), in the "G1"
subgroup (corresponding to patients with low sTREM-1 at baseline,
i.e., below the median 433 pg/mL), and "the G2" subgroup
(corresponding to patients with high sTREM-1 at baseline, i.e.,
above the median 433 pg/mL).
[0113] FIGS. 14A-14B are a set of graphs. 14A depicts the sTREM-1
change at day 5/EOI (end of infusion) from baseline in all
population, in the subgroup G1 (patients with sTREM-1 baseline
below the median) and in the subgroup G2 (patients with sTREM-1
baseline above the median). 14B depicts the association between
delta SOFA at day 5 (delta between day 5/EOI and baseline).
DETAILED DESCRIPTION
[0114] The present invention relates to an in vitro method for
identifying a human subject suffering from an inflammatory disorder
susceptible to respond to a therapy, said method comprising: [0115]
a) measuring the level of soluble triggering receptors expressed on
myeloid cells-1 (sTREM-1) in a biological sample from the human
subject; [0116] b) comparing the level of sTREM-1 measured at step
a) to a predetermined sTREM-1 value; [0117] c) identifying a human
subject suffering from an inflammatory disorder with a level of
sTREM-1 measured at step a) higher than the predetermined sTREM-1
value of step b) as susceptible to respond to a therapy.
[0118] In other words, the present invention relates to an in vitro
method for identifying a human subject suffering from an
inflammatory disorder susceptible to respond to a therapy, wherein
a level of soluble Triggering Receptors Expressed on Myeloid
cells-1 (sTREM-1) measured in a biological sample from the human
subject higher than a predetermined sTREM-1 value indicates that
the human subject is susceptible to respond to a therapy.
[0119] The present invention also relates to an in vitro method for
selecting a human subject suffering from an inflammatory disorder,
for treatment with a therapy, preferably a TREM-1 inhibitor, said
method comprising: [0120] a) measuring the level of soluble
triggering receptors expressed on myeloid cells-1 (sTREM-1) in a
biological sample from the human subject; [0121] b) comparing the
level of sTREM-1 measured at step a) to a predetermined sTREM-1
value; [0122] c) selecting a human subject suffering from an
inflammatory disorder with a level of sTREM-1 measured at step a)
higher than the predetermined sTREM-1 value of step b) for
treatment with a therapy, preferably a TREM-1 inhibitor.
[0123] Another object of the invention is a therapy, preferably a
TREM-1 inhibitor, for use in the treatment of an inflammatory
disorder in a subject in need thereof, wherein a level of sTREM-1
measured in a biological sample from the subject and higher than a
predetermined sTREM-1 value indicates that the subject is
susceptible to respond to the therapy, preferably to the TREM-1
inhibitor.
[0124] Thus, the present invention also relates to a method for
treating an inflammatory disorder in a human subject identified as
susceptible to respond to a therapy, preferably to a TREM-1
inhibitor, said method comprising:
identifying a human subject suffering from an inflammatory disorder
susceptible to respond to a therapy, preferably to a TREM-1
inhibitor by: [0125] a) obtaining a biological sample from the
human subject and measuring the level of soluble triggering
receptors expressed on myeloid cells-1 (sTREM-1) in said biological
sample; [0126] b) comparing the level of sTREM-1 measured at step
a) to a predetermined sTREM-1 value; and [0127] c) identifying a
human subject suffering from an inflammatory disorder with a level
of sTREM-1 measured at step a) higher than the predetermined
sTREM-1 value of step b) as susceptible to respond to a therapy,
preferably to a TREM-1 inhibitor; administering a therapy,
preferably a TREM-1 inhibitor, to the subject suffering from an
inflammatory disorder identified as susceptible to respond to a
therapy, preferably to a TREM-1 inhibitor, thereby treating an
inflammatory disorder in a human subject identified as susceptible
to respond to a therapy, preferably to a TREM-1 inhibitor.
[0128] TREM-1 (triggering receptor expressed on myeloid cells-1) is
a glycoprotein receptor belonging to the Ig superfamily that is
expressed notably on myeloid cells. sTREM-1 is a soluble form of
TREM-1 lacking the transmembrane and intracellular domains of
TREM-1. Without wishing to be bound to a theory, the Applicant
suggest that PRRs (Pathogen Recognition Receptors) engagement,
including Nod-like receptors (NLRs) and Toll-like receptors (TLRs),
induces the upregulation of TREM-1 expression and/or its
mobilization and clustering at the cell membrane, which lead to its
dimerization and multimerization. Said NLRs and TLRs activation can
occur by linking DAMPs (Danger Associated Molecular Patterns) or
PAMPs (Pathogen Associated Molecular Patterns). In particular said
NLRs and TLRs activation can occur under sterile inflammatory
conditions by linking DAMPs (Danger Associated Molecular Patterns)
and/or alarmins, or under infectious conditions by linking PAMPs
(Pathogen Associated Molecular Patterns). This activation of NLRs
and TLRs induces the upregulation of proteases, in particular of
metalloproteinases, which in turn, among a number of targets, will
induce the liberation of a soluble TREM-1 through proteolytic
cleavage of membrane-anchored TREM-1 (Gomez-Pina et al., J Immunol.
2007 Sep. 15; 179(6):4065-73). Said proteolytic cleavage depends on
the dimerization of the TREM-1 receptor. sTREM-1 is thus shed from
the membrane of myeloid cells, in particular from activated myeloid
cells and sTREM-1 release is a marker of TREM-1 activation. In one
embodiment, sTREM-1 corresponds to the soluble form of the
extracellular domain of TREM-1. In one embodiment, sTREM-1
corresponds to a truncated TREM-1 shed from the membrane of myeloid
cells, in particular from activated myeloid cells.
[0129] sTREM-1 may also results from an alternative splicing of
TREM-1 mRNA. A TREM-1 splice variant was characterized in 2015 by
Baruah et al., (J Immunol. 2015 Dec. 15; 195(12):5725-31) and was
found to be secreted from primary and secondary human neutrophil
granules. In one embodiment, sTREM-1 corresponds to a TREM-1 splice
variant. In one embodiment, sTREM-1 corresponds to the TREM-1
transcript commonly referred to as TREM1-202, also known as TREM-1
isoform 2, encoding an amino acid sequence as set forth in SEQ ID
NO: 2. In one embodiment, sTREM-1 thus has an amino acid sequence
as set forth in SEQ ID NO: 2 (MRKTRLWGLLWMLFVSELRAAT
KLTEEKYELKEGQTLDVKCDYTLEKFASSQKAWQIIRDGEMPKTLACTERPSK
NSHPVQVGRIILEDYHDHGLLRVRMVNLQVEDSGLYQCVIYQPPKEPHMLFDRI
RLVVTKGFRCSTLSFSWLVDS)
[0130] In one embodiment, sTREM-1 has an amino acid sequence as set
forth in SEQ ID NO: 5
(ATKLTEEKYELKEGQTLDVKCDYTLEKFASSQKAWQIIRDGEMPKTLACTERP
SKNSHPVQVGRIILEDYHDHGLLRVRMVNLQVEDSGLYQCVIYQPPKEPHMLF
DRIRLVVTKGFSGTPGSNENSTQNVYKIPPTTTKALCPLYTSPRTVTQAPPKSTA
DVSTPDSEINLTNVTDIIRVPVFN), corresponding to amino acids 21 to 205
of SEQ ID NO: 1.
[0131] In another embodiment, sTREM-1 has an amino acid sequence as
set forth in SEQ ID NO: 6
(LKEGQTLDVKCDYTLEKFASSQKAWQIIRDGEMPKTLACTERPSKNSH
PVQVGRIILEDYHDHGLLRVRMVNLQVEDSGLYQCVIYQPPKEPHMLFDRIRLV
VTKGFSGTPGSNENSTQNVYKIPPTTTKALCPLYTSPRTVTQAPPKSTADVSTPD
SEINLTNVTDIIRVPVFN), corresponding to amino acids 31 to 205 of SEQ
ID NO: 1.
[0132] In another embodiment, sTREM-1 comprises an amino sequence
as set forth in SEQ ID NO: 19
(LKEGQTLDVKCDYTLEKFASSQKAWQIIRDGEMPKTLACTERPSKNSH
PVQVGRIILEDYHDHGLLRVRMVNLQVEDSGLYQCVIYQPPKEPHMLFDRIRLV VTKGF),
corresponding to amino acids 31 to 137 of SEQ ID NO: 1, and has a
length of 200 amino acids or less, preferably of 185 amino acids or
less.
[0133] In another embodiment, sTREM-1 has an amino acid sequence as
set forth in SEQ ID NO: 2, SEQ ID NO: 5 or SEQ ID NO: 6. In another
embodiment, sTREM-1 has an amino acid sequence as set forth in SEQ
ID NO: 5 or in SEQ ID NO: 6.
[0134] In one embodiment, sTREM-1 is a variant of SEQ ID NO: 2, a
variant of SEQ ID NO: 5 or a variant of SEQ ID NO: 6. In one
embodiment, sTREM-1 is a variant of SEQ ID NO: 5 or a variant of
SEQ ID NO: 6.
[0135] In one embodiment, a variant of SEQ ID NO: 2, SEQ ID NO: 5
or SEQ ID NO: 6 is an amino acid sequence comprising at least 25
contiguous amino acids, preferably at least 50, 60, 70, 80, 90,
100, 110, 120, 130, 140, 150, 160, 165, 170, 175, 180 or 185
contiguous amino acids of the amino acid sequence of SEQ ID NO: 2,
SEQ ID NO: 5 or SEQ ID NO: 6, respectively.
[0136] In another embodiment, a variant of SEQ ID NO: 2, SEQ ID NO:
5 or SEQ ID NO: 6 is an amino acid sequence comprising the amino
acid sequence of SEQ ID NO: 2, SEQ ID NO: 5 or SEQ ID NO: 6,
respectively, and additional amino acids in C-term or in N-term of
SEQ ID NO: 2, SEQ ID NO: 5 or SEQ ID NO: 6, wherein the number of
additional amino acids ranges from 1 to 50, preferably from 1 to
20, more preferably from 1 to 10 amino acids, such as, for example,
1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids in C-term and/or 1, 2,
3, 4, 5, 6, 7, 8, 9 or 10 amino acids in N-term.
[0137] In another embodiment, a variant of SEQ ID NO: 2, SEQ ID NO:
5 or SEQ ID NO: 6 is an amino acid sequence that typically differs
from the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 5 or SEQ
ID NO: 6 through one or more amino acid substitution(s),
deletion(s), addition(s) and/or insertion(s). In one embodiment,
said substitution(s), deletion(s), addition(s) and/or insertion(s)
may affect 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids.
[0138] In another embodiment, a variant of SEQ ID NO: 2, SEQ ID NO:
5 or SEQ ID NO: 6 is an amino acid sequence of at least 25 amino
acids, preferably of at least 50, 60, 70, 80, 90, 100, 110, 120,
130, 140, 150, 160, 165, 170, 175, 180 or 185 amino acids having at
least 60%, 65%, 70%, 75%, 80%, 90%, 95%, or at least 96%, 97%, 98%,
99% or more identity with the amino acid sequence of SEQ ID NO: 2,
SEQ ID NO: 5 or SEQ ID NO: 6, respectively.
[0139] The term "identity" or "identical", when used in a
relationship between the sequences of two or more polypeptides,
refers to the degree of sequence relatedness between polypeptides,
as determined by the number of matches between strings of two or
more amino acid residues. "Identity" measures the percent of
identical matches between the smaller of two or more sequences with
gap alignments (if any) addressed by a particular mathematical
model or computer program (i.e., "algorithms"). Identity of related
polypeptides can be readily calculated by known methods. Such
methods include, but are not limited to, those described in
Computational Molecular Biology, Lesk, A. M., ed., Oxford
University Press, New York, 1988; Biocomputing: Informatics and
Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993;
Computer Analysis of Sequence Data, Part 1, Griffin, A. M., and
Griffin, H. G., eds., Humana Press, New Jersey, 1994; Sequence
Analysis in Molecular Biology, von Heinje, G., Academic Press,
1987; Sequence Analysis Primer, Gribskov, M. and Devereux, J.,
eds., M. Stockton Press, New York, 1991; and Carillo et al., SIAM
J. Applied Math. 48, 1073 (1988). Preferred methods for determining
identity are designed to give the largest match between the
sequences tested. Methods of determining identity are described in
publicly available computer programs. Preferred computer program
methods for determining identity between two sequences include the
GCG program package, including GAP (Devereux et al., Nucl. Acid.
Res. \2, 387 (1984); Genetics Computer Group, University of
Wisconsin, Madison, Wis.), BLASTP, BLASTN, and FASTA (Altschul et
al., J. MoI. Biol. 215, 403-410 (1990)). The BLASTX program is
publicly available from the National Center for Biotechnology
Information (NCBI) and other sources (BLAST Manual, Altschul et al.
NCB/NLM/NIH Bethesda, Md. 20894; Altschul et al., supra). The
well-known Smith Waterman algorithm may also be used to determine
identity.
[0140] In one embodiment, the variant of SEQ ID NO: 2, SEQ ID NO: 5
or SEQ ID NO: 6 is not SEQ ID NO: 1.
[0141] In one embodiment, sTREM-1 is a fragment of SEQ ID NO: 2, a
fragment of SEQ ID NO: 5 or a fragment of SEQ ID NO: 6. In one
embodiment, sTREM-1 is a fragment of SEQ ID NO: 5 or a fragment of
SEQ ID NO: 6.
[0142] In one embodiment, a fragment of SEQ ID NO: 2, SEQ ID NO: 5
or SEQ ID NO: 6 is an amino acid sequence comprising at least 25
contiguous amino acids, preferably of at least 50, 60, 70, 80, 90,
100, 110, 120, 130, 140, 150, 160, 165, 170, 175, 180 or 185
contiguous amino acids of the amino acid sequence of SEQ ID NO: 2,
SEQ ID NO: 5 or SEQ ID NO: 6, respectively.
[0143] In one embodiment, sTREM-1 corresponds to the extracellular
fragment generated by cleavage of the membrane-bound TREM-1 having
an amino acid sequence as set forth in SEQ ID NO: 1 by a protease,
preferably a matrix metallopeptidase, more preferably by the matrix
metalloproteinase 9 (MMP9).
[0144] As used herein, "biological sample" refers to a biological
sample isolated from a human subject and can include, by way of
example and not limitation, bodily fluids, cell samples and/or
tissue extracts such as homogenates or solubilized tissue obtained
from a human subject.
[0145] In one embodiment, the method of the invention does not
comprise obtaining a biological sample from a subject. In one
embodiment, the biological sample from the human subject is a
biological sample previously obtained from the human subject. Said
biological sample may be conserved in adequate conditions before
being used in the method of the invention.
[0146] In one embodiment, the biological sample from the human
subject is a body fluid sample. Examples of body fluids include,
without being limited to, blood, plasma, serum, lymph, urine,
bronchioalveolar lavage fluid, cerebrospinal fluid, sweat or any
other bodily secretion or derivative thereof.
[0147] As used herein, "blood" includes whole blood, plasma, serum,
circulating epithelial cells, constituents, or any other derivative
of blood.
[0148] In one embodiment, the biological sample from the human
subject is a blood sample. In one embodiment, the biological sample
from the human subject is a whole blood sample or a plasma sample.
Methods for obtaining a plasma sample are routinely used in
clinical laboratories. In one embodiment, the whole blood sample or
the plasma sample from the human subject is processed to obtain a
serum sample. Methods for obtaining a serum sample from a whole
blood sample or a plasma sample are routinely used in clinical
laboratories.
[0149] In another embodiment, the biological sample from the human
subject is a tissue extract. Tissue extracts are obtained routinely
from tissue biopsy and autopsy material.
[0150] According to the present invention, the term "level" as used
herein refers to the expression level of sTREM-1. It can refer
alternatively to the transcription level of sTREM-1 or to the
translation level of sTREM-1. The expression level may be detected
intracellularly or extracellularly.
[0151] The level of TREM-1 may be measured with a point-of-care
testing (POCT) or bedside testing; with a near-to-patient testing;
or with a central laboratory assay.
[0152] Methods for measuring the expression level such as a
transcription level or a translation level are well-known to the
skilled artisan and include, but are not limited to, RT-PCR,
RT-qPCR, Northern Blot, hybridization techniques such as, for
example, use of microarrays, and combination thereof including but
not limited to, hybridization of amplicons obtained by RT-PCR,
sequencing such as, for example, next-generation DNA sequencing
(NGS) or RNA-seq (also known as "Whole Transcriptome Shotgun
Sequencing") and the like, immunohistochemistry, Multiplex methods
(such as Luminex), western blot, enzyme-linked immunosorbent assay
(ELISA), sandwich ELISA, multiplex ELISA, electrochemiluminescence
(ECL) also referred as electrochemiluminescence immunoassay (ECLIA)
(such as Elecsys.RTM., Roche Diagnostics), enzyme-linked
fluorescent assay (ELFA) (such as VIDAS.RTM., Biomerieux),
fluorescent-linked immunosorbent assay (FLISA), enzyme immunoassay
(EIA), radioimmunoassay (RIA), flow cytometry (FACS), surface
plasmon resonance (SPR), biolayer interferometry (BLI),
immunochromatographic assay (ICA) (such as NEXUS IB10, Sphingotech)
and mass spectrometry-based approaches.
[0153] According to one embodiment, the term "level" as used herein
refers to the quantity, amount or concentration of sTREM-1. Thus,
the level of sTREM-1 measured in a biological sample from a human
subject refers to the quantity, amount or concentration of sTREM-1
in said biological sample.
[0154] According to one embodiment, the level of sTREM-1 refers to
a protein level, a protein quantity, a protein amount or a protein
concentration.
[0155] In one embodiment, the level of sTREM-1 refers to the level
of the amino acid sequences as set forth in SEQ ID NO: 2, SEQ ID
NO: 5 and/or SEQ ID NO: 6, and/or fragments and/or variants thereof
as described hereinabove. In one embodiment, the level of sTREM-1
refers to the level of the amino acid sequences as set forth in SEQ
ID NO: 5 and/or SEQ ID NO: 6, and/or fragments and/or variants
thereof as described hereinabove.
[0156] In one embodiment, the biological sample from a human
subject is a blood sample and the level of sTREM-1 measured in said
biological sample corresponds to the blood concentration of sTREM-1
of said human subject.
[0157] In one embodiment, the biological sample from a human
subject is a plasma sample and the level of sTREM-1 measured in
said biological sample corresponds to the plasma concentration of
sTREM-1 of said human subject.
[0158] In one embodiment, the biological sample from a human
subject is a serum sample and the level of sTREM-1 measured in said
biological sample corresponds to the serum concentration of sTREM-1
of said human subject.
[0159] According to the invention, the level of sTREM-1 may be
measured by any known method in the art.
[0160] Methods for measuring the level of sTREM-1, in particular a
sTREM-1 protein level, in a biological sample as described
hereinabove are well-known to the skilled artisan and include,
without being limited to, immunohistochemistry, Multiplex methods
(such as Luminex), western blot, enzyme-linked immunosorbent assay
(ELISA), sandwich ELISA, multiplex ELISA, electrochemiluminescence
immunoassay (ECLIA) (such as Elecsys.RTM., Roche Diagnostics),
enzyme-linked fluorescent assay (ELFA) (such as VIDAS.RTM.,
Biomerieux), fluorescent-linked immunosorbent assay (FLISA), enzyme
immunoassay (EIA), radioimmunoassay (RIA), flow cytometry (FACS)
surface plasmon resonance (SPR), biolayer interferometry (BLI),
immunochromatographic assay (ICA) (such as NEXUS IB10, Sphingotech)
and mass spectrometry-based approaches.
[0161] Typically, measuring the level of sTREM-1 in a biological
sample as described hereinabove may comprise contacting the
biological sample with a binding partner capable of selectively
interacting with sTREM-1 in the biological sample. In one
embodiment, the binding partner is an antibody, such as, for
example, a monoclonal antibody or an aptamer.
[0162] In one embodiment, measuring the level of sTREM-1 in a
biological sample as described hereinabove comprises the use of an
antibody, such as a polyclonal or monoclonal antibody.
[0163] Examples of antibodies allowing the detection of sTREM-1
include, without being limited to, the polyclonal antibody raised
against Met1-Arg200 amino acids of human TREM-1 (reference AF1278
from R&D Systems), the monoclonal antibody raised against
Ala21-Asn205 of human TREM-1 (reference MAB1278 from R&D
Systems), the purified anti-human CD354 (TREM-1) antibody (clone
TREM-26, reference 314902 from BioLegend), the purified anti-human
CD354 (TREM-1) antibody (clone TREM-37, reference 316102 from
BioLegend), the monoclonal mouse anti-human sTREM1 (clone 15G7,
reference 298099 from USBio), the mouse anti-human TREM1 (clone
2E2, reference 134704 from USBio). Other non-limitative examples of
antibodies allowing the detection of sTREM-1 include sTREM-1 and/or
TREM-1 antibodies described in the following patents or patent
applications: US2013/150559, US 2013/211050, US 2013/309239,
WO2013/120553 and U.S. Pat. No. 8,106,165.
[0164] Some of the aforementioned assays for measuring the level of
sTREM-1 in a biological sample (such as, for example, western blot,
ELISA, or sandwich ELISA) generally involve the binding of the
partner (i.e., an antibody or an aptamer) to a solid support. Solid
supports which can be used in the practice of the method of the
invention include, without being limited to, supports such as
nitrocellulose (e.g., nitrocellulose membranes or nitrocellulose
microtiter plates); polyvinylchloride (e.g., polyvinylchloride
sheets, polyvinylchloride membranes or polyvinylchloride microtiter
plates); polystyrene latex (e.g., polystyrene latex beads or
polystyrene latex microtiter plates); polyvinylidene difluoride or
PVDF (e.g., PVDF membranes); diazotized paper; nylon membranes;
activated beads, and magnetically responsive beads.
[0165] The level of sTREM-1 may be measured by using standard
immunodiagnostic techniques, including immunoassays such as
competition, direct reaction, or sandwich type assays. Such assays
include, but are not limited to, agglutination tests;
enzyme-labelled and mediated immunoassays, such as ELISAs;
biotin/avidin type assays; radioimmunoassays;
immuno-electrophoresis; immuno-precipitation Immunoassays thus
include, without being limited to, enzyme-labelled and mediated
immunoassays, such as ELISAs or enzyme-linked fluorescent assays
(ELFA); biotin/avidin type assays; radioimmunoassays;
immuno-electrophoresis; immuno-precipitation;
electrochemiluminescence immunoassay (ECLIA).
[0166] An exemplary biochemical test for identifying specific
proteins employs a standardized test format, such as ELISA test,
although the information provided herein may apply to the
development of other biochemical tests and is not limited to the
development of an ELISA test (see for example Molecular Immunology:
A Textbook, edited by Atassi et al. Marcel Dekker Inc., New York
and Basel 1984, for a description of ELISA tests). It is understood
that commercial assay enzyme-linked immunosorbent assay (ELISA)
kits for various plasma constituents are available.
[0167] Therefore, the ELISA method may be used for measuring the
level of sTREM-1 in a biological sample, wherein the wells of a
microtiter plate are coated with at least one antibody which
recognizes sTREM-1. A biological sample containing or suspected of
containing sTREM-1 is then added to the coated wells. After a
period of incubation sufficient to allow the formation of
antibody-sTREM-1 complexes, the plate(s) can be washed to remove
unbound moieties and a detectably labelled secondary binding
molecule added. The secondary binding molecule is allowed to react
with any captured antibody-sTREM-1 complexes, the plate washed and
the presence of the secondary binding molecule detected using
methods well-known in the art.
[0168] In one embodiment, measuring the level of sTREM-1 in a
biological sample as described hereinabove comprises the use of an
enzyme-linked immunosorbent assay (ELISA), an
electrochemiluminescence immunoassay (ECLIA) (such as Elecsys.RTM.
Roche Diagnostics) or an enzyme-linked fluorescent assay
(ELFA).
[0169] Examples of ELISA assays include, without being limited to,
the TREM-1 Quantikine ELISA kit (reference DTRM10C from R&D
Systems); the human TREM-1 DuoSet (references DY1278B and DY1278BE
from R&D Systems), the sTREM-1 ELISA (reference sTREM-1 ELISA
from iQProducts).
[0170] Examples of electrochemiluminescence immunoassays (ECLIAs)
include Elecsys.RTM. (Roche Diagnostics).
[0171] Examples of enzyme-linked fluorescent assays (ELFAs) include
VIDAS.RTM. (Biomerieux).
[0172] Typically, the level of sTREM-1 in a biological sample may
be measured by an immunometric assay on the basis of a
double-antibody "sandwich" technique, with a monoclonal antibody
specific for sTREM-1. Thus, in one embodiment, measuring the level
of sTREM-1 in a biological sample as described hereinabove
comprises the use sandwich ELISA.
[0173] According to one embodiment, measuring the level of sTREM-1
(with or without immunoassay-based methods) may also include
separation of the compounds present in the biological sample (i.e.,
separation of sTREM-1): centrifugation based on the compound's
molecular weight; electrophoresis based on mass and charge; HPLC
based on hydrophobicity; size exclusion chromatography based on
size; and solid-phase affinity based on the compound's affinity for
the particular solid-phase that is used.
[0174] Once separated, said compounds (i.e., sTREM-1) may be
identified based on the known "separation profile", e.g., retention
time, for that compound and measured using standard techniques.
Alternatively, the separated compounds (i.e., sTREM-1) may be
detected and measured by, for example, a mass spectrometer.
[0175] According to one embodiment, the level of sTREM-1 refers to
a nucleic acid level, a nucleic acid quantity, a nucleic acid
amount or a nucleic acid concentration. In one embodiment, the
nucleic acid is an RNA, preferably an mRNA, or a cDNA.
[0176] Methods for measuring the expression level, in particular a
sTREM-1 nucleic acid level, in a biological sample as described
hereinabove are well-known to the skilled artisan and include,
without being limited to, PCR, qPCR, RT-PCR, RT-qPCR, Northern
Blot, hybridization techniques such as, for example, use of
microarrays, and combination thereof including but not limited to,
hybridization of amplicons obtained by RT-PCR, sequencing such as,
for example, next-generation DNA sequencing (NGS) or RNA-seq (also
known as "Whole Transcriptome Shotgun Sequencing").
[0177] In one embodiment, the sTREM-1 nucleic acid level is
measured using the forward and reverse primers having a nucleotide
sequence has set forth in SEQ ID NO: 13 (GTGGTGACCAAGGGGTTC) and
SEQ ID NO: 14 (AGATGGATGTGGCTG GAAGT), respectively.
[0178] In one embodiment, the sTREM-1 nucleic acid level is
measured using the forward and reverse primers having a nucleotide
sequence has set forth in SEQ ID NO: 15 (GTGACCAAGGGTTTTTCAGG) and
SEQ ID NO: 16 (ACACCGGAACCCTGAT GATA), respectively.
[0179] In one embodiment, the sTREM-1 nucleic acid level is
measured using the forward and reverse primers having a nucleotide
sequence has set forth in SEQ ID NO: 17 (AAAGGCAAGAACGCCTGAC) and
SEQ ID NO: 18 (GGGACTTTACCAAGAGG GAC), respectively
[0180] In one embodiment, the level of sTREM-1 measured in a
biological sample as described hereinabove is a baseline level. In
other words, the level of sTREM-1 is measured in a biological
sample as described hereinabove obtained from the human subject
suffering from an inflammatory disorder before the beginning of an
administration of a therapy, preferably of a TREM-1 inhibitor.
[0181] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove obtained from the human
subject between the first 2 hours and the first 48 hours,
preferably between the first 2 hours and the first 12 hours,
between the first 12 hours and the first 24 hours or between the
first 24 hours and the first 48 hours, following the onset of an
inflammatory disorder, preferably SIRS, sepsis or septic shock;
following the diagnosis of the human subject with an inflammatory
disorder, preferably SIRS, sepsis or septic shock; following the
hospitalization, in particular the admission in ICU or emergency
unit, of the human subject for an inflammatory disorder, preferably
SIRS, sepsis or septic shock; or following the start of vasopressor
therapy.
[0182] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove obtained from the human
subject within the first 2, 3, 6, 9, 12, 15, 18, 21, 24, 30, 36, 42
or 48 hours following the onset of an inflammatory disorder,
preferably SIRS, sepsis or septic shock; following the diagnosis of
the human subject with an inflammatory disorder, preferably SIRS,
sepsis or septic shock; following the hospitalization, in
particular the admission in ICU or emergency unit, of the human
subject for an inflammatory disorder, preferably SIRS, sepsis or
septic shock; or following the start of vasopressor therapy.
[0183] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove between the first 2
hours and the first 48 hours, preferably between the first 2 hours
and the first 12 hours, between the first 12 hours and the first 24
hours or between the first 24 hours and the first 48 hours,
following the onset of an inflammatory disorder, preferably SIRS,
sepsis or septic shock; following the diagnosis of the human
subject with an inflammatory disorder, preferably SIRS, sepsis or
septic shock; following the hospitalization, in particular the
admission in ICU or emergency unit, of the human subject for an
inflammatory disorder, preferably SIRS, sepsis or septic shock; or
following the start of vasopressor therapy.
[0184] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove within the first 2, 3,
6, 9, 12, 15, 18, 21, 24, 30, 36, 42 or 48 hours following the
onset of an inflammatory disorder, preferably SIRS, sepsis or
septic shock; following the diagnosis of the human subject with an
inflammatory disorder, preferably SIRS, sepsis or septic shock;
following the hospitalization, in particular the admission in ICU
or emergency unit, of the human subject for an inflammatory
disorder, preferably SIRS, sepsis or septic shock; or following the
start of vasopressor therapy.
[0185] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove obtained from the human
subject between the first 2 hours and the first 48 hours,
preferably between the first 2 hours and the first 12 hours,
between the first 12 hours and the first 24 hours or between the
first 24 hours and the first 48 hours, following a reinfection of
the human subject suffering from an inflammatory disorder,
preferably SIRS, sepsis or septic shock.
[0186] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove obtained from the human
subject within the first 2, 3, 6, 9, 12, 15, 18, 21, 24, 30, 36, 42
or 48 hours following a reinfection of the human subject suffering
from an inflammatory disorder, preferably SIRS, sepsis or septic
shock.
[0187] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove between the first 2
hours and the first 48 hours, preferably between the first 2 hours
and the first 12 hours, between the first 12 hours and the first 24
hours or between the first 24 hours and the first 48 hours,
following a reinfection of the human subject suffering from an
inflammatory disorder, preferably SIRS, sepsis or septic shock.
[0188] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove within the first 2, 3,
6, 9, 12, 15, 18, 21, 24, 30, 36, 42 or 48 hours following a
reinfection of the human subject suffering from an inflammatory
disorder, preferably SIRS, sepsis or septic shock.
[0189] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove obtained from the human
subject between the first 2 hours and the first 48 hours,
preferably between the first 2 hours and the first 12 hours,
between the first 12 hours and the first 24 hours or between the
first 24 hours and the first 48 hours, following a
rehospitalization, in particular a readmission to ICU or emergency
unit, of the human subject suffering from an inflammatory disorder,
preferably SIRS, sepsis or septic shock.
[0190] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove obtained from the human
subject within the first 2, 3, 6, 9, 12, 15, 18, 21, 24, 30, 36, 42
or 48 hours following a rehospitalization, in particular a
readmission to ICU or emergency unit, of the human subject
suffering from an inflammatory disorder, preferably SIRS, sepsis or
septic shock.
[0191] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove between the first 2
hours and the first 48 hours, preferably between the first 2 hours
and the first 12 hours, between the first 12 hours and the first 24
hours or between the first 24 hours and the first 48 hours,
following a rehospitalization, in particular a readmission to ICU
or emergency unit, of the human subject suffering from an
inflammatory disorder, preferably SIRS, sepsis or septic shock.
[0192] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove within the first 2, 3,
6, 9, 12, 15, 18, 21, 24, 30, 36, 42 or 48 hours following a
rehospitalization, in particular a readmission to ICU or emergency
unit, of the human subject suffering from an inflammatory disorder,
preferably SIRS, sepsis or septic shock.
[0193] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove obtained from the human
subject between the first 2 hours and the first 48 hours,
preferably between the first 2 hours and the first 12 hours,
between the first 12 hours and the first 24 hours or between the
first 24 hours and the first 48 hours following the onset of septic
shock; following the diagnosis of the human subject for septic
shock; following the hospitalization, in particular the admission
to ICU or emergency unit, of the human subject for septic shock; or
following the start of vasopressor therapy.
[0194] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove obtained from the human
subject within the first 2, 3, 6, 9, 12, 15, 18, 21, 24, 30, 36, 42
or 48 hours following the onset of septic shock; following the
diagnosis of the human subject for septic shock; following the
hospitalization, in particular the admission to ICU or emergency
unit, of the human subject for septic shock; or following the start
of vasopressor therapy.
[0195] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove between the first 2
hours and the first 12 hours, the first 12 hours and the first 24
hours or between the first 24 hours and the first 48 hours
following the onset of septic shock; following the diagnosis of the
human subject for septic shock; following the hospitalization, in
particular the admission to ICU or emergency unit, of the human
subject for septic shock; or following the start of vasopressor
therapy.
[0196] In one embodiment, the level of sTREM-1 is measured in a
biological sample as described hereinabove within the first 2, 3,
6, 9, 12, 15, 18, 21, 24, 30, 36, 42 or 48 hours following the
onset of septic shock; following the diagnosis of the human subject
for septic shock; following the hospitalization, in particular the
admission to ICU or emergency unit, of the human subject for septic
shock; or following the start of vasopressor therapy.
[0197] In one embodiment, the predetermined sTREM-1 value is a
personalized reference value, i.e., the predetermined sTREM-1 value
is obtained using a biological sample obtained from the human
subject.
[0198] In one embodiment, the predetermined value of sTREM-1 is an
index value or is derived from one or more risk prediction
algorithms or computed indices for an inflammatory disorder,
preferably SIRS, sepsis or septic shock.
[0199] In one embodiment, the predetermined value of sTREM-1 is
obtained from a reference population.
[0200] According to the present invention, the predetermined
sTREM-1 value can be derived from population studies, including,
for example, subjects having a similar age range, subjects in the
same or similar ethnic group, subjects having a chronic medical
condition linked with an increased risk of an inflammatory
disorder, preferably SIRS, sepsis or septic shock (such as atrial
fibrillation, cancer, chronic kidney disease, chronic lung disease,
cirrhosis, coronary artery disease, deep vein thrombosis, diabetes,
dyslipidemia, endocarditis, hypertension, influenza, malaria or any
other protozoan parasitic disease, myocardial infarction,
neurological disease, non-alcoholic fatty liver disease (NAFLD),
non-alcoholic steatohepatitis (NASH), peripheral artery disease,
pulmonary fibrosis, severe obesity and stroke) or subjects with a
history of an inflammatory disorder such as SIRS, sepsis or septic
shock.
[0201] According to one embodiment, the predetermined value of
sTREM-1 is derived from the measure of the sTREM-1 level in a
biological sample from one or more human subjects who are
substantially healthy. As used herein, a "substantially healthy
subject" is a human subject who has not been previously diagnosed
or identified as having or suffering from an inflammatory disorder,
preferably SIRS, sepsis or septic shock. Thus, according to one
embodiment, the predetermined sTREM-1 value is obtained from a
reference population of human subjects who are substantially
healthy.
[0202] In one embodiment, a "substantially healthy subject" is a
human subject who does not suffer from an infection.
[0203] According to one embodiment, the predetermined value of
sTREM-1 obtained from a reference population of human subjects who
are substantially healthy is a sTREM-1 level, preferably a blood,
plasma or serum level, ranging from about 50 pg/mL to about 250
pg/mL.
[0204] In one embodiment, the predetermined value of sTREM-1
obtained from a reference population of human subjects who are
substantially healthy is a sTREM-1 level, preferably a blood,
plasma or serum level, of about 50, 75, 100, 125, 150, 175, 200,
225 or 250 pg/mL.
[0205] In one embodiment, the predetermined value of sTREM-1
obtained from a reference population of human subjects who are
substantially healthy is a sTREM-1 level, preferably a blood,
plasma or serum level, ranging from about 50 pg/mL to about 150
pg/mL, in particular as determined using an enzyme-linked
immunosorbent assay (ELISA).
[0206] In one embodiment, the predetermined value of sTREM-1
obtained from a reference population of human subjects who are
substantially healthy is a sTREM-1 level, preferably a blood,
plasma or serum level, of about 50, 75, 100, 125 or 150 pg/mL, in
particular as determined using an enzyme-linked immunosorbent assay
(ELISA).
[0207] In one embodiment, the predetermined value of sTREM-1
obtained from a reference population of human subjects who are
substantially healthy is a sTREM-1 level, preferably a blood,
plasma or serum level, ranging from about 150 pg/mL to about 250
pg/mL, in particular as determined using an
electrochemiluminescence immunoassay (ECLIA).
[0208] In one embodiment, the predetermined value of sTREM-1
obtained from a reference population of human subjects who are
substantially healthy is a sTREM-1 level, preferably a blood,
plasma or serum level, of about 150, 175, 200, 225 or 250 pg/mL, in
particular as determined using an electrochemiluminescence
immunoassay (ECLIA).
[0209] According to another embodiment, the predetermined value of
sTREM-1 is derived from the measure of the sTREM-1 level in a
biological sample from one or more human subjects diagnosed or
identified as suffering, or having suffered, from an inflammatory
disorder, preferably SIRS, sepsis or septic shock. Thus, according
to one embodiment, the predetermined sTREM-1 value is obtained from
a reference population of human subjects diagnosed or identified as
suffering, or having suffered, from an inflammatory disorder,
preferably SIRS, sepsis or septic shock.
[0210] In one embodiment, the predetermined value of sTREM-1 is
derived from the measure of the sTREM-1 level in a biological
sample from one or more human subjects diagnosed or identified as
suffering from sepsis. Thus, in one embodiment, the predetermined
sTREM-1 value is obtained from a reference population of human
subjects diagnosed or identified as suffering from sepsis.
[0211] In one embodiment, the in vitro method of the invention is
for identifying a human subject suffering from sepsis susceptible
to respond to an administration of a therapy, in particular of a
TREM-1 inhibitor, and the predetermined value of sTREM-1 is derived
from the measure of the sTREM-1 level in a biological sample from
one or more human subjects diagnosed or identified as suffering
from sepsis. In other words, in one embodiment, the in vitro method
of the invention is for identifying a human subject suffering from
sepsis susceptible to respond to an administration of a therapy, in
particular of a TREM-1 inhibitor, and the predetermined sTREM-1
value is obtained from a reference population of human subjects
diagnosed or identified as suffering from sepsis.
[0212] In one embodiment, the predetermined value of sTREM-1 is
derived from the measure of the sTREM-1 level in a biological
sample from one or more human subjects diagnosed or identified as
suffering from septic shock. Thus, in one embodiment, the
predetermined sTREM-1 value is obtained from a reference population
of human subjects diagnosed or identified as suffering from septic
shock.
[0213] In one embodiment, the in vitro method of the invention is
for identifying a human subject suffering from septic shock
susceptible to respond to an administration of a therapy, in
particular of a TREM-1 inhibitor, and the predetermined value of
sTREM-1 is derived from the measure of the sTREM-1 level in a
biological sample from one or more human subjects diagnosed or
identified as suffering from septic shock. In other words, in one
embodiment, the in vitro method of the invention is for identifying
a human subject suffering from septic shock susceptible to respond
to an administration of a therapy, in particular of a TREM-1
inhibitor, and the predetermined sTREM-1 value is obtained from a
reference population of human subjects diagnosed or identified as
suffering from septic shock.
[0214] In one embodiment, the predetermined value of sTREM-1 is
derived from the measure of the sTREM-1 level in a biological
sample from one or more human subjects who have previously been
diagnosed or identified as suffering from an inflammatory disorder,
preferably SIRS, sepsis or septic shock.
[0215] In one embodiment, the predetermined value of sTREM-1 is
derived from the measure of the sTREM-1 level in a biological
sample from one or more human subjects suffering from an infection
who are at high risk of developing an inflammatory disorder,
preferably SIRS, sepsis or septic shock.
[0216] According to the present invention, the predetermined
sTREM-1 value can be derived from statistical analyses and/or risk
prediction data of a reference population as described hereinabove
obtained from mathematical algorithms and computed indices of an
inflammatory disorder, preferably SIRS, sepsis or septic shock.
[0217] According to one embodiment, the predetermined value of
sTREM-1 is obtained from a reference population as described
hereinabove using a method of statistical and/or structural
classification.
[0218] According to one embodiment, the predetermined sTREM-1
value, preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level.
[0219] In one embodiment, a human subject suffering from an
inflammatory disorder susceptible to respond to a therapy is a
human subject with a level of sTREM-1 measured as described
hereinabove higher than at least 1.5 times, 2 times, 2.5 times, 3
times, 3.5 times, 4 times, 4.5 times, 5 times, 5.5 times, 6 times,
6.5 times, 7 times, 7.5 times, 8 times, 8.5 times, 9 times, 9.5
times or 10 times the predetermined sTREM-1, preferably the
predetermined sTREM-1 obtained from a reference population of human
subjects who are substantially healthy, as described
hereinabove.
[0220] In one embodiment, a human subject suffering from an
inflammatory disorder susceptible to respond to a therapy is a
human subject with a level of sTREM-1 measured as described
hereinabove higher than at least 1.6 times, 1.8 times, 2 times, 2.2
times, 2.4 times, 2.6 times, 2.8 times, 3 times, 3.2 times, 3.4
times, 3.6 times, 3.8 times, 4 times, 4.2 times, 4.4 times, 4.6
times, 4.8 times, 5 times, 5.2 times, 5.4 times, 5.6 times, 5.8
times or 6 times the predetermined sTREM-1, preferably the
predetermined sTREM-1 obtained from a reference population of human
subjects who are substantially healthy, as described
hereinabove.
[0221] In one embodiment, a human subject suffering from an
inflammatory disorder susceptible to respond to a therapy is a
human subject with a level of sTREM-1 measured as described
hereinabove higher than at least 2 times, 2.1 times, 2.2 times, 2.3
times, 2.4 times, 2.5 times, 2.6 times, 2.7 times, 2.8 times, 2.9
times, 3 times, 3.1 times, 3.2 times, 3.3 times, 3.4 times, 3.5
times, 3.6 times, 3.7 times, 3.8 times, 3.9 times or 4 times the
predetermined sTREM-1, preferably the predetermined sTREM-1
obtained from a reference population of human subjects who are
substantially healthy, as described hereinabove.
[0222] In another embodiment, a human subject suffering from an
inflammatory disorder susceptible to respond to a therapy is a
human subject with a level of sTREM-1 measured as described
hereinabove higher than at least 1.1 times, 1.2 times, 1.3 times,
1.4 times, 1.5 times, 1.6 times, 1.7 times, 1.8 times, 1.9 times, 2
times, 2.1 times, 2.2 times, 2.3 times, 2.4 times or 2.5 times the
predetermined sTREM-1, preferably the predetermined sTREM-1
obtained from a reference population of human subjects diagnosed or
identified as suffering, or having suffered, from an inflammatory
disorder, preferably SIRS, sepsis or septic shock.
[0223] In one embodiment, the predetermined sTREM-1 value obtained
from a reference population as described hereinabove, is the
average sTREM-1 level, preferably the average blood, plasma or
serum level, of said reference population. In another embodiment,
the predetermined sTREM-1 value obtained from a reference
population as described hereinabove, is the median sTREM-1 level,
preferably the average blood, plasma or serum level, of said
reference population.
[0224] According to one embodiment, the predetermined sTREM-1 value
is obtained from a reference population as described hereinabove,
wherein the sTREM-1 levels measured in a biological sample from
each of the human subjects of the reference population (i.e., the
sTREM-1 levels measured in the reference population) are divided
into equal-sized groups by cut-off values referred to as
"quantiles", each group corresponding to a determined percentage of
the sTREM-1 levels measured in the reference population. Examples
of quantiles include, without being limited to, the median
(defining 2 groups each comprising 50% of the sTREM-1 levels
measured in the reference population), the terciles or tertiles
(defining 3 groups each comprising a third of the sTREM-1 levels
measured in the reference population), the quartiles (defining 4
groups each comprising 25% of the sTREM-1 levels measured in the
reference population), the quintiles (defining 5 groups each
comprising 20% of the sTREM-1 levels measured in the reference
population) and the deciles (defining 10 groups each comprising 10%
of the sTREM-1 levels measured in the reference population).
[0225] According to the present invention, "quantiles" refer to the
cut-off sTREM-1 values below or above which lies a determined
percentage of the sTREM-1 levels measured in the reference
population. Therefore, the human subjects with a measured sTREM-1
level below the first quantile are the human subjects with the
lowest sTREM-1 levels, while the human subjects with a measured
sTREM-1 level above the last quantile are the human subjects with
the highest sTREM-1 levels. For example, the 1.sup.st decile is the
sTREM-1 value below which 10% of the sTREM-1 levels measured in the
reference population lie and above which 90% of the sTREM-1 levels
measured in the reference population lie.
[0226] Additionally, the term "quantiles" may also sometimes refer
to the group so defined by said cut-off value. Thus, applied to the
present invention, the term "quantiles" may also refer to the
groups of sTREM-1 levels measured in the reference population
defined by the cut-off sTREM-1 value. For example, the 1.sup.st
decile may refer to the group of sTREM-1 levels measured in the
reference population corresponding to the lowest 10% of sTREM-1
levels measured in the reference population. Accordingly, the
10.sup.th decile refers to the group of sTREM-1 levels measured in
the reference population corresponding to the highest 10% of
sTREM-1 levels measured in the reference population. It follows
that a sTREM-1 value that is in the 1.sup.st decile is a sTREM-1
value comprised in the lowest 10% of sTREM-1 levels measured in the
reference population and that a sTREM-1 value that is in the
10.sup.th decile is a sTREM-1 value comprised in the highest 10% of
sTREM-1 levels measured in the reference population.
[0227] In one embodiment, the predetermined sTREM-1 value is
obtained from a reference population as described hereinabove,
wherein the sTREM-1 levels measured in the reference population are
divided into two equal-sized groups each corresponding to 50% of
the sTREM-1 levels measured in the reference population.
[0228] According to this embodiment of the present invention the
sTREM-1 median corresponds to the sTREM-1 value below which 50% of
the sTREM-1 levels measured in the reference population lie and
above which 50% of the sTREM-1 levels measured in the reference
population lie.
[0229] Thus, in one embodiment, the predetermined sTREM-1 value is
the sTREM-1 median of a reference population as described
hereinabove.
[0230] Accordingly, in one embodiment, the present invention
relates to an in vitro method for identifying a human subject
suffering from an inflammatory disorder, preferably SIRS, sepsis or
septic shock, susceptible to respond to an administration of a
therapy, in particular of a TREM-1 inhibitor, said method
comprising: [0231] a) measuring the level of soluble Triggering
Receptors Expressed on Myeloid cells-1 (sTREM-1) in a biological
sample from the human subject; [0232] b) comparing the level of
sTREM-1 measured at step a) to a predetermined sTREM-1 value
obtained from a reference population sTREM-1, said predetermined
sTREM-1 value being the median of said reference population; [0233]
c) identifying a human subject suffering from an inflammatory
disorder, preferably SIRS, sepsis or septic shock with a level of
sTREM-1 measured at step a) higher than the median of step b) as
susceptible to respond to an administration of a therapy, in
particular of a TREM-1 inhibitor.
[0234] In one embodiment, the predetermined sTREM-1 value is
obtained from a reference population as described hereinabove,
wherein the sTREM-1 levels measured in the reference population are
divided into three equal-sized groups each corresponding to a third
of the sTREM-1 levels measured in the reference population. As
mentioned above, the cut-off values ("quantiles") so dividing the
sTREM-1 levels measured in the reference population are called
"terciles" (or "tertiles"). Thus, in one embodiment, the
predetermined sTREM-1 value is a sTREM-1 tercile (or tertile) of a
reference population as described hereinabove.
[0235] According to this embodiment of the present invention:
[0236] the sTREM-1 first tercile (or tertile) corresponds to the
sTREM-1 value below which a third of the sTREM-1 levels measured in
the reference population lie and above which two thirds of the
sTREM-1 levels measured in the reference population lie; and [0237]
the sTREM-1 second tercile (or tertile) corresponds to the sTREM-1
value below which two thirds of the sTREM-1 levels measured in the
reference population lie and above which one third of the sTREM-1
levels measured in the reference population lie.
[0238] In one embodiment, the predetermined sTREM-1 value is the
second sTREM-1 tercile (i.e., the last sTREM-1 tercile) of a
reference population as described hereinabove.
[0239] In one embodiment, the predetermined sTREM-1 value is
obtained from a reference population as described hereinabove,
wherein the sTREM-1 levels measured in the reference population are
divided into four equal-sized groups each corresponding 25% of the
sTREM-1 levels measured in the reference population. As mentioned
above, the cut-off values ("quantiles") so dividing the sTREM-1
levels measured in the reference population are called "quartiles".
Thus, in one embodiment, the predetermined sTREM-1 value is a
sTREM-1 quartile of a reference population as described
hereinabove.
[0240] According to this embodiment of the present invention:
[0241] the sTREM-1 first quartile (or Q1) corresponds to the
sTREM-1 value below which 25% of the sTREM-1 levels measured in the
reference population lie and above which 75% of the sTREM-1 levels
measured in the reference population lie; [0242] the sTREM-1 second
quartile corresponds to the sTREM-1 value below which 50% of the
sTREM-1 levels measured in the reference population lie and above
which 50% of the sTREM-1 levels measured in the reference
population lie (the second quartile is thus equivalent to the
median); [0243] the sTREM-1 third quartile (or Q3) corresponds to
the sTREM-1 value below which 75% of the sTREM-1 levels measured in
the reference population lie and above which 25% of the sTREM-1
levels measured in the reference population lie.
[0244] In one embodiment, the predetermined sTREM-1 value is the
first sTREM-1 quartile also referred to as sTREM-1 Q1 of a
reference population as described hereinabove.
[0245] In one embodiment, the predetermined sTREM-1 value is the
third sTREM-1 quartile also referred to as sTREM-1 Q3 (i.e., the
last sTREM-1 quartile) of a reference population as described
hereinabove.
[0246] Accordingly, in one embodiment, the present invention
relates to an in vitro method for identifying a human subject
suffering from an inflammatory disorder, preferably SIRS, sepsis or
septic shock, susceptible to respond to an administration of a
therapy, in particular of a TREM-1 inhibitor, said method
comprising: [0247] a) measuring the level of soluble Triggering
Receptors Expressed on Myeloid cells-1 (sTREM-1) in a biological
sample from the human subject; [0248] b) comparing the level of
sTREM-1 measured at step a) to a predetermined sTREM-1 value
obtained from a reference population sTREM-1, said predetermined
sTREM-1 value being the third quartile of said reference
population; [0249] c) identifying a human subject suffering from an
inflammatory disorder, preferably SIRS, sepsis or septic shock with
a level of sTREM-1 measured at step a) higher than the third
quartile of step b) as susceptible to respond to an administration
of a therapy, in particular of a TREM-1 inhibitor.
[0250] In one embodiment, the predetermined sTREM-1 value is
obtained from a reference population as described hereinabove,
wherein the sTREM-1 levels measured in the reference population are
divided into five equal-sized groups each corresponding to 20% of
the sTREM-1 levels measured in the reference population. As
mentioned above, the cut-off values ("quantiles") so dividing the
sTREM-1 levels measured in the reference population are called
"quintiles". Thus, in one embodiment, the predetermined sTREM-1
value is a sTREM-1 quintile of a reference population as described
hereinabove.
[0251] According to this embodiment of the present invention:
[0252] the sTREM-1 first quintile corresponds to the sTREM-1 value
below which 20% of the sTREM-1 levels measured in the reference
population lie and above which 80% of the sTREM-1 levels measured
in the reference population lie; [0253] the sTREM-1 second quintile
corresponds to the sTREM-1 value below which 40% of the sTREM-1
levels measured in the reference population lie and above which 60%
of the sTREM-1 levels measured in the reference population lie;
[0254] the sTREM-1 third quintile corresponds to the sTREM-1 value
below which 60% of the sTREM-1 levels measured in the reference
population lie and above which 40% of the sTREM-1 levels measured
in the reference population lie; and [0255] the sTREM-1 fourth
quintile corresponds to the sTREM-1 value below which 80% of the
sTREM-1 levels measured in the reference population lie and above
which 20% of the sTREM-1 levels measured in the reference
population lie.
[0256] In one embodiment, the predetermined sTREM-1 value is the
second sTREM-1 quintile of a reference population as described
hereinabove.
[0257] In one embodiment, the predetermined sTREM-1 value is the
third sTREM-1 quintile (i.e., the penultimate sTREM-1 quintile) of
a reference population as described hereinabove.
[0258] In one embodiment, the predetermined sTREM-1 value is the
fourth sTREM-1 quintile (i.e., the last sTREM-1 quintile) of a
reference population as described hereinabove.
[0259] In one embodiment, the predetermined sTREM-1 value is
obtained from a reference population as described hereinabove,
wherein the sTREM-1 levels measured in the reference population are
divided into ten equal-sized groups each corresponding to 10% of
the sTREM-1 levels measured in the reference population. As
mentioned above, the cut-off values ("quantiles") so dividing the
sTREM-1 levels measured in the reference population are called
"deciles". Thus, in one embodiment, the predetermined sTREM-1 value
is a sTREM-1 decile of a reference population as described
hereinabove.
[0260] According to this embodiment of the present invention:
[0261] the sTREM-1 first decile corresponds to the sTREM-1 value
below which 10% of the sTREM-1 levels measured in the reference
population lie and above which 90% of the sTREM-1 levels measured
in the reference population lie; [0262] the sTREM-1 second decile
corresponds to the sTREM-1 value below which 20% of the sTREM-1
levels measured in the reference population lie and above which 80%
of the sTREM-1 levels measured in the reference population lie;
[0263] the sTREM-1 third decile corresponds to the sTREM-1 value
below which 30% of the sTREM-1 levels measured in the reference
population lie and above which 70% of the sTREM-1 levels measured
in the reference population lie; [0264] the sTREM-1 fourth decile
corresponds to the sTREM-1 value below which 40% of the sTREM-1
levels measured in the reference population lie and above which 60%
of the sTREM-1 levels measured in the reference population lie;
[0265] the sTREM-1 fifth decile corresponds to the sTREM-1 value
below which 50% of the sTREM-1 levels measured in the reference
population lie and above which 50% of the sTREM-1 levels measured
in the reference population lie; [0266] the sTREM-1 sixth decile
corresponds to the sTREM-1 value below which 60% of the sTREM-1
levels measured in the reference population lie and above which 40%
of the sTREM-1 levels measured in the reference population lie;
[0267] the sTREM-1 seventh decile corresponds to the sTREM-1 value
below which 70% of the sTREM-1 levels measured in the reference
population lie and above which 30% of the sTREM-1 levels measured
in the reference population lie; [0268] the sTREM-1 eight decile
corresponds to the sTREM-1 value below which 80% of the sTREM-1
levels measured in the reference population lie and above which 20%
of the sTREM-1 levels measured in the reference population lie;
[0269] the sTREM-1 ninth decile corresponds to the sTREM-1 value
below which 90% of the sTREM-1 levels measured in the reference
population lie and above which 10% of the sTREM-1 levels measured
in the reference population lie;
[0270] In one embodiment, the predetermined sTREM-1 value is the
sixth sTREM-1 decile of a reference population as described
hereinabove. In one embodiment, the predetermined sTREM-1 value is
the seventh sTREM-1 decile of a reference population as described
hereinabove. In one embodiment, the predetermined sTREM-1 value is
the eight sTREM-1 decile (i.e., the penultimate sTREM-1 decile) of
a reference population as described hereinabove.
[0271] In one embodiment, the predetermined sTREM-1 value is the
ninth sTREM-1 decile (i.e., the last sTREM-1 decile) of a reference
population as described hereinabove.
[0272] In one embodiment, the predetermined sTREM-1 value is
obtained from a reference population as described hereinabove,
wherein the sTREM-1 levels measured in the reference population are
divided into groups corresponding to a given percentage of the
sTREM-1 levels measured in the reference population. As mentioned
above, the cut-off values ("quantiles") so dividing the sTREM-1
levels measured in the reference population are called
"percentiles". Thus, in one embodiment, the predetermined sTREM-1
value is a sTREM-1 percentile of a reference population as
described hereinabove.
[0273] According to one embodiment, the predetermined sTREM-1 value
is obtained from a reference population as described hereinabove,
wherein said value is associated with a predicted mortality rate in
the reference population of 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,
60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%.
[0274] For example, a predetermined sTREM-1 value associated with a
predicted mortality rate in the reference population of 50% means
that for subjects of the reference population having a sTREM-1
level higher than said value, the mortality rate is of 50%.
[0275] According to one embodiment, the predetermined sTREM-1 value
is obtained from a reference population as described hereinabove
and comprising patients affected with SIRS, sepsis or septic shock,
wherein, in said reference population, more than 20%, 25%, 30%,
35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%
of the subjects dying in the 28, 90 or 365 days following the first
infection or the first hospitalization present a value higher than
said the predetermined sTREM-1 value.
[0276] According to one embodiment, the predetermined sTREM-1
value, preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 20
pg/mL to about 6000 pg/mL.
[0277] In one embodiment the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 20, 50, 100,
150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750,
800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350,
1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900,
1950, 2000, 2050, 2100, 2150, 2200, 2250, 2300, 2350, 2400, 2450,
2500, 2550, 2600, 2650, 2700, 2750, 2800, 2850, 2900, 2950, 3000,
3050, 3100, 3150, 3200, 3250, 3300, 3350, 3400, 3450, 3500, 3550,
3600, 3650, 3700, 3750, 3800, 3850, 3900, 3950, 4000, 4050, 4100,
4150, 4200, 4250, 4300, 4350, 4400, 4450, 4500, 4550, 4600, 4650,
4700, 4750, 4800, 4850, 4900, 4950, 5000, 5050, 5100, 5150, 5200,
5250, 5300, 5350, 5400, 5450, 5500, 5550, 5600, 5650, 5700, 5750,
5800, 5850, 5900, 5950 or 6000 pg/mL.
[0278] According to one embodiment, the predetermined sTREM-1
value, preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 30
pg/mL to about 2000 pg/mL.
[0279] In one embodiment the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 30, 50, 100,
150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750,
800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350,
1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900,
1950 or 2000 pg/mL.
[0280] According to one embodiment, the predetermined sTREM-1
value, preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 50
pg/mL to about 1000 pg/mL.
[0281] In one embodiment the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 50, 75, 100,
125, 150, 175, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290,
300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420,
430, 440, 450, 460, 470, 480, 490, 500, 520, 540, 560, 580, 600,
620, 640, 660, 680, 700, 720, 740, 760, 780, 800, 820, 840, 860,
880, 900, 920, 940, 960, 980 or 1000 pg/mL.
[0282] According to another embodiment, the predetermined sTREM-1
value, preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 250
pg/mL to about 400 pg/mL.
[0283] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 250, 255, 260,
265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325,
330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390,
395 or 400 pg/mL.
[0284] According to another embodiment, the predetermined sTREM-1
value, preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 300
pg/mL to about 800 pg/mL.
[0285] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 300, 305, 310,
315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375,
380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440,
445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 505,
510, 515, 520, 525, 530, 535, 540, 545, 550, 555, 560, 565, 570,
575, 580, 585, 590, 595, 600, 605, 610, 615, 620, 625, 630, 635,
640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690, 695, 700,
705, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760, 765,
770, 775, 780, 785, 790, 795 or 800 pg/mL.
[0286] According to another embodiment, the predetermined sTREM-1
value, preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 350
pg/mL to about 600 pg/mL.
[0287] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 350, 355, 360,
365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425,
430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490,
495, 500, 505, 510, 515, 520, 525, 530, 535, 540, 545, 550, 555,
560, 565, 570, 575, 580, 585, 590, 595 or 600 pg/mL.
[0288] According to another embodiment, the predetermined sTREM-1
value, preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 400
pg/mL to about 500 pg/mL.
[0289] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 400, 405, 410,
415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475,
480, 485, 490, 495 or 500 pg/mL.
[0290] According to another embodiment, the predetermined sTREM-1
value, preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 500
pg/mL to about 600 pg/mL.
[0291] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 500, 505, 510,
515, 520, 525, 530, 535, 540, 545, 550, 555, 560, 565, 570, 575,
580, 585, 590, 595 or 600 pg/mL.
[0292] According to one embodiment, the predetermined sTREM-1
value, preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, determined with an
enzyme-linked immunosorbent assay (ELISA).
[0293] As indicated above, examples of ELISA assays include,
without being limited to, the TREM-1 Quantikine ELISA kit
(reference DTRM10C from R&D Systems); the human TREM-1 DuoSet
(references DY1278B and DY1278BE from R&D Systems), the sTREM-1
ELISA (reference sTREM-1 ELISA from iQProducts).
[0294] In one embodiment, the enzyme-linked immunosorbent assay
(ELISA) is the TREM-1 Quantikine ELISA kit (reference DTRM10C from
R&D Systems).
[0295] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 50
pg/mL to about 1000 pg/mL as determined using an ELISA, or a
corresponding sTREM-1 level, preferably a blood, plasma or serum
level as determined using another immunoassay, such as an
electrochemiluminescence immunoassay (ECLIA) or an enzyme-linked
fluorescence assay (ELFA).
[0296] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 50, 75, 100,
125, 150, 175, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290,
300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420,
430, 440, 450, 460, 470, 480, 490, 500, 520, 540, 560, 580, 600,
620, 640, 660, 680, 700, 720, 740, 760, 780, 800, 820, 840, 860,
880, 900, 920, 940, 960, 980 or 1000 pg/mL, as determined using an
ELISA, or a corresponding sTREM-1 level, preferably a blood, plasma
or serum level as determined using another immunoassay, such as an
ECLIA or an ELFA.
[0297] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 300
pg/mL to about 800 pg/mL as determined using an ELISA, or a
corresponding sTREM-1 level, preferably a blood, plasma or serum
level as determined using another immunoassay, such as an ECLIA or
an ELFA.
[0298] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 300, 305, 310,
315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375,
380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440,
445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 505,
510, 515, 520, 525, 530, 535, 540, 545, 550, 555, 560, 565, 570,
575, 580, 585, 590, 595, 600, 605, 610, 615, 620, 625, 630, 635,
640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690, 695, 700,
705, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760, 765,
770, 775, 780, 785, 790, 795 or 800 pg/mL as determined using an
ELISA, or a corresponding sTREM-1 level, preferably a blood, plasma
or serum level as determined using another immunoassay, such as an
ECLIA or an ELFA.
[0299] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 350
pg/mL to about 600 pg/mL as determined using an ELISA, or a
corresponding sTREM-1 level, preferably a blood, plasma or serum
level as determined using another immunoassay, such as an ECLIA or
an ELFA.
[0300] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 350, 355, 360,
365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425,
430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490,
495, 500, 505, 510, 515, 520, 525, 530, 535, 540, 545, 550, 555,
560, 565, 570, 575, 580, 585, 590, 595 or 600 pg/mL as determined
using an ELISA, or a corresponding sTREM-1 level, preferably a
blood, plasma or serum level as determined using another
immunoassay, such as an ECLIA or an ELFA.
[0301] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 400
pg/mL to about 500 pg/mL as determined using an ELISA, or a
corresponding sTREM-1 level, preferably a blood, plasma or serum
level as determined using another immunoassay, such as an ECLIA or
an ELFA.
[0302] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 400, 405, 410,
415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475,
480, 485, 490, 495 or 500 pg/mL as determined using an ELISA, or a
corresponding sTREM-1 level, preferably a blood, plasma or serum
level as determined using another immunoassay, such as an ECLIA or
an ELFA.
[0303] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 500
pg/mL to about 600 pg/mL as determined using an ELISA, or a
corresponding sTREM-1 level, preferably a blood, plasma or serum
level as determined using another immunoassay, such as an ECLIA or
an ELFA.
[0304] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 500, 505, 510,
515, 520, 525, 530, 535, 540, 545, 550, 555, 560, 565, 570, 575,
580, 585, 590, 595 or 600 pg/mL as determined using an
enzyme-linked immunosorbent assay (ELISA), or a corresponding
sTREM-1 level, preferably a blood, plasma or serum level as
determined using another immunoassay, such as an ECLIA or an
ELFA.
[0305] According to one embodiment, the predetermined sTREM-1
value, preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, determined with an
electrochemiluminescence immunoassay (ECLIA).
[0306] As indicated above, examples of electrochemiluminescence
immunoassays (ECLIAs) include Elecsys.RTM. (Roche Diagnostics).
[0307] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 20
pg/mL to about 6000 pg/mL as determined using an
electrochemiluminescence immunoassay (ECLIA), or a corresponding
sTREM-1 level, preferably a blood, plasma or serum level as
determined using another immunoassay, such as an enzyme-linked
immunosorbent assay (ELISA) or an enzyme-linked fluorescence assay
(ELFA).
[0308] In one embodiment the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 20, 50, 100,
150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750,
800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350,
1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900,
1950, 2000, 2050, 2100, 2150, 2200, 2250, 2300, 2350, 2400, 2450,
2500, 2550, 2600, 2650, 2700, 2750, 2800, 2850, 2900, 2950, 3000,
3050, 3100, 3150, 3200, 3250, 3300, 3350, 3400, 3450, 3500, 3550,
3600, 3650, 3700, 3750, 3800, 3850, 3900, 3950, 4000, 4050, 4100,
4150, 4200, 4250, 4300, 4350, 4400, 4450, 4500, 4550, 4600, 4650,
4700, 4750, 4800, 4850, 4900, 4950, 5000, 5050, 5100, 5150, 5200,
5250, 5300, 5350, 5400, 5450, 5500, 5550, 5600, 5650, 5700, 5750,
5800, 5850, 5900, 5950 or 6000 pg/mL as determined using an ECLIA,
or a corresponding sTREM-1 level, preferably a blood, plasma or
serum level as determined using another immunoassay, such as an
ELISA or an ELFA.
[0309] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 30
pg/mL to about 3000 pg/mL as determined using an ECLIA, or a
corresponding sTREM-1 level, preferably a blood, plasma or serum
level as determined using another immunoassay, such as an ELISA or
an ELFA.
[0310] In one embodiment the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 30, 50, 100,
150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750,
800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350,
1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900,
1950, 2000, 2050, 2100, 2150, 2200, 2250, 2300, 2350, 2400, 2450,
2500, 2550, 2600, 2650, 2700, 2750, 2800, 2850, 2900, 2950 or 3000
pg/mL as determined using an ECLIA, or a corresponding sTREM-1
level, preferably a blood, plasma or serum level as determined
using another immunoassay, such as an ELISA or an ELFA.
[0311] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 500
pg/mL to about 1500 pg/mL, preferably from about 800 pg/mL to about
1200 pg/mL, as determined using an ECLIA, or a corresponding
sTREM-1 level, preferably a blood, plasma or serum level as
determined using another immunoassay, such as an ELISA or an
ELFA.
[0312] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 500
pg/mL to about 600 pg/mL, from about 600 pg/mL to about 700 pg/mL,
from about 700 pg/mL to about 800 pg/mL, from about 800 pg/mL to
about 900 pg/mL or from about 900 pg/mL to about 1000 pg/mL as
determined using an ECLIA, or a corresponding sTREM-1 level,
preferably a blood, plasma or serum level as determined using
another immunoassay, such as an ELISA or an ELFA.
[0313] In one embodiment the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 500, 520, 540,
560, 580, 600, 620, 640, 660, 680, 700, 720, 740, 760, 780, 800,
820, 840, 860, 880, 900, 920, 940, 960, 980, 1000, 1020, 1040,
1060, 1080, 1100, 1120, 1140, 1160, 1180, 1200, 1220, 1240, 1260,
1280, 1300, 1320, 1340, 1360, 1380, 1400, 1420, 1440, 1460, 1480 or
1500 pg/mL as determined using an ECLIA, or a corresponding sTREM-1
level, preferably a blood, plasma or serum level as determined
using another immunoassay, such as an ELISA or an ELFA.
[0314] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 1000
pg/mL to about 2000 pg/mL, preferably from about 1200 pg/mL to
about 1600 pg/mL, as determined using an ECLIA, or a corresponding
sTREM-1 level, preferably a blood, plasma or serum level as
determined using another immunoassay, such as an ELISA or an
ELFA.
[0315] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 1000
pg/mL to about 1100 pg/mL, from about 1100 pg/mL to about 1200
pg/mL, from about 1200 pg/mL to about 1300 pg/mL, from about 1300
pg/mL to about 1400 pg/mL, from about 1400 pg/mL to about 1500
pg/mL, from about 1500 pg/mL to about 1600 pg/mL, from about 1600
pg/mL to about 1700 pg/mL, from about 1700 pg/mL to about 1800
pg/mL, from about 1800 pg/mL to about 1900 pg/mL or from about 1900
pg/mL to about 2000 pg/mL as determined using an ECLIA, or a
corresponding sTREM-1 level, preferably a blood, plasma or serum
level as determined using another immunoassay, such as an ELISA or
an ELFA.
[0316] In one embodiment the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 1000, 1020,
1040, 1060, 1080, 1100, 1120, 1140, 1160, 1180, 1200, 1220, 1240,
1260, 1280, 1300, 1320, 1340, 1360, 1380, 1400, 1420, 1440, 1460,
1480, 1500, 1520, 1540, 1560, 1580, 1600, 1620, 1640, 1660, 1680,
1700, 1720, 1740, 1760, 1780, 1800, 1820, 1840, 1860, 1880, 1900,
1920, 1940, 1960, 1980 or 2000 pg/mL as determined using an ECLIA,
or a corresponding sTREM-1 level, preferably a blood, plasma or
serum level as determined using another immunoassay, such as an
ELISA or an ELFA.
[0317] According to one embodiment, the predetermined sTREM-1
value, preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, determined with an
enzyme-linked fluorescent assay (ELFA).
[0318] As indicated above, examples of enzyme-linked fluorescent
assays (ELFA) include VIDAS.RTM. (Biomerieux).
[0319] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 30
pg/mL to about 3000 pg/mL, preferably from about 300 pg/mL to about
2000 pg/mL, as determined using an ELFA, or a corresponding sTREM-1
level, preferably a blood, plasma or serum level as determined
using another immunoassay, such as enzyme-linked immunosorbent
assay (ELISA) or an electrochemiluminescence immunoassay
(ECLIA).
[0320] In one embodiment the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 30, 50, 100,
150, 200, 250, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525,
550, 575, 600, 625, 650, 675, 700, 725, 750, 775, 800, 825, 850,
875, 900, 925, 950, 975, 1000, 1025, 1050, 1075, 1100, 1125, 1150,
1175, 1200, 1225, 1250, 1275, 1300, 1325, 1350, 1375, 1400, 1425,
1450, 1475, 1500, 1525, 1550, 1575, 1600, 1625, 1650, 1675, 1700,
1725, 1750, 1775, 1800, 1825, 1850, 1875, 1900, 1925, 1950, 1975,
2000, 2050, 2100, 2150, 2200, 2250, 2300, 2350, 2400, 2450, 2500,
2550, 2600, 2650, 2700, 2750, 2800, 2850, 2900, 2950 or 3000 pg/mL
as determined using an ELFA, or a corresponding sTREM-1 level,
preferably a blood, plasma or serum level as determined using
another immunoassay, such as an ELISA or an ECLIA.
[0321] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 500
pg/mL to about 1500 pg/mL, preferably from about 800 pg/mL to about
1200 pg/mL, as determined using an ELFA, or a corresponding sTREM-1
level, preferably a blood, plasma or serum level as determined
using another immunoassay, such as an ELISA or an ECLIA.
[0322] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 500
pg/mL to about 600 pg/mL, from about 600 pg/mL to about 700 pg/mL,
from about 700 pg/mL to about 800 pg/mL, from about 800 pg/mL to
about 900 pg/mL or from about 900 pg/mL to about 1000 pg/mL as
determined using an ELFA, or a corresponding sTREM-1 level,
preferably a blood, plasma or serum level as determined using
another immunoassay, such as an ELISA or an ECLIA.
[0323] In one embodiment the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 500, 520, 540,
560, 580, 600, 620, 640, 660, 680, 700, 720, 740, 760, 780, 800,
820, 840, 860, 880, 900, 920, 940, 960, 980, 1000, 1020, 1040,
1060, 1080, 1100, 1120, 1140, 1160, 1180, 1200, 1220, 1240, 1260,
1280, 1300, 1320, 1340, 1360, 1380, 1400, 1420, 1440, 1460, 1480 or
1500 pg/mL as determined using an ELFA, or a corresponding sTREM-1
level, preferably a blood, plasma or serum level as determined
using another immunoassay, such as an ELISA or an ECLIA.
[0324] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 1000
pg/mL to about 2000 pg/mL, preferably from about 1200 pg/mL to
about 1600 pg/mL, as determined using an ELFA, or a corresponding
sTREM-1 level, preferably a blood, plasma or serum level as
determined using another immunoassay, such as an ELISA or an
ECLIA.
[0325] In one embodiment, the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, ranging from about 1000
pg/mL to about 1100 pg/mL, from about 1100 pg/mL to about 1200
pg/mL, from about 1200 pg/mL to about 1300 pg/mL, from about 1300
pg/mL to about 1400 pg/mL, from about 1400 pg/mL to about 1500
pg/mL, from about 1500 pg/mL to about 1600 pg/mL, from about 1600
pg/mL to about 1700 pg/mL, from about 1700 pg/mL to about 1800
pg/mL, from about 1800 pg/mL to about 1900 pg/mL or from about 1900
pg/mL to about 2000 pg/mL as determined using an ELFA, or a
corresponding sTREM-1 level, preferably a blood, plasma or serum
level as determined using another immunoassay, such as an ELISA or
an ECLIA.
[0326] In one embodiment the predetermined sTREM-1 value,
preferably the predetermined sTREM-1 value obtained from a
reference population as described hereinabove, is a sTREM-1 level,
preferably a blood, plasma or serum level, of about 1000, 1020,
1040, 1060, 1080, 1100, 1120, 1140, 1160, 1180, 1200, 1220, 1240,
1260, 1280, 1300, 1320, 1340, 1360, 1380, 1400, 1420, 1440, 1460,
1480, 1500, 1520, 1540, 1560, 1580, 1600, 1620, 1640, 1660, 1680,
1700, 1720, 1740, 1760, 1780, 1800, 1820, 1840, 1860, 1880, 1900,
1920, 1940, 1960, 1980 or 2000 pg/mL as determined using an ELFA,
or a corresponding sTREM-1 level, preferably a blood, plasma or
serum level as determined using another immunoassay, such as an
ELISA or an ECLIA.
[0327] Accordingly, in one embodiment, the present invention
relates to an in vitro method for identifying a human subject
suffering from an inflammatory disorder, preferably SIRS, sepsis or
septic shock, susceptible to respond to an administration of a
therapy, in particular of a TREM-1 inhibitor, said method
comprising: [0328] a) measuring the level of soluble Triggering
Receptors Expressed on Myeloid cells-1 (sTREM-1) in a biological
sample from the human subject; [0329] b) comparing the level of
sTREM-1 measured at step a) to a predetermined sTREM-1 value,
preferably obtained from a reference population sTREM-1, said
predetermined sTREM-1 value being a blood sTREM-1 level ranging
from about 350 pg/mL to about 600 pg/mL; [0330] c) identifying a
human subject suffering from an inflammatory disorder, preferably
SIRS, sepsis or septic shock, with a level of sTREM-1 measured at
step a) higher than the predetermined sTREM-1 value of step b) as
susceptible to respond to an administration of a therapy, in
particular of a TREM-1 inhibitor.
[0331] In another embodiment, the present invention relates to an
in vitro method for identifying a human subject suffering from an
inflammatory disorder, preferably SIRS, sepsis or septic shock,
susceptible to respond to an administration of a therapy, in
particular of a TREM-1 inhibitor, said method comprising: [0332] a)
measuring the level of soluble Triggering Receptors Expressed on
Myeloid cells-1 (sTREM-1) in a biological sample from the human
subject; [0333] b) comparing the level of sTREM-1 measured at step
a) to a predetermined sTREM-1 value, preferably obtained from a
reference population sTREM-1, said predetermined sTREM-1 value
being a blood sTREM-1 level ranging from about 300 pg/mL to about
800 pg/mL, preferably from about 350 pg/mL to about 600 pg/mL, as
determined using an enzyme-linked immunosorbent assay (ELISA), or a
corresponding blood sTREM-1 level as determined using another
immunoassay, such as an ECLIA or an ELFA; [0334] c) identifying a
human subject suffering from an inflammatory disorder, preferably
SIRS, sepsis or septic shock, with a level of sTREM-1 measured at
step a) higher than the predetermined sTREM-1 value of step b) as
susceptible to respond to an administration of a therapy, in
particular of a TREM-1 inhibitor.
[0335] In another embodiment, the present invention relates to an
in vitro method for identifying a human subject suffering from an
inflammatory disorder, preferably SIRS, sepsis or septic shock,
susceptible to respond to an administration of a therapy, in
particular of a TREM-1 inhibitor, said method comprising: [0336] a)
measuring the level of soluble Triggering Receptors Expressed on
Myeloid cells-1 (sTREM-1) in a biological sample from the human
subject; [0337] b) comparing the level of sTREM-1 measured at step
a) to a predetermined sTREM-1 value, preferably obtained from a
reference population sTREM-1, said predetermined sTREM-1 value
being a blood sTREM-1 level ranging from about 20 pg/mL to about
6000 pg/mL, preferably from about 30 pg/mL to about 3000 pg/mL, as
determined using an electrochemiluminescence immunoassay (ECLIA),
or a corresponding blood sTREM-1 level as determined using another
immunoassay, such as an ELISA or an ELFA; [0338] c) identifying a
human subject suffering from an inflammatory disorder, preferably
SIRS, sepsis or septic shock, with a level of sTREM-1 measured at
step a) higher than the predetermined sTREM-1 value of step b) as
susceptible to respond to an administration of a therapy, in
particular of a TREM-1 inhibitor.
[0339] In another embodiment, the present invention relates to an
in vitro method for identifying a human subject suffering from an
inflammatory disorder, preferably SIRS, sepsis or septic shock,
susceptible to respond to an administration of a therapy, in
particular of a TREM-1 inhibitor, said method comprising: [0340] a)
measuring the level of soluble Triggering Receptors Expressed on
Myeloid cells-1 (sTREM-1) in a biological sample from the human
subject; [0341] b) comparing the level of sTREM-1 measured at step
a) to a predetermined sTREM-1 value, preferably obtained from a
reference population sTREM-1, said predetermined sTREM-1 value
being a blood sTREM-1 level ranging from about 30 pg/mL to about
3000 pg/mL, preferably from about 300 pg/mL to about 2000 pg/mL, as
determined using an enzyme-linked fluorescent assay (ELFA), or a
corresponding blood sTREM-1 level as determined using another
immunoassay, such as an ELISA or an ECLIA; [0342] c) identifying a
human subject suffering from an inflammatory disorder, preferably
SIRS, sepsis or septic shock, with a level of sTREM-1 measured at
step a) higher than the predetermined sTREM-1 value of step b) as
susceptible to respond to an administration of a therapy, in
particular of a TREM-1 inhibitor.
[0343] According to one embodiment, the therapy is an
immunomodulatory therapy or an anti-inflammatory therapy.
[0344] Examples of immunomodulatory therapies or anti-inflammatory
therapies include, without being limited to, checkpoint inhibitors
such as anti-PD-1, anti-PD-L1 and anti-CTLA4; TLR (Toll-like
receptors) inhibitors; cytokine inhibitors such as anti-cytokine or
anti-cytokine receptors (for example IL-1RA for interleukin-1
receptor antagonist); G-CSF (granulocyte-colony stimulating
factor); GM-CSF (granulocyte-macrophage colony-stimulating factor);
IL-7 (interleukin-7); inhibitors of immunostimulants such as CD28
antagonist peptides and antibodies, in particular monoclonal
antibodies, against CD28; and cellular therapies such as adoptive
cell therapies.
[0345] In one embodiment, the therapy is selected from the group
consisting of checkpoint inhibitors; TLR (Toll-like receptors)
inhibitors; cytokine inhibitors including anti-cytokine or
anti-cytokine receptors; G-CSF (granulocyte-colony stimulating
factor); IL-7 (interleukin-7); inhibitors of immunostimulants; and
cellular therapies. In other words, in one embodiment, the therapy
comprises or consists in the administration of at least one of a
checkpoint inhibitor; a TLR (Toll-like receptors) inhibitor; a
cytokine inhibitor; G-CSF (granulocyte-colony stimulating factor);
IL-7 (interleukin-7); GM-CSF (granulocyte-macrophage
colony-stimulating factor); an inhibitor of immunostimulants; or a
cellular therapy.
[0346] As used herein, checkpoint inhibitors (CPI, that may also be
referred to as immune checkpoint inhibitors or ICI) refer to
compounds that block the interactions between inhibitory receptors
expressed on T cells and their ligands. Checkpoint inhibitors
include antibodies, in particular monoclonal antibodies, and
non-antibody inhibitors such as small molecule inhibitors.
[0347] Examples of checkpoint inhibitors include, without being
limited to, inhibitors of the cell surface receptor PD-1
(programmed cell death protein 1), also known as CD279 (cluster
differentiation 279); inhibitors of the ligand PD-L1 (programmed
death-ligand 1), also known as CD274 (cluster of differentiation
274) or B7-H1 (B7 homolog 1); inhibitors of the cell surface
receptor CTLA4 or CTLA-4 (cytotoxic T-lymphocyte-associated protein
4), also known as CD152 (cluster of differentiation 152);
inhibitors of LAG-3 (lymphocyte-activation gene 3), also known as
CD223 (cluster differentiation 223); inhibitors of TIM-3 (T-cell
immunoglobulin and mucin-domain containing-3), also known as HAVCR2
(hepatitis A virus cellular receptor 2) or CD366 (cluster
differentiation 366); inhibitors of TIGIT (T cell immunoreceptor
with Ig and ITIM domains), also known as VSIG9 (V-Set And
Immunoglobulin Domain-Containing Protein 9) or VSTM3 (V-Set And
Transmembrane Domain-Containing Protein 3); inhibitors of BTLA (B
and T lymphocyte attenuator), also known as CD272 (cluster
differentiation 272); inhibitors of CEACAM-1 (carcinoembryonic
antigen-related cell adhesion molecule 1) also known as CD66a
(cluster differentiation 66a).
[0348] As used herein, TLR inhibitors (sometimes also referred to
as TLR antagonists) refer to compounds that block TLR signaling.
TLR inhibitors may act by blocking the binding of TLR ligands to
the receptor or by blocking the intracellular signaling pathways to
stop the signal transduction (Gao et al., Front Physiol. 2017; 8:
508). TLR inhibitors include, without being limited to, small
molecule inhibitors; antibodies, in particular monoclonal
antibodies; oligonucleotides; lipid-A analogs; microRNAs; and
nano-inhibitors.
[0349] As used herein, cytokine inhibitors refer to compound that
decrease the synthesis of cytokines, decrease their concentration
in free active form, block their interaction with specific
receptors or interfere with the signaling of cytokine receptors.
Cytokine inhibitors include, without being limited to, antibodies,
in particular monoclonal antibodies against cytokines (i.e.,
anti-cytokine antibodies); antibodies, in particular monoclonal
antibodies against cytokine receptors (i.e., anti-cytokine receptor
antibodies); cytokine receptor antagonists; and soluble receptors
acting as decoy receptors.
[0350] According to one embodiment, the therapy is a therapy
seeking to inhibit or remove endotoxins, also referred to herein as
anti-endotoxin therapy.
[0351] Examples of anti-endotoxin therapies include, without being
limited to, endotoxin inhibitors such as anti-endotoxin antibodies,
in particular anti-endotoxin monoclonal antibodies; recombinant
alkaline phosphatase, such as, for example, human recombinant AP
(recAP); and hemoperfusion which allows for the removal of a toxin
by direct contact of the blood in an extracorporeal circulation
with a material that adsorbs the toxin, such as, for example,
polymyxin B immobilized fiber cartridge (PMX-DHP).
[0352] According to one embodiment, the therapy is a vasopressor
therapy. In other words, according to one embodiment, the therapy
comprises or consists in the administration of a vasopressor.
[0353] Examples of vasopressor therapies include, without
limitation, the administration of vasoactive catecholamine hormones
such as norepinephrine, dopamine, epinephrine; vasopressin; and/or
phenylephrine.
[0354] According to one embodiment, the therapy is an angiogenesis
inhibitor, in particular an angiopoietin-2 (Ang-2 or Ang2)
inhibitor. In other words, according to one embodiment, the therapy
comprises or consists in the administration of an angiogenesis
inhibitor, in particular an angiopoietin-2 (Ang-2 or Ang2)
inhibitor.
[0355] The angiopoietin-Tie signaling system, comprising notably
angiopoetin-1 (Ang-1) and Tie 2, is a vascular-specific receptor
tyrosine kinase pathway that is essential for normal vascular
development. Initially Ang-2 was identified as an antagonist of
Ang-1, inhibiting the activation of Tie2 by Ang-1. However, it
remains unclear whether Ang-2 is an antagonist or agonist of Tie2
in settings of vascular remodeling. A number of studies indicate
that Ang-2 plays a role in inflammation-induced vascular remodeling
and in tumor angiogenesis and growth (Thurston & Daly, Cold
Spring Harb Perspect Med. 2012 Sep. 1; 2(9):a006550).
[0356] As used herein, Ang-2 inhibitors (sometimes also referred to
as Ang2 antagonists) refer to compounds that block Ang-2 signaling,
notably that block Ang-2 and Tie2 interaction. According to the
invention, Ang-2 inhibitors thus include inhibitors binding Ang-2
and inhibitors binding Tie2. Examples of Ang-2 inhibitors include,
without being limited to antibodies, in particular monoclonal
antibodies, against Ang-2; Ang2-Blocking Antibody also referred to
as ABA (Han et al., Sci Transl Med. 2016 Apr. 20; 8(335):335ra55);
Ang2-Binding and Tie2-Activating Antibody also referred to as ABTAA
(Han et al., Sci Transl Med. 2016 Apr. 20; 8(335):335ra55);
nanobodies against Ang-2, including bispecific nanobodies that
inhibit for example VEGF (vascular endothelial growth factor) and
Ang-2; peptibodies against Ang-2, corresponding to the fusion of an
inhibitory Ang-2 binding peptide and an Ig Fc domain; soluble decoy
receptors also referred to as soluble ligand-trap receptors;
oligonucleotides such as antisense RNAs; aptamers such as RNA
aptamers specifically blocking Ang-2.
[0357] According to one embodiment, the therapy is an
adrenomedullin (ADM) and adrenomedullin-targeted therapy.
[0358] Examples of adrenomedullin and adrenomedullin-targeted
therapies include, without being limited to, adrenomedullin (ADM)
such as, for example, a bolus of ADM or a continuous infusion of
ADM; antibodies recognizing ADM (anti-AMD antibodies) including
antibodies recognizing the biologically active ADM (anti-bio-ADM
antibodies), in particular monoclonal antibodies including
non-neutralizing antibodies such as Adrecizumab.
[0359] According to one embodiment, the therapy is a TREM-1
inhibitor. In other words, according to one embodiment, the therapy
comprises or consists in the administration of a TREM-1
inhibitor.
[0360] According to the present invention, a TREM-1 inhibitor is an
active agent able to inhibit TREM-1 function, activity or
expression.
[0361] In one embodiment, the TREM-1 inhibitor is selected from the
group consisting of peptides inhibiting the function, activity or
expression of TREM-1; antibodies directed to TREM-1 and/or sTREM-1,
or TREM-1 and/or sTREM-1 ligand; small molecules inhibiting the
function, activity or expression of TREM-1; siRNAs directed to
TREM-1; shRNAs directed to TREM-1; antisense oligonucleotides
directed to TREM-1; ribozymes directed to TREM-1 and aptamers
directed to TREM-1.
[0362] Accordingly, in one embodiment, the present invention
relates to an in vitro method for identifying a human subject
suffering from an inflammatory disorder, preferably SIRS, sepsis or
septic shock, susceptible to respond to an administration of a
TREM-1 inhibitor as described hereinabove, said method comprising:
[0363] a) measuring the level of soluble Triggering Receptors
Expressed on Myeloid cells-1 (sTREM-1) in a biological sample from
the human subject; [0364] b) comparing the level of sTREM-1
measured at step a) to a predetermined sTREM-1 value as described
hereinabove; [0365] c) identifying a human subject suffering from
an inflammatory disorder, preferably SIRS, sepsis or septic shock
with a level of sTREM-1 measured at step a) higher than the
predetermined sTREM-1 value of step b) as susceptible to respond to
an administration of a TREM-1 inhibitor as described
hereinabove.
[0366] Examples of peptides inhibiting the function, activity or
expression of TREM-1 include, without being limited to, peptides
targeting TREM-1 ligand, such as, for example, TLT-1 peptides.
[0367] In one embodiment, the TREM-1 inhibitor is a peptide
inhibiting TREM-1 through its binding to TREM-1 ligand.
[0368] In one embodiment, the TREM-1 inhibitor is a TLT-1
peptide.
[0369] In one embodiment, the TREM-1 inhibitor is a short TLT-1
peptide consisting of between 6 and 20 consecutive amino acids from
the human TLT-1 having an amino acid sequence as set forth in SEQ
ID NO: 7 (MGLTLLLLLLLGLEGQGIVGSLPEVLQAPVG
SSILVQCHYRLQDVKAQKVWCRFLPEGCQPLVSSAVDRRAPAGRRTFLTDLGG
GLLQVEMVTLQEEDAGEYGCMVDGARGPQILHRVSLNILPPEEEEETHKIGSLA
ENAFSDPAGSANPLEPSQDEKSIPLIWGAVLLVGLLVAAVVLFAVMAKRKQGN
RLGVCGRFLSSRVSGMNPSSVVHHVSDSGPAAELPLDVPHIRLDSPPSFDNTTYT
SLPLDSPSGKPSLPAPSSLPPLPPKVLVCSKPVTYATVIFPGGNKGGGTSCGPAQ NPPNNQTPSS)
or a sequence having at least 60, 65, 70, 75, 80, 85 or 90%
identity with the amino acid sequence as set forth in SEQ ID NO:
7.
[0370] In one embodiment, the TREM-1 inhibitor is a short TLT-1
peptide consisting of between 6 and 20 consecutive amino acids from
the human TLT-1 having an amino acid sequence as set forth in SEQ
ID NO: 7 or a function-conservative variant or derivative
thereof.
[0371] In one embodiment, the TREM-1 inhibitor is a TLT-1 peptide
consisting of 6 to 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids
and comprising an amino acid sequence as set forth in SEQ ID NO: 8
(LQEEDAGEYGCMVDGAR) also referred to as LR17, SEQ ID NO: 9
(LQEEDAGEYGCM) also referred to as LR12, SEQ ID NO: 10 (LQEEDA)
also referred to as LR6-1, SEQ ID NO: 11 (EDAGEY) also referred to
as LR6-2, or SEQ ID NO: 12 (GEYGCM) also referred to as LR6-3, or a
sequence having at least 60, 65, 70, 75, 80, 85 or 90% identity
with the amino acid sequence as set forth in SEQ ID NO: 8, SEQ ID
NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, or SEQ ID NO: 12,
respectively.
[0372] In one embodiment, the TREM-1 inhibitor is a TLT-1 peptide
consisting of 6 to 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids
and comprising an amino acid sequence as set forth in SEQ ID NO: 8,
SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, or SEQ ID NO: 12 or a
function-conservative variant or derivative thereof.
[0373] In one embodiment, the TREM-1 inhibitor is a TLT-1 peptide
comprising or consisting of an amino acid sequence as set forth in
SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, or SEQ ID
NO: 12 or a sequence having at least 60, 65, 70, 75, 80, 85 or 90%
identity with the amino acid sequence as set forth in SEQ ID NO: 8,
SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, or SEQ ID NO: 12,
respectively.
[0374] In one embodiment, the TREM-1 inhibitor is a TLT-1 peptide
comprising or consisting of an amino acid sequence as set forth in
SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, or SEQ ID
NO: 12 or a function-conservative variant or derivative
thereof.
[0375] In one embodiment, the TREM-1 inhibitor is a TLT-1 peptide
having an amino acid sequence as set forth in SEQ ID NO: 8, SEQ ID
NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, or SEQ ID NO: 12 or a sequence
having at least 60, 65, 70, 75, 80, 85 or 90% identity with the
amino acid sequence as set forth in SEQ ID NO: 8, SEQ ID NO: 9, SEQ
ID NO: 10, SEQ ID NO: 11, or SEQ ID NO: 12, respectively.
[0376] In one embodiment, the TREM-1 inhibitor is a TLT-1 peptide
having an amino acid sequence as set forth in SEQ ID NO: 8, SEQ ID
NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, or SEQ ID NO: 12 or a
function-conservative variant or derivative thereof.
[0377] In one embodiment, the TREM-1 inhibitor is a TLT-1 peptide
having an amino acid sequence as set forth in SEQ ID NO: 9, also
known as LR12 or nangibotide or motrem, or a sequence having at
least 60, 65, 70, 75, 80, 85 or 90% identity with the amino acid
sequence as set forth in SEQ ID NO: 9.
[0378] In one embodiment, the TREM-1 inhibitor is a TLT-1 peptide
having an amino acid sequence as set forth in SEQ ID NO: 9, also
known as LR12 or nangibotide or motrem, or a function-conservative
variant or derivative thereof.
[0379] As used herein, the term "function-conservative variants"
denotes peptides derived from the TLT-1 peptides as described
hereinabove, in which a given amino acid residue in a peptide has
been changed without altering the overall conformation and function
of said TLT-1 peptides, including, but not limited to, replacement
of an amino acid with one having similar properties (such as, for
example, similar polarity, similar hydrogen bonding potential,
acidic or basic amino acid replaced by another acidic or basic
amino acid, hydrophobic amino acid replaced by another hydrophobic
amino acid, aromatic amino acid replaced by another aromatic amino
acid).
[0380] It is commonly known that amino acids other than those
indicated as conserved may differ in a protein so that the percent
of protein or amino acid sequence similarity between any two
proteins of similar function may vary and may be, for example, from
70% to 99% as determined according to an alignment method such as
by the Cluster Method, wherein similarity is based on the MEGALIGN
algorithm.
[0381] A "function-conservative variant" also includes TLT-1
peptides which have at least 20%, 30%, 40%, 50%, or 60% amino acid
identity with the TLT-1 peptides as described hereinabove, for
example as determined by BLAST or FASTA algorithms, and which have
the same or substantially similar properties or functions as the as
the TLT-1 peptides as described hereinabove. Preferably
"function-conservative variants" include TLT-1 peptides which have
at least 60%, 65%, 70%, 75%, 80%, 85% or 90% amino acid identity
with the TLT-1 peptides as described hereinabove and which have the
same or substantially similar properties or functions as the TLT-1
peptides as described hereinabove.
[0382] As used herein, the term "derivative" refers to a variation
of a peptide or of a function-conservative variant thereof that is
otherwise modified in order to alter the in vitro or in vivo
conformation, activity, specificity, efficacy or stability of the
peptide. For example, said variation may encompass modification by
covalent attachment of any type of molecule to the peptide or by
addition of chemical compound(s) to any of the amino-acids of the
peptide.
[0383] In one embodiment, the TLT-1 peptide or
function-conservative variants or derivatives thereof as described
hereinabove may have D- or L-configuration.
[0384] In one embodiment, the amino acid from the amino end of the
TLT-1 peptide or function-conservative variant or derivative
thereof as described hereinabove has an acetylated terminal amino
group, and the amino acid from the carboxyl end has an amidated
terminal carboxy group.
[0385] In addition, the TLT-1 peptide or function-conservative
variant or derivative thereof as described hereinabove may undergo
reversible chemical modifications in order to increase its
bioavailability (including stability and fat solubility) and its
ability to pass the blood-brain barrier and epithelial tissue.
Examples of such reversible chemical modifications include
esterification of the carboxy groups of glutamic and aspartic amino
acids with an alcohol, thereby removing the negative charge of the
amino acid and increasing its hydrophobicity. This esterification
is reversible, as the ester link formed is recognized by
intracellular esterases which hydrolyze it, restoring the charge to
the aspartic and glutamic residues. The net effect is an
accumulation of intracellular peptide, as the internalized,
de-esterified peptide cannot cross the cell membrane.
[0386] Another example of such reversible chemical modifications
includes the addition of a further peptide sequence, which allows
the increase of the membrane permeability, such as a TAT peptide or
Penetratin peptide (see--Charge-Dependent Translocation of the
Trojan. A Molecular View on the Interaction of the Trojan Peptide
Penetratin with the 15 Polar Interface of Lipid Bilayers.
Biophysical Journal, Volume 87, Issue 1, 1 Jul. 2004, Pages
332-343).
[0387] The TLT-1 peptides or function-conservative variants or
derivatives thereof as described hereinabove may be obtained
through conventional methods of solid-phase chemical peptide
synthesis, following Fmoc and/or Boc-based methodology (see
Pennington, M. W. and Dunn, B. N. (1994). Peptide synthesis
protocols. Humana Press, Totowa.).
[0388] Alternatively, the TLT-1 peptides or function-conservative
variants or derivatives as described hereinabove may be obtained
through conventional methods based on recombinant DNA technology,
e.g., through a method that, in brief, includes inserting the
nucleic acid sequence coding for the peptide into an appropriate
plasmid or vector, transforming competent cells for said plasmid or
vector, and growing said cells under conditions that allow the
expression of the peptide and, if desired, isolating and
(optionally) purifying the peptide through conventional means known
to experts in these matters or eukaryotic cells that express the
peptide. A review of the principles of recombinant DNA technology
may be found, for example, in the text book entitled "Principles of
Gene Manipulation: An Introduction to Genetic Engineering," R. W.
Old & S. B. Primrose, published by Blackwell Scientific
Publications, 4th Edition (1989).
[0389] According to one embodiment, the TREM-1 inhibitor is a TLT-1
peptide as described hereinabove, in particular a TLT-1 peptide
having an amino acid sequence as set forth in SEQ ID NO: 9, also
known as LR12 or nangibotide or motrem, or a sequence having at
least 60, 65, 70, 75, 80, 85 or 90% identity with the amino acid
sequence as set forth in SEQ ID NO: 9, and said TLT-1 peptide is to
be administered to the human subject by continuous infusion,
preferably by continuous intravenous infusion, at a dose ranging
from about 0.1 mg per kg bodyweight per hour (mg/kg/h) to about 2.5
mg/kg/h, preferably from about 0.3 mg/kg/h to about 1 mg/kg/h, even
more preferably from about 0.3 mg/kg/h to about 0.9 mg/kg/h. In one
embodiment, said TLT-1 peptide is to be administered to the human
subject by continuous infusion, preferably by continuous
intravenous infusion, at a dose ranging from about 0.15 g/24h to
about 4.5 g/24h, preferably from about 0.5 g/24h to about 2 g/24h,
even more preferably from about 0.5 g/24h to about 1.5 g/24h.
[0390] In one embodiment, said TLT-1 peptide is to be administered
to the human subject by continuous infusion, preferably by
continuous intravenous infusion, at a dose of about 0.1, 0.2, 0.3,
0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or 1 mg/kg/h, preferably at a dose of
about 0.3 mg/kg/h. In one embodiment, said TLT-1 peptide is to be
administered to the human subject by continuous infusion,
preferably by continuous intravenous infusion, at a dose of about
0.3 mg/kg/h or about 1 mg/kg/h. In one embodiment, said TLT-1
peptide is to be administered to the human subject by continuous
infusion, preferably by continuous intravenous infusion, at a dose
of about 0.15, 0.30, 0.45, 0.60, 0.75, 0.90, 1.05, 1.2, 1.35, 1.5,
1.65, 1.8 or 2 g/24h.
[0391] In one embodiment, said TLT-1 peptide is to be administered
to the human subject for at least 24 hours and/or for at most 5
days. In one embodiment, said TLT-1 peptide is to be administered
to the human subject for 24 hours, 48 hours, 72 hours, 96 hours or
120 hours. Thus, in one embodiment, said TLT-1 peptide is to be
administered to the human subject for 1 day, 2 days, 3 days, 4 days
or 5 days.
[0392] In one embodiment, the continuous administration of a TLT-1
peptide as described hereinabove is to be preceded by the
administration of a loading dose of TLT-1 peptide. In one
embodiment, the loading dose of TLT-1 peptide is a dose ranging
from about 0.5 mg/kg to about 5 mg/kg. In one embodiment, the
loading dose of TLT-1 is a dose of about 0.5, 0.75, 1, 1.25, 1.5,
1.665, 1.75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5,
4.75 or 5 mg/kg, preferably a dose of about 1.665 mg/kg or 5 mg/kg.
In one embodiment, the loading dose of TLT-1 peptide as described
hereinabove is to be administered over about 15 min, preferably by
intravenous injection. Thus, in one embodiment, the loading dose of
TLT-1 peptide is a dose ranging from about 2 mg/kg/h to about 20
mg/kg/h. In one embodiment, the loading dose of TLT-1 peptide is a
dose of about 2, 3, 4, 5, 6, 6.66, 7, 8, 9, 10, 11, 12, 13, 14, 15,
16, 17, 18, 19 or 20 mg/kg/h, preferably at a dose of 6.66 or 20
mg/kg/h.
[0393] According to one embodiment, the therapy as described
hereinabove is to be administered between the first 2 hours and the
first 24 hours following the start of patient care, in particular
following the start of vasopressor therapy.
[0394] In one embodiment, the therapy as described hereinabove is
to be administered within the first 2, 3, 6, 9, 12, 15, 18, 21 or
24 hours following the start of patient care, in particular
following the start of vasopressor therapy.
[0395] In one embodiment, the subject is a human patient who is
awaiting the receipt of, or is receiving, medical care or
was/is/will be the subject of a medical procedure or is monitored
for the development or progression of a disease, such as an
inflammatory disorder, preferably SIRS, sepsis or septic shock.
[0396] In one embodiment, the subject is a human patient who is
hospitalized with septic shock.
[0397] In one embodiment, the subject is suffering from a
documented or suspected infection.
[0398] In one embodiment, the subject is suffering from a
community-acquired infection. In one embodiment, the subject is
suffering from a hospital-acquired infection.
[0399] Examples of infections include, without being limited to,
respiratory infections, abdominal infections and urinary tract
infections (UTIs) meningitis, endocarditis, skin infections, bone
infections, wound infections, catheter-related bloodstream
infections, device-associated infections.
[0400] In one embodiment, the subject is a man. In another
embodiment, the subject is a woman. In one embodiment, the subject
is a pregnant woman
[0401] In one embodiment, the subject is an adult. In one
embodiment, the subject is an elderly subject. In one embodiment,
the subject is a child. In one embodiment, the subject is an
infant. In one embodiment, the subject is a newborn.
[0402] In one embodiment, the subject is not immunocompromised,
immunosuppressed or immunodeficient. In another embodiment, the
subject is immunocompromised, immunosuppressed or immunodeficient.
In another embodiment, the subject is receiving an
immunosuppressive therapy.
[0403] In one embodiment, the subject is afflicted with a chronic
medical condition such as atrial fibrillation, cancer, chronic
kidney disease, chronic lung disease, cirrhosis, coronary artery
disease, deep vein thrombosis, diabetes, dyslipidemia,
endocarditis, hypertension, influenza, malaria or any other
protozoan parasitic disease, myocardial infarction, neurological
disease, non-alcoholic fatty liver disease (NAFLD), non-alcoholic
steatohepatitis (NASH), peripheral artery disease, pulmonary
fibrosis, severe obesity and stroke.
[0404] According to one embodiment, the inflammatory disorder is
selected from the group comprising or consisting of systemic
inflammatory response syndrome (SIRS), sepsis, septic shock,
sepsis-associated organ dysfunction, acute respiratory distress
syndrome (ARDS), severe acute respiratory syndrome (SARS), acute
kidney injury (AKI), pancreatitis, inflammatory bowel disease,
pneumonia, endotoxemia and hemorrhagic shock.
[0405] According to one embodiment, the inflammatory disorder is an
acute inflammatory disorder.
[0406] Examples of acute inflammatory disorders include, without
being limited to, systemic inflammatory response syndrome (SIRS),
sepsis, septic shock, acute respiratory distress syndrome (ARDS),
acute kidney injury (AKI), pancreatitis and hemorrhagic shock.
[0407] In one embodiment, the acute inflammatory disorder is
selected from the group comprising or consisting of systemic
inflammatory response syndrome (SIRS), sepsis, septic shock, acute
respiratory distress syndrome (ARDS), acute kidney injury (AKI),
pancreatitis and hemorrhagic shock.
[0408] According to one embodiment, the inflammatory disorder is
selected from the group comprising or consisting of systemic
inflammatory response syndrome (SIRS), sepsis, septic shock, acute
respiratory distress syndrome (ARDS) and acute kidney injury
(AKI).
[0409] In one embodiment, the inflammatory disorder is systemic
inflammatory response syndrome (SIRS), sepsis or septic shock.
[0410] In one embodiment, the inflammatory disorder is septic
shock.
[0411] Systemic inflammatory response syndrome (SIRS) is
characterized by systemic inflammation and widespread tissue
injury. Clinically, SIRS is defined as fulfilling at least two of
the following four criteria: fever >38.0.degree. C. or
hypothermia <36.0.degree. C.; tachycardia >90 beats/minute;
tachypnea >20 breaths/minute; and leukocytosis
>12.times.10.sup.9/L or leucopenia <4.times.10.sup.9/L (Bone
et al., Chest. 1992 June; 101(6):1644-55). SIRS may occur as a
response to a nonspecific insult of either infectious or
non-infectious origin. Examples of insults of non-infectious origin
include, without being limited to, trauma, thermal injury,
pancreatitis, autoimmune disorders and surgery. Examples of insults
of infectious origin include bacterial infections (e.g.,
respiratory infections, abdominal infections and urinary tract
infections (UTIs), fungal infections (e.g., respiratory infections)
and viral infections (e.g., respiratory infections).
[0412] As mentioned hereinabove, sepsis is defined as a
life-threatening organ dysfunction caused by a dysregulated
response of the human subject to infection (Singer et al., JAMA.
2016 Feb. 23; 315(8):801-10). Patients with sepsis can be
clinically identified as patients suffering from a documented or
suspected infection and from an organ dysfunction. In one
embodiment, organ dysfunction in a human subject can be identified
using an organ dysfunction score, i.e., a score used to assess
organ dysfunction in a human subject, notably upon admission in ICU
or emergency unit. Examples of organ dysfunction scores include,
without being limited to, the SOFA score, the qSOFA score, the MODS
(Multiple Organ Dysfunction Score), the P-MODS (Pediatric Multiple
Organ Dysfunction Score) and the LODS (Logistic Organ Dysfunction
System). In one embodiment, organ dysfunction in a human subject
can be identified as an acute change in the total SOFA score of 2
points or greater, consequent to the infection (Singer et al.,
JAMA. 2016 Feb. 23; 315(8):801-10). The baseline SOFA score can be
assumed to be zero in patients not known to have preexisting organ
dysfunction (Singer et al., JAMA. 2016 Feb. 23; 315(8):801-10).
Septic shock is defined as a subset of sepsis, in which
particularly profound circulatory, cellular, and metabolic
abnormalities are associated with a greater risk of mortality than
with sepsis alone. Thus, sepsis encompasses septic shock. Patients
with septic shock can be clinically identified as patients
suffering from sepsis and having (i) persisting hypotension
requiring vasopressors to maintain their mean arterial pressure
>65 mm Hg despite adequate volume resuscitation and (ii) a serum
lactate level >2 mmol/L (18 mg/dL) (Sepsis-3 definition as
described in Singer et al., JAMA. 2016 Feb. 23; 315(8):801-10).
With these criteria, hospital mortality is in excess of 40%.
(Singer et al., JAMA. 2016 Feb. 23; 315(8):801-10).
[0413] In one embodiment, the infection inducing the dysregulated
response of the human subject suffering from SIRS, sepsis or septic
shock is a bacterial, fungal or viral infection. Examples of
infections include, without being limited to, respiratory
infections, abdominal infections and urinary tract infections
(UTIs).
[0414] According to one embodiment, the human subject suffering
from SIRS, sepsis or septic shock may be assessed using a severity
score, i.e., a score used to assess the severity of the disease
and/or the prognosis upon admission in ICU or emergency unit.
Examples of severity scores include, without being limited to, the
APACHE II score, the APACHE III score, the APACHE IV score, the
SAPS score, the SAPS II score and the SAPS 3 score.
[0415] In one embodiment, the human subject suffering from SIRS,
sepsis or septic shock may be assessed using the APACHE II
(referring to "Acute Physiology And Chronic Health Evaluation II")
scoring system. APACHE II is commonly used to assess the severity
of disease in adult patients admitted in intensive care units and
determine their prognostic. APACHE II uses a point score ranging
from 0 to 71 that is based upon the initial values of 12 routine
physiologic measurements, age, and previous health status to
provide a general measure of severity of disease (Knaus et al.,
Crit Care Med. 1985 October; 13(10):818-29). The APACHE II score
may be determined during the first 24h upon admission in intensive
care unit (ICU) or emergency unit. A higher score is associated
with a higher predicted mortality, with a score of 25 representing
a predicted mortality of 50% and a score of over 35 representing a
predicted mortality of 80%.
[0416] In one embodiment, the human subject suffering from SIRS,
sepsis or septic shock has an APACHE II score lower than 34. In one
embodiment, the human subject suffering from SIRS, sepsis or septic
shock is not moribund.
[0417] In one embodiment, the human subject suffering from SIRS,
sepsis or septic shock may be assessed using the APACHE III
(referring to "Acute Physiology And Chronic Health Evaluation" III)
scoring system. APACHE III was redefined from the APACHE II scoring
system in order to more accurately predict hospital mortality risk
for critically ill hospitalized adults (Knaus et al., Chest. 1991
December; 100(6):1619-36).
[0418] In one embodiment, the human subject suffering from SIRS,
sepsis or septic shock may be assessed using the APACHE IV
(referring to "Acute Physiology And Chronic Health Evaluation IV")
scoring system. APACHE IV is an improved and updated model for
estimating the risk of short-term mortality as well as predicting
the length of intensive care unit (ICU) stay (Zimmerman et al.,
Crit Care Med. 2006 May; 34(5):1297-310). In the APACHE IV scoring
system, a greater number of variables are considered, notably
mechanical ventilation, thrombolysis, impact of sedation on Glasgow
Coma Scale, rescaled Glasgow Coma Scale, PaO.sub.2/FiO.sub.2 ratio
and disease-specific subgroups. APACHE IV uses a point score
ranging from 0 to 286.
[0419] In one embodiment, the human subject suffering from SIRS,
sepsis or septic shock may be assessed using the SAPS II (referring
to "Simplified Acute Physiology Score II") scoring system. SAPS II
is a scoring system for estimating in-hospital mortality in adult
patients admitted to the intensive care unit (ICU). SAPS II include
17 variables: 12 physiology variables, age, type of admission, and
3 variables regarding underlying diseases (Le Gall et al., JAMA.
1993 Dec. 22-29; 270(24):2957-63). SAPS II uses a point score
ranging from 0 to 163.
[0420] In one embodiment, the human subject suffering from SIRS,
sepsis or septic shock may be assessed using the SAPS 3 (referring
to "Simplified Acute Physiology Score III") scoring system. SAPS 3
is a scoring system for predicting hospital mortality of patients
admitted to the intensive care unit (ICU) (Metnitz et al.,
Intensive Care Med. 2005 October; 31(10): 1336-1344 and Moreno et
al., Intensive Care Med. 2005 October; 31(10):1345-55). SAPS 3 is
based on 20 different variables.
[0421] As mentioned hereinabove, the presence of organ dysfunction
associated with sepsis or septic shock may be assessed using an
organ dysfunction score such as the SOFA score, the qSOFA score,
the MODS (Multiple Organ Dysfunction Score), the P-MODS (Pediatric
Multiple Organ Dysfunction Score) or the LODS (Logistic Organ
Dysfunction System).
[0422] The Sequential Organ Failure Assessment (SOFA) score
(originally referred to as the Sepsis-related Organ Failure
Assessment, Vincent et al., Intensive Care Med. 1996 July;
22(7):707-10) is a scoring system commonly used to assess organ
dysfunction in a human subject notably upon admission in ICU or
emergency unit. The SOFA scoring system (Vincent et al., Crit Care
Med. 1998 November; 26(11):1793-800) relies on the assessment of
the respiratory system (i.e., PaO.sub.2/FiO.sub.2 (mmHg)), the
nervous system (i.e., Glasgow coma scale), the cardiovascular
system (i.e., mean arterial pressure or administration of
vasopressors required), the liver function (i.e., bilirubin (mg/dL
or .mu.mon)), coagulation (i.e., platelet count), the kidney
function (i.e., creatinine (mg/dL or .mu.mon) or urine output
(mL/d)).
[0423] The baseline SOFA score can be assumed to be zero in
patients not known to have preexisting (acute or chronic) organ
dysfunction before the onset of infection. A SOFA score of at least
2 points reflects an overall mortality risk of approximately 10% in
a general hospital population with suspected infection (Singer et
al., JAMA. 2016 Feb. 23; 315(8):801-10).
[0424] In one embodiment, the human subject is suffering from SIRS,
sepsis or septic shock associated with a SOFA score of at least 2
points.
[0425] In one embodiment, the human subject is suffering from SIRS,
sepsis or septic shock associated with an organ dysfunction defined
as an acute change in his/her SOFA score of at least 2 points.
[0426] The presence of organ dysfunction associated with sepsis or
septic shock may also be assessed using the quick SOFA score (also
referred to as quickSOFA or qSOFA). The qSOFA scoring system relies
on three criteria: respiratory rate .gtoreq.22 breaths/min; altered
mentation (Glasgow coma scale <15); and systolic blood pressure
.gtoreq.100 mm Hg (Seymour et al., JAMA. 2016 Feb. 23;
315(8):762-74).
[0427] In one embodiment, the human subject is suffering from SIRS,
sepsis or septic shock associated with a qSOFA score of at least 2
points.
[0428] In one embodiment, the human subject is suffering from SIRS,
sepsis or septic shock associated with an organ dysfunction defined
as an acute change in his/her qSOFA score of at least 2 points.
[0429] According to one embodiment, the human subject is suffering
from septic shock.
[0430] According to one embodiment, the human subject is suffering
from SIRS, sepsis or septic shock, preferably septic shock, and
receiving the standard of care. The standard of care, in particular
the standard of care for septic shock, may include, without being
limited to, fluid therapy, vasopressor therapy as described
hereinabove, cardiovascular support, respiratory support (such as
mechanical ventilation), renal support and/or sedation. Thus, in
one embodiment, the human subject is suffering from suffering from
SIRS, sepsis or septic shock, preferably septic shock, and
receiving fluid therapy, vasopressor therapy as described
hereinabove, cardiovascular support, respiratory support (such as
mechanical ventilation), renal support and/or sedation.
[0431] According to the present invention, the human subjects
suffering from an inflammatory disorder, preferably SIRS, sepsis or
septic shock identified with the method as described hereinabove
are susceptible to respond to an administration of a therapy, in
particular of a TREM-1 inhibitor; as described hereinabove.
[0432] According to one embodiment, the aim of the method described
hereinabove is to identify human subjects suffering from an
inflammatory disorder, preferably SIRS, sepsis or septic shock who
are "responder", i.e., who respond or are susceptible to respond to
an administration of a therapy, in particular of a TREM-1
inhibitor, as described hereinabove.
[0433] Conversely, in the present invention, human subjects
suffering from an inflammatory disorder, preferably SIRS, sepsis or
septic shock, who are "non-responder" are human subjects who do not
respond or are not susceptible to respond to an administration of a
therapy, in particular of a TREM-1 inhibitor, as described
hereinabove.
[0434] According to one embodiment, for a human subject suffering
from an inflammatory disorder, preferably SIRS, sepsis or septic
shock, a response to the administration of a therapy, in particular
of a TREM-1 inhibitor, as described hereinabove is characterized by
at least one of the following occurring after the administration of
the TREM-1 inhibitor: [0435] a reversal of a hypotensive shock,
preferably within or after the administration period of the
therapy, preferably a TREM-1 inhibitor, for example over the 6, 12,
18 or 24 hours following the end of said administration, wherein a
shock reversal is defined as the absence of any vasopressor therapy
during 24 hours (i.e., not requiring to restart a vasopressor
therapy in the 24 hours following the end of a vasopressor
therapy); [0436] a decrease of a severity score used to assess the
severity of the disease and/or the prognosis of a human subject
suffering from an inflammatory disorder, preferably SIRS, sepsis or
septic shock upon admission in ICU or emergency unit, such as the
APACHE II score, the APACHE III score, the APACHE IV score, the
SAPS score, the SAPS II score or the SAPS 3 score; [0437] a
decrease of an organ dysfunction score used to assess the presence
of organ dysfunction in a human subject suffering from an
inflammatory disorder, preferably SIRS, sepsis or septic shock upon
admission in ICU or emergency unit, such as the SOFA score, the
qSOFA score, the MODS (Multiple Organ Dysfunction Score), the
P-MODS (Pediatric Multiple Organ Dysfunction Score) or the LODS
(Logistic Organ Dysfunction System); [0438] a decrease of the SOFA
score and/or of the qSOFA score, in particular over the 1, 2, 3, 4,
5, 6 or 7 day(s) following the start of the administration of the
therapy, preferably a TREM-1 inhibitor, preferably with reference
to the SOFA score and/or the qSOFA score assessed upon admission in
ICU or emergency unit or before the start of the administration of
the therapy; in one embodiment, said decrease of the SOFA score is
a decrease of at least 1 point (also referred as a delta of -1
point or .DELTA.SOFA of -1 point), preferably of at least 1.5
point, in particular at day 3 or at day 5 following the start of
the administration of the therapy, preferably with reference to the
SOFA score and/or the qSOFA score assessed upon admission in ICU or
emergency unit or before the start of the administration of the
therapy; [0439] a decrease in the requirement for cardiovascular
support, for example a decrease in the use of vasopressor therapy,
in particular over the 1, 2, 3, 4, 5, 6 or 7 day(s) following the
start of the administration of the therapy, preferably a TREM-1
inhibitor, preferably with reference to the requirement for
cardiovascular support upon admission in ICU or emergency unit or
before the start of the administration of the therapy; [0440] a
decrease in the requirement for respiratory support, for example a
decrease in the use of invasive mechanical ventilation (IMV), in
particular over the 1, 2, 3, 4, 5, 6 or 7 day(s) following the
start of the administration of the therapy, preferably a TREM-1
inhibitor, preferably with reference to the requirement for
respiratory support upon admission in ICU or emergency unit or
before the start of the administration of the therapy; [0441] a
decrease in the requirement for renal support, for example a
decrease in the use of continuous or discontinuous renal
replacement therapy also referred to as RRT (e.g., dialysis), in
particular over the 1, 2, 3, 4, 5, 6 or 7 day(s) following the
start of the administration of the therapy, preferably a TREM-1
inhibitor, preferably with reference to the requirement for renal
support upon admission in ICU or emergency unit or before the start
of the administration of the therapy; [0442] a decrease in the risk
of reinfection, in particular a decrease in the risk of reinfection
in the 28, 90 or 365 days following the initial inflammatory
disorder, in particular the initial infection responsible for an
inflammatory disorder; [0443] an absence of reinfection, in
particular an absence of reinfection in the 28, 90 or 365 days
following the initial inflammatory disorder, in particular the
initial infection responsible for an inflammatory disorder; [0444]
a decrease in the risk of rehospitalization, in particular a
decrease in the risk of reinfection in the 28, 90 or 365 days
following the initial inflammatory disorder, in particular the
initial infection responsible for an inflammatory disorder; [0445]
an absence of rehospitalization, in particular an absence of
rehospitalization in the 28, 90 or 365 days following the first
hospitalization; [0446] an increase in the chance of survival, in
particular 1-year, 2-year, 3-year, 4-year, 5-year, 6-year, 7-year,
8-year, 9-year or 10-year survival, following the start of the
administration of the therapy, preferably a TREM-1 inhibitor; in
one embodiment the chance of survival, in particular in a subject
with multiple comorbidities, is assessed with the Charlson
Comorbidity Index (CCI) and an increase in the chance of 10-year
survival corresponds to a decrease of the Charlson Comorbidity
Index, preferably with reference to CCI assessed upon admission in
ICU or emergency unit or before the start of the administration of
the therapy; [0447] a decrease in the risk of sepsis-related death,
in particular at day 5, day 28, day 90 or day 365 following the
start of the administration of the therapy, preferably a TREM-1
inhibitor; [0448] a decrease in the risk of the all-cause death, in
particular at day 5, day 28, day 90 or day 365 following the start
of the administration of the therapy, preferably a TREM-1
inhibitor; [0449] a decrease in the risk of post-sepsis or
post-shock morbidity, in particular at day 5, day 28, day 90 or day
365 following the start of the administration of the therapy,
preferably a TREM-1 inhibitor; [0450] an increase in the quality of
life, in particular in the post-sepsis or post-shock quality of
life that may be assessed for example through an evaluation of
survival and quality-adjusted life years (QALYs), estimated from
the EQ5D. For example, health-related quality of life (HRQoL)
scores from the EQ 5D 5L may be calculated and converted to utility
scores, in particular at 3 months, 6 months, 9 months, 12 months,
18 months, 24 months or 36 months following the start of the
administration of the therapy; [0451] a decrease in the level of an
inflammatory marker, such as, for example, CRP or IL6, IL-8, IL-10,
MCP-1 and TNF-.alpha.), in particular over the 1, 2, 3, 4, 5, 6 or
7 day(s) following the start of the administration of the therapy,
preferably a TREM-1 inhibitor, preferably with reference to the
level assessed upon admission in ICU or emergency unit or before
the start of the administration of the therapy; or [0452] a
decrease in the level of an endothelial injury marker, such as, for
example, Ang-2, VCAM-1, VGEFR-1 and E-selectin, in particular over
the 1, 2, 3, 4, 5, 6 or 7 day(s) following the start of the
administration of the therapy, preferably a TREM-1 inhibitor,
preferably with reference to the level assessed upon admission in
ICU or emergency unit or before the start of the administration of
the therapy.
[0453] In one embodiment, a human subject suffering from SIRS,
sepsis or septic shock, preferably septic shock, susceptible to
respond to the administration of a therapy, in particular of a
TREM-1 inhibitor, as described hereinabove, i.e., a responder, is a
human subject suffering from SIRS, sepsis or septic shock,
preferably septic shock, susceptible to reverse a hypotensive
shock, preferably within or after the administration period of the
therapy, in particular of a TREM-1 inhibitor, for example over the
6, 12, 18 or 24 hours following the end of said administration,
wherein a hypotensive shock reversal is defined as the absence of
any vasopressor therapy during 24 hours (i.e., not requiring to
restart a vasopressor therapy in the 24 hours following the end of
a vasopressor therapy).
[0454] Conversely, in one embodiment, a human subject suffering
from SIRS, sepsis or septic shock, preferably septic shock, not
susceptible to respond to the administration of a therapy, in
particular of a TREM-1 inhibitor, as described hereinabove, i.e., a
non-responder, is a human subject suffering from SIRS, sepsis or
septic shock preferably septic shock, unable to reverse a
hypotensive shock, preferably within or after the administration
period of the therapy, in particular of a TREM-1 inhibitor, for
example over the 6, 12, 18 or 24 hours following the end of said
administration, wherein a hypotensive shock reversal is defined as
the absence of any vasopressor therapy during 24 hours, or is a
human subject suffering from SIRS, sepsis or septic shock,
preferably septic shock, requiring the restarting of a vasopressor
therapy within the 6, 12, 18 or 24 hours following withdrawal of
the therapy, in particular of a TREM-1 inhibitor.
[0455] In one embodiment, a responder is a human subject suffering
from SIRS, sepsis or septic shock, who following the administration
of a TREM-1 inhibitor as described hereinabove, is susceptible to
have his/her sequential organ failure assessment (SOFA) score
decrease.
[0456] In one embodiment, a responder is a human subject suffering
from SIRS, sepsis or septic shock, who following the administration
of a TREM-1 inhibitor as described hereinabove, is susceptible to
have his/her sequential organ failure assessment (SOFA) score
decrease with reference to the SOFA score assessed upon admission
in ICU or emergency unit or before the start of the administration
of the therapy.
[0457] As used herein, the difference in the SOFA score following
the administration of a TREM-1 inhibitor, in particular with
reference to the SOFA score assessed upon admission in ICU or
emergency unit or before the start of the administration of the
therapy, may also be referred to as the delta SOFA score or
.DELTA.SOFA score.
[0458] In one embodiment, a responder is a human subject suffering
from SIRS, sepsis or septic shock, who following the administration
of a TREM-1 inhibitor as described hereinabove, is susceptible to
have his/her SOFA score decrease by at least 1 point, preferably by
at least 1.5 point, in particular with reference to the SOFA score
assessed upon admission in ICU or emergency unit or before the
start of the administration of the therapy.
[0459] Thus, in one embodiment, a responder is a human subject
suffering from SIRS, sepsis or septic shock, who following the
administration of a TREM-1 inhibitor as described hereinabove, is
susceptible to have a delta SOFA score (.DELTA.SOFA score) of at
least -1 point (minus 1 point), preferably at least -1.5 point
(minus 1.5 point).
[0460] In one embodiment, said decrease of the SOFA score, in
particular the decrease by at least 1 point, preferably at least
1.5 point, is the decrease assessed at day 1, 2, 3, 4, 5, 6 or 7
following the start of the administration of the therapy,
preferably at day 3 or at day 5 following the start of the
administration of the therapy.
[0461] Thus, in one embodiment, said delta SOFA score (.DELTA.SOFA
score) is the delta SOFA score assessed at day 1, 2, 3, 4, 5, 6 or
7 following the start of the administration of the therapy,
preferably at day 3 or at day 5 following the start of the
administration of the therapy.
[0462] With the present application, the Applicant shows that
measuring the level of soluble TREM-1 (sTREM-1) in a biological
sample from human subjects suffering from septic shock and
comparing their measured level of sTREM-1 to a predetermined
sTREM-1 value allows to identify the human subjects suffering from
septic shock susceptible to respond to a TREM-1 inhibitor. In
particular, the Applicant shows that human subjects suffering from
septic shock with a circulatory level of sTREM-1 higher than the
median sTREM-1 level predetermined in a reference population of
human subjects suffering from septic shock are more likely to
respond to, and thus benefit from, the administration of a TLT-1
peptide inhibiting TREM-1 activity.
[0463] A prompt patient care is critical for the prognosis of
subjects suffering from SIRS, sepsis or septic shock.
[0464] Measuring the level of sTREM-1, in particular the level of
sTREM-1 protein, can be carried out very rapidly, notably with a
point-of-care testing (POCT) or bedside testing, or with a central
laboratory assay (near-to-patient assay). The level of sTREM-1 may
thus be measured shortly following the onset of SIRS, sepsis or
septic shock; following the diagnosis of the human subject with
SIRS, sepsis or septic shock; following the hospitalization, in
particular the admission in ICU or emergency unit, of the human
subject for SIRS, sepsis or septic shock; or following the start of
vasopressor therapy.
[0465] Results may be thus available within the first 2, 3, 6, 9,
12, 15, 18, 21 or 24 hours, and a care plan may be decided upon
immediately thereafter.
EXAMPLES
[0466] The present invention is further illustrated by the
following examples.
Example 1
[0467] Materials and Methods
[0468] Cell Culture and Stimulation
[0469] The human myelomonocytic cell lines U937 (Culture
Collections, Public Health England No. 85011440) were cultured in
RPMI 1640 Glutamax supplemented with 10% fetal calf serum, 25 mM
Hepes, and 100 U/ml penicillin and streptomycin (all from Thermo
Fisher Scientific, USA). For some experiments, U937 cells were
cultured under the same conditions supplemented with 100 nM of
1,25-dihydroxyvitamin D3 (Sigma-Aldrich, USA) to induce
upregulation of TREM-1 (U937-vitD cells). For shedding experiments,
Pro-MMP-9 was activated by pre-incubation with 1 mM APMA for 24 h
at 37.degree. C. Cells were seeded at 0.5.times.10.sup.6 cells/mL
and stimulated with LPS from Escherichia coli 0127:B8 (10 .mu.g/mL;
Sigma-Aldrich), Pro-MMP-9 (1 .mu.g/mL; R&D systems, Abingdon,
UK), p-aminophenylmercuric acetate (APMA; 1 mM; Sigma-Aldrich), and
APMA-activated MMP-9 (1 .mu.g/mL) at 37.degree. C. for 30 min. Cell
supernatants were collected and sTREM-1 was measured using Human
TREM-1 Quantikine ELISA Kit (Biotechne, R&D Systems) according
to the manufacturer's instructions.
[0470] FACS Analysis
[0471] TREM-1 expression was detected by FACS after staining with 5
.mu.L allophycocyanin (APC) conjugated mouse monoclonal anti-human
TREM-1 or the corresponding isotype-APC antibodies (Miltenyi
Biotec, Germany) for 30 min at 4.degree. C. in dark. After washing
with PBS twice, cells were re-suspended and fixed with 4%
paraformaldehyde. Accuri C6 flow cytometer (Becton Dickinson, San
Jose, Calif., USA) was used for analysis.
[0472] Confocal Microscopy
[0473] Cells were seeded at 0.3.times.10.sup.6 cells/well and
stimulated with LPS from Escherichia coli 0127:B8 (1 .mu.g/mL;
Sigma-Aldrich) on LabTek chambers (Thermo Fisher Scientific) for 1
h. After stimulation, cells were washed and fixed with
paraformaldehyde (4%) for 20 min and permeabilized with Triton 0.1%
for 30 min prior to incubation with the primary antibodies
(anti-hTREM-1-AF488; Bioss, USA) at 4.degree. C. overnight. Nuclei
were stained with TO-PRO-3 (1 .mu.g/mL; Invitrogen, USA) for 1 h at
37.degree. C. After washing, coverslips were mounted using a
solution of Vectashield (Vector Laboratories, USA). Confocal images
were obtained using a Leica SP5 confocal laserscanning microscope
system (Leica, Germany) fitted with appropriated filter sets and
acquired in sequential scan mode.
[0474] Results
[0475] sTREM-1 is a Marker of TREM-1 Activation.
[0476] We had previously described that TREM-1 dimerization is
essential for its activation on innate immune cells (Carrasco et
al., Cell Mol Immunol. 2018 Mar. 22). It is triggered by the
initial engagement of TLRs. Indeed, stimulation of human primary
monocytes and neutrophils was able to induce the mobilization and
clustering of TREM-1 at the membrane. The dimeric configuration of
TREM-1 was thus identified to be the active conformation of the
receptor, able to bind its endogenous ligand and able to induce
activation of downstream intracellular pathway. It has been
previously demonstrated that metalloproteinases are responsible for
shedding of the TREM-1 ectodomain through proteolytic cleavage of
its long juxtamembrane linker (Gomez-Pina et al., J Immunol. 2007
Sep. 15; 179(6):4065-73). We demonstrated here that this shedding
necessitates TREM-1 dimerization, suggesting that the release of
sTREM-1 is only possible when the receptor is in its active
conformation. TREM-1 is expressed at very low levels on resting
U937 cells, and it is upregulated when vitamin D is added to the
culture media (FIG. 1A). To investigate whether the proteolytic
cleavage of TREM-1 and subsequent release of sTREM-1 was linked to
the active conformation of TREM-1, we measured sTREM-1 in the
supernatant of U937 and U937-vitD cells, after 15-min incubation
with LPS and/or Act-MMP9. TREM-1 expression was diffuse at the
membrane of resting U937-vitD cells, and LPS was able to induce the
clustering and dimerization of TREM-1 at the membrane (FIG. 1B,
white arrows). No sTREM-1 was detectable in the supernatant of U937
cells, neither after LPS stimulation nor addition of activated-MMP9
(Act-MMP9). Incubation of U937-vitD cells with LPS or Act-MMP9
alone was not associated with a release of sTREM-1. Interestingly,
sTREM-1 was released by U937-vitD cells only in the condition where
U937-vitD were co-stimulated with LPS and Act-MMP9 ( FIG. 1C).
These results confirm that the MMP9-induced proteolytic cleavage of
TREM-1 is only possible when TREM-1 is in its dimeric state, which
is the active conformation of the receptor. sTREM-1 may thus be a
reliable biomarker of TREM-1 receptor activation.
Example 2
[0477] Materials and Methods
[0478] sTREM-1 Measurements: Validated ELISA Method for the
Quantitative Determination of sTREM-1 in Human K2-EDTA Plasma
[0479] The quantitative determination of sTREM-1 was carried out
using a commercial kit (Human TREM-1 Quantikine ELISA, BioTechne,
R&D Systems) and quality control (QC) samples at 3
concentration levels: QC Low (101 pg/mL), QC Mid (532 pg/mL) and QC
High (1080 pg/mL) for routine assay.
[0480] A pre-study assay qualification was performed to determine
the intra-run precision and accuracy of the assay. A pre-study
assay qualification run consists in calibration standards and QC
samples at 5 concentration levels (LLOQ (41.3 pg/mL), Low, Mid,
High and ULOQ (1440 pg/mL), each QC samples must be processed in
sextuplate per assay. For each concentration level, the final
calculation of intra-run imprecision and inaccuracy must not use
less than 5 out of the 6 values. The intra-run imprecision must be
less than or equal to 20% (25% at the LLOQ and ULOQ) and the
intra-run inaccuracy must be within .+-.20% (25% at the LLOQ and
ULOQ). Pre-study assays must be validated 3 times before starting
routine assays.
[0481] Standard (STD) samples were prepared freshly each analysis
day using Calibrator diluent RD5-18 and the Human TREM-1 standard
from the kit. STD samples were prepared to obtain 10 points in the
calibration curve: 3000, 2000, 1000, 750, 500, 250, 125, 62.5, 31.3
and 15.6 pg/mL. Calibration standards at 3000 and 15.6 pg/mL were
used as anchoring points (Lowest calibration standard (LCS) and
Higher calibration standard (HCS) of the calibration curve.
[0482] 100 .mu.L of Assay Diluent RD1-27 were added to each well.
50 .mu.L of Standard, QC or Samples were then added per well. Plate
are incubated for 2 hours at room temperature on a microplate
shaker at 450 rpm. After that 2-hour incubation, plates were
manually washed 3 times with Wash Buffer, and 200 .mu.L of Human
TREM-1 Conjugate were added to each well. Plate were incubated for
2 supplementary hours at room temperature on a microplate shaker at
450 rpm. After washing, 200 .mu.L of Substrate Solution were added
to each well. Plate were incubated for 30 minutes at room
temperature in the dark. Optical Density (OD) was read at 650 nm
(pre-read), and when the OD(650 nm) of HCS was about 1.0, 50 .mu.L
of Stop Solution were added to each well. Then, the OD(450 nm) of
each well within 30 minutes was measured.
[0483] The calibration curve was obtained by plotting the optical
density versus the concentrations of the calibration standards,
using the logistic curve (4-PL) and 1/y2 weighting factor.
[0484] Calibration standards from 2000 to 31.3pg/mL were excluded
from the final calibration curve only if the back-calculated
inaccuracy was outside .+-.20% (.+-.25% at the LCS and HCS), or for
reasons of instrument failure, or for a documented problem during
preparation or assay, or if the coefficient of variation (CV %) on
concentration on replicate determination (n=2 wells per calibration
standard) was higher than 20%. No more than 30% of these
calibration standards may be excluded from each series of
calibration standards and the final calibration curve must contain
at least 6 non-zero calibration concentration levels including the
LCS and the HCS. Blank (0.00 pg/mL), which consisted in Calibrator
Diluent RD5-18, was not used for the calculation of the calibration
curve.
[0485] Each routine run included 6 QC samples (n=2 in duplicate at
3 concentration levels). At least 4 of the 6 QC samples must be
within acceptance criteria (with at least one value within
acceptance criteria for each concentration level). Acceptance
criteria were defined as follows: the individual inaccuracy
calculated on the concentration from duplicate determination (n=2
wells per sample) must be within .+-.20% compared to the nominal
value. The coefficient of variation (CV %) on the concentration of
replicate determination must be less than or equal to 20%.
[0486] The coefficient of variation (CV %) on the concentration of
replicate determination (n=2 wells per test sample) of test samples
must also be less than or equal to 20%.
[0487] Evaluation of sTREM-1 in the AdrenOSS Cohort
(NCT02393781)
[0488] This study was an ancillary investigation of septic shock
patients from the AdrenOSS study (NCT02393781), a European
prospective, observational, multinational study in 24 centers from
5 countries (France, Belgium, the Netherlands, Italy, and Germany).
Patients were recruited from June 2015 to May 2016. The study
protocol was approved by the local ethical committees and conducted
in accordance with Directive 2001/20/EC, as well as Good Clinical
Practices (I.C.H. version 4 of May 1, 1996 and Decision of Nov. 24,
2006) and the Declaration of Helsinki. The patients who were
enrolled were 18 years and older and were (i) admitted to the ICU
for sepsis or septic shock or (ii) transferred from another ICU in
the state of sepsis and septic shock within less than 24 hours
after primary admission. If patients were treated with
vasopressors, they were considered eligible only if treatment had
been started within a maximum of 24 hours after the primary
admission before ICU admission. When patients were included, they
were stratified by severe sepsis and septic shock based on the
definitions for sepsis and organ failure from 2001. Exclusion
criteria were pregnancy, vegetative coma, and participation in an
interventional trial in the preceding month. Informed consent was
obtained from all patients or their lawful representatives prior to
enrollment in the study. Patients were treated according to current
guidelines, and all treatments and procedures were registered.
sTREM-1 was measured in blood samples withdrawn at baseline (also
referred to as day 1 or d1), day 2 (d2) and day 3 (d3).
[0489] Statistical Analysis for the AdrenOSS Cohort
[0490] Analysis was performed on 293 septic shock patients. The
demographic and baseline characteristics are summarized in a
descriptive fashion within each population: categorical variables
were summarized by contingency tables (frequencies and
percentages). Continuous variables were summarized by their mean,
standard deviation, standard error, median, quartiles and range,
minimum and maximum values. sTREM-1 levels at day 1 (independently
referring to baseline, or entry to ICU) were also described (mean,
standard deviation, standard error, median, quartiles and range,
minimum and maximum values) and separately within the population
and subgroups as defined. 28-day mortality rates were estimated
within each quartile of the distribution of sTREM-1 levels at day
1. Time to death was analyzed in a proportional hazards COX
regression model. A ROC Area Under Curve (AUC) with its 95% CI to
predict 28-day mortality from sTREM-1 day 1 levels was calculated.
An optimal sTREM-1 cut-off was estimated based i) on the
minimization of the distance between the point (0,1) on the upper
left-hand corner of ROC space and any point on ROC curve and ii) on
the Youden index. Boxplots of sTREM-1 by survivor/non-survivors and
RRT (renal replacement therapy) were plotted. When necessary,
sTREM-1 data were log transformed. The association of sTREM-1
levels at day 1 and 28-day mortality was assessed in univariable
and multivariable logistic regression models adjusted for sTREM-1
at baseline (day 1) as well as adjusted for other prognostic
factors (age, gender, cardiac comorbidities, non-cardiac
comorbidities and lactate (maximum or minimum on day 1), SOFA at
D1, APACHE II at D1, SAPS II at D1.
[0491] Results
[0492] sTREM-1 is a Prognostic Marker in Septic Shock Patients:
Results from the AdrenOSS Cohort
[0493] The demographics and general baseline characteristics of
patients in the whole population, and in each sTREM-1 quartiles are
depicted in Table 1 below. No significant differences were observed
in patients between the four sTREM-1 quartile groups (Q1, Q2, Q3
and Q4) for baseline characteristics, age, gender, BMI (Body Mass
Index), type of ICU admission, origin of sepsis, medical history,
and physiological values at admission. The first quartile group,
i.e., Q1, comprises patients who had a sTREM-1 level lower than the
first quartile value (299 pg/mL), that is to say patients who had a
sTREM-1 level between 42 and 299 pg/mL. The second quartile group,
i.e., Q2, comprises patients who had a sTREM-1 level between the
first quartile value (299 pg/mL) and the second quartile value (497
pg/mL), that is to say patients who had a sTREM-1 level between 299
pg/mL and 497 pg/mL. The third quartile group, i.e., Q3, comprises
patients who had a sTREM-1 level between the second quartile value
(497 pg/mL) and the third quartile value (809 pg/mL), that is to
say patients who had a sTREM-1 level between 497 pg/mL and 809
pg/mL. The fourth quartile group, i.e., Q4, comprises patients who
had a sTREM-1 level higher than the third quartile value (809
pg/mL), that is to say patients who had a sTREM-1 level between 809
pg/mL and 5540 pg/mL. There was a significant correlation between
sTREM-1 values and lactate levels, arterial pH, creatinine levels,
urea levels and PCT levels at baseline. sTREM-1 levels were also
correlated with renal replacement therapy (RRT), SOFA, SAPS II and
APACHE II scores at baseline, as well as with 28-day and 90-day
mortality. 28-day mortality rates were 12.2%, 30.1%, 37% and 49.3%
in quartiles Q1, Q2, Q3 and Q4, respectively. Overall mortality
rate was 32%. These results confirm that the level of TREM-1
pathway activation at entry to ICU, assessed through the
measurement of plasma sTREM-1, is associated with a more
complicated outcome in septic shock patients.
TABLE-US-00001 TABLE 1 Demographic and General Characteristics,
comorbidities, chronic medications, clinical signs at admission of
patients from the AdrenOSS cohort. SOFA.4 = SOFA score without
liver and CNS component sTREM-1 Q1 sTREM-1 Q2 sTREM-1 Q3 sTREM-1 Q4
all (42, 299] (299, 497] (497, 809] (809, 5540] Characteristics n =
293 n = 74 n = 73 n = 73 n = 73 p-value sTREM-1 at 497 [299-809]
admission (pg/ml) Age (year) 67.2 [57.8-77.1] 63.2 [52.5-70.0] 69.8
[60.0-76.4] 69.9 [58.7-81.1] 66.7 [60.4-77.1] 0.0055 Females (No.
%) 106 (36.2) 22 (29.7) 27 (37) 28 (38.4) 29 (39.7) 0.5931 Body
Mass Index 25.9 [23.0-30.4] 25.9 [22.9-29.6] 25.4 [22.5-29.0] 26.7
[23.3-30.0] 28.4 [23.9-33.3] 0.1056 (kg/m2) Type of ICU 0.1452
admission: Medical 215 (73.4) 55 (74.3) 53 (72.6) 54 (74) 53 (72.6)
Surgical - emergency 67 (22.9) 19 (25.7) 14 (19.2) 15 (20.5) 19
(26) procedure Surgical - elective 11 (3.8) 0 (0) 6 (8.2) 4 (5.5) 1
(1.4) procedure Orgin of sepsis: 0.4141 Lung 81 (27.6) 26 (35.1) 24
(32.9) 16 (21.9) 15 (20.5) Blood stream 40 (13.7) 5 (6.8) 8 (11) 14
(19.2) 13 (17.8) Urinary tract 50 (17.1) 9 (12.2) 13 (17.8) 14
(19.2) 14 (19.2) Catheter 19 (6.5) 4 (5.4) 4 (5.5) 8 (11) 3 (4.1)
Peritonitis 17 (5.8) 4 (5.4) 3 (4.1) 6 (8.2) 4 (5.5) Endocarditis
18 (6.1) 6 (8.1) 4 (5.5) 3 (4.1) 5 (6.8) Bile duct infection 4
(1.4) 1 (1.4) 1 (1.4) 2 (2.7) 0 (0) CNS 3 (1) 2 (2.7) 0 (0) 0 (0) 1
(1.4) Other 61 (20.8) 17 (23) 16 (21.9) 10 (13.7) 18 (24.7) Medical
history OR specific diseases (COPD, DMII, immunodysfunction,
chronic renal failure, ischemic or congestive heart disease) any
cardiac 213 (72.7) 47 (63.5) 49 (67.1) 59 (80.8) 58 (79.5) 0.0386
comorbidity (yes) any non-cardiac 226 (77.1) 56 (75.7) 59 (80.8) 56
(76.7) 55 (75.3) 0.8513 comorbidity (yes) any chronic 204 (69.6) 51
(68.9) 53 (72.6) 52 (71.2) 48 (65.8) 0.8182 medication (yes)
Physiological values at admission Temperature (.degree. C.) 37.1
[36.1-38] 37.4 [36.6-38.3] 37.15 [36.27-37.92] 37.1 [36.3-37.9]
36.6 [35.8-37.4] 0.0052 Mean blood 103 106.5 106 [92.5-120] 102
[85-118] 100 [83-125.5] 0.3179 pressure (mmHg) [89.25-124.75]
[92-135.75] Heart rate (bpm) 56 [46-68] 61.5 [48-71.75] 56 [48-68]
54 [45-68] 53 [43.75-65] 0.2095 Central Venous 71 [60-84] 72.5
[65-93] 73 [64-82] 71.5 [56-79.75] 66 [58-82.5] 0.0636 pressure
(mmHg) Glasgow score 21 [18-26.5] 21 [18-27] 22 [17.75-27] 21
[18-26] 22 [18-26] 0.9260 Fluid Balance (ml) 105 [92-121.25] 109.5
[95-124.5] 99.5 [92-116.25] 105 [86-124] 105 [86-121] 0.5021
PaO2/FiO2 205 [128-326] 217 [123-354] 225.5 179.5 228.5 0.5418
[134.5-287.75] [132.25-294.75] [121.5-330.5] Laboratory values at
admission Lactate (mmol/L) 3.0 [2.0-5.2] 2.5 [1.5-3.4] 2.7
[2.0-4.7] 3.5 [2.0-5.7] 4.2 [2.7-9.7] <0.0001 Aterial pH 7.35
[7.27-7.42] 7.4 [7.33-7.46] 7.36 [7.29-7.43] 7.32 [7.26-7.4] 7.29
[7.18-7.38] <0.0001 Bilirubin (.mu.mol/L) 11 [6-21] 10.5
[7-18.25] 11 [7-22.5] 10 [6-19] 12 [6-21.5] 0.9628 Platelets
(109/L) 190 [114-269] 186 [108.25-251] 210 [122-287] 191 [123-239]
159 [107-291] 0.6676 Creatinine (mg/dL) 1.65 [1.06-2.72] 0.93
[0.68-1.3] 1.4 [1.06-1.96] 2.08 2.66 [1.79-4.21] <0.0001
[1.53-3.27] BUN or Urea 72 [46.65-120] 52.25 64.26 92 111.11
[60.66-149] <0.0001 (mg/dL) [33.03-67.2] [44.44-102.1]
[60.06-130.93] Hematocrit (%) 34 [29-38] 32 [28-36] 32.5 [28.75-37]
35 [29-39] 34 [29-39] 0.1827 White blood count 12550 [6270-18700]
10940 [6467-16827] 12000 [7135-17000] 11945 [5135-18287] 14600
[8300-22120] 0.2185 (per mm3) Troponin T, 55 [25-232.5] 37.86 33.5
[20.75-160.5] 54 [39-200.5] 133.1 [68-245.5] 0.1605 maximum on day
1 [17.25-160] Troponin I, 130 [48.5-395.5] 175 63.1 232.95 84.7
0.1402 maximum on day 1 [52.5-381.57] [31.33-192.45]
[113.72-612.02] [39.28-583.25] PCT, maximum on 17.66 8.03 7.51
[3.92-24] 31 [8.42-84] 57 [17.62-113.5] <0.0001 day 1 (ng/mL)
[4.49-66.61] [2.59-25.88] BNP, maximum 383 [112-1262] 210.3 179.8
[87-257] 1639 376.2 [138.5-881] 0.0091 on day 1 [102.25-580.25]
[1260.05-2208.85] NT-proBNP, 7957 4162 [519-10203] 5428
[2783-13924] 9757 11108 [7209-22440] 0.0399 maximum on day 1
[2764-16926] [4170-19402] Organ support at admission Mechanical
0.1110 ventilation: invasive 149 (50.9) 34 (45.9) 34 (46.6) 36
(49.3) 45 (61.6) non-invasive 47 (16) 8 (10.8) 12 (16.4) 14 (19.2)
13 (17.8) none 97 (33.1) 32 (43.2) 27 (37) 23 (31.5) 15 (20.5)
Renal replacement 38 (13) 1 (1.4) 5 (6.8) 9 (12.3) 23 (31.5)
<0.0001 therapy (yes) Organ dysfunction scores SOFA (points) 9
[7-11] 7 [6-10] 9 [7-11] 10 [8.5-12] 11 [9-13] <0.0001 SOFA.4
(points) 8 [7-10] 7 [6-8.75] 8 [6-9] 9 [7-10] 10 [8-11] <0.0001
SAPS2 (points) 58 [46-71] 42 [42.25-60] 55 [44-69] 59 [48-71] 65
[57-77] <0.0001 APACHE 2 (points) 18 [13-22] 15 [11-18.75] 15
[12-20] 18 [15-23] 21 [17-24] <0.0001 Mortality (%) 28-day,
deaths 94 (31.1) 9 (12.2) 22 (30.1) 27 (37) 36 (49.3) <0.0001
90-day, deaths 116 (39.6) 14 (18.9) 24 (32.9) 36 (49.3) 42 (57.5)
<0.0001 * p-value from non-parametric Kruskal-Wallis or
Chi.sup.2 test, respectively.
[0494] FIG. 2A depicts the ROC plot. FIG. 2B describes the survival
of the patients over time in form of a Kaplan-Meier Curve. Patients
with the lowest sTREM-1 values (Q1 quartile group) had a better
survival at 28-day than patients with the highest sTREM-1 values
(Q2 to Q4 quartile groups). The survival rates within the sTREM-1
quartile groups were 87.8%, 69.9%, 63%, and 50.7% from Q1 to Q4,
respectively. An optimal cutoff value was calculated at 408 pg/mL
for predicting mortality from the data presented in Table 2
below.
TABLE-US-00002 TABLE 2 Cut off shifting: sensitivity and
specificity for last timepoint (28 days) based on Nearest Neighbor
Estimation (NNE) and Kaplan-Meier (KM) method. Cutoff Quantile
NNE.Sens NNE.Spec sumNNE KM.Sens KM.Spec sumKM -Inf 0 100 0 100 100
0 100 121 1.7 99.1 2 101.1 98.9 2 100.9 155 3.4 98.3 4.1 102.4 98.9
4.5 103.4 178 5.5 97.5 6.7 104.2 96.8 6.5 103.3 186 7.5 96.6 9.2
105.8 95.7 9 104.7 201 9.6 95.5 11.6 107.1 95.7 12.1 107.8 210 11.3
94.6 13.6 108.2 95.7 14.6 110.3 231 13.7 93.8 16.6 110.4 94.7 17.6
112.3 241 16.4 92.8 20.1 112.9 92.6 20.6 113.2 254 19.1 91.8 23.5
115.3 91.5 24.1 115.6 268 20.8 91.1 25.6 116.7 91.5 26.6 118.1 282
22.5 90.3 27.7 118 91.5 29.1 120.6 296 24.9 89.5 30.7 120.2 90.4
32.2 122.6 307 27 88.7 33.2 121.9 89.4 34.7 124.1 323 28.7 87.9
35.3 123.2 89.4 37.2 126.6 330 30.7 86.7 37.7 124.4 87.2 39.2 126.4
340 33.4 85.1 40.9 126 87.2 43.2 130.4 349 35.5 83.6 43.2 126.8
85.1 45.2 130.3 372 38.2 81.3 46.1 127.4 84 48.7 132.7 380 40.3
79.4 48.2 127.6 81.9 50.8 132.7 396 42.3 77.5 50.3 127.8 79.8 52.8
132.6 409 44 75.7 51.9 127.6 76.6 53.8 130.4 444 46.1 73.1 53.8
126.9 72.3 54.8 127.1 467 48.1 70.4 55.5 125.9 71.3 57.3 128.6 490
49.8 68 57 125 68.1 58.3 126.4 515 51.9 65.3 58.8 124.1 64.9 59.8
124.7 536 53.6 63.2 60.3 123.5 63.8 61.8 125.6 553 56 60.3 62.5
122.8 61.7 64.3 126 577 58 57.9 64.4 122.3 60.6 66.8 127.4 616 60.1
55.7 66.4 122.1 58.5 68.8 127.3 628 61.8 53.8 68 121.8 56.4 70.4
126.8 648 64.5 50.7 70.6 121.3 52.1 72.4 124.5 665 66.2 48.5 72.1
120.6 52.1 74.9 127 690 67.9 46.2 73.6 119.8 50 76.4 126.4 723 70
43.3 75.3 118.6 47.9 78.4 126.3 732 71.7 40.8 76.7 117.5 45.7 79.9
125.6 763 73.4 38.4 78.1 116.5 41.5 80.4 121.9 809 75.1 35.9 79.5
115.4 38.3 81.4 119.7 825 76.8 33.3 80.8 114.1 36.2 82.9 119.1 847
78.8 30.1 82.4 112.5 34 84.9 118.9 866 80.5 27.5 83.8 111.3 33 86.9
119.9 925 82.3 24.8 85.1 109.9 29.8 87.9 117.7 1010 84.6 20.9 86.9
107.8 26.6 89.9 116.5 1100 87.4 16.3 88.9 105.2 21.3 91.5 112.8
1210 91.1 11.7 92.3 104 17 95 112 1300 93.9 8.7 94.9 103.6 10.6 96
106.6 1600 95.6 6.7 96.5 103.2 10.6 98.5 109.1 2100 97.3 4.8 98.1
102.9 7.4 99.5 106.9 2560 99 2.9 99.7 102.6 3.2 100 103.2
[0495] No interaction was observed between the sTREM-1 level and
age, gender, lactate, cardiac comorbidities, non-cardiac
comorbidities, SOFA, SOFA.4, APACHE2, or SAPS2 (all interactions
p>0.1). sTREM-1 was an independent predictor of 28-day mortality
and provided additional information to all tested covariates (all
p<0.001).
[0496] 28-day and 90-day non-survivors showed sTREM-1 levels that
were significantly elevated at baseline when compared to the
sTREM-1 levels at baseline of 28-day and 90-day survivors (FIG.
3A-B). Interestingly, sTREM-1 values at day 1 were correlated with
sTREM-1 values at day 2 and day 3 (Spearman rank correlation r=0.91
(CI: 0.88, 0.93; p<0.001; n=260) and Spearman rank correlation
r=0.86 (CI: 0.81, 0.9); p<0.001; n=219)), and sTREM-1 values at
day 2 were correlated with sTREM-1 values at day 3 (Spearman r=0.96
(CI: 0.95, 0.97; p<0.001, n=213)). sTREM-1 values at day 2 and
day 3 were still found to be elevated in 28-day and 90-day
non-survivors as compared to the sTREM-1 values at day 2 and day 3
in 28-day and 90-day survivors (FIG. 4A-B and FIG. 5A-B). sTREM-1
values at day 1, day 2 and day 3 were also associated with renal
replacement therapy (RRT) at admission and during the length of
stay (FIG. 6A-C).
[0497] Among all parameters tested, sTREM-1 showed to be better
correlated to SOFA score (Spearman rank correlation r=0.42,
p<0.001; n=254).
[0498] All these results tend to demonstrate that the sTREM-1 level
is associated with severity, renal replacement therapy and
mortality. These results thus confirm that the sTREM-1 level may be
used as a reliable biomarker for the identification of patients at
risk of complicated outcome.
Example 3
[0499] Materials and Methods
[0500] sTREM-1 Measurements: Validated ELISA Method for the
Quantitative Determination of sTREM-1 in Human K.sub.2-EDTA
Plasma
[0501] The quantitative determination of sTREM-1 was carried out
using the method described in Example 2.
[0502] Evaluation of sTREM-1 in a Phase Ha Clinical Trial with
Nangibotide in Septic Shock Patients (NCT03158948)
[0503] This was a European prospective, observational,
multinational study in 14 centers from 4 countries (France,
Belgium, the Netherlands, and Spain). Patients were recruited from
July 2017 to June 2018. The study was conducted in accordance with
the principles set forth in the Declaration of Helsinki and
subsequent amendments, the Guidelines of the International
Conference on Harmonization on GCP, as well as the requirements of
national drug and data protection laws and other applicable
regulatory requirements.
[0504] This was a randomized, double-blind, two-stage,
placebo-controlled study. As described in FIG. 7, the study was
composed of 2 stages with a similar treatment regimen, in which
0.3, 1.0 or 3.0 mg/kg/h of nangibotide (corresponding to peptide
LR12, having an amino acid sequence as set forth in SEQ ID NO: 9,
also known as motrem) was tested versus placebo. All patients with
a diagnosis of septic shock were considered for study
participation. All potential study patients underwent standard care
management procedures. Patients with evidence of septic shock
according to the Sepsis-3 definition (Singer et al, JAMA 2016) were
considered for inclusion (i.e., patients suffering from sepsis and
having (i) persisting hypotension requiring vasopressors to
maintain their mean arterial pressure .gtoreq.65 mm Hg and (ii) a
serum lactate level >2 mmol/L (18 mg/dL) despite adequate volume
resuscitation).
[0505] Exclusion criteria were previous episode of septic shock
(vasopressor administration) within current hospital stay,
underlying concurrent immunodepression, solid organ transplant
requiring immunosuppressive therapy, known pregnancy, prolonged QT
syndrome (QTc .gtoreq.440 ms), shock of any other cause, e.g.,
hypotension related to gastrointestinal bleeding, ongoing
documented or suspected endocarditis, history of prosthetic heart
valves, end-stage neurological disease, end-stage cirrhosis (Child
Pugh Class C), acute Physiology And Chronic Health Evaluation
(APACHE) II score .gtoreq.34, end stage chronic renal disease
requiring chronic dialysis, home oxygen therapy on a regular basis
for >6 h/day, severe obesity (body mass index [BMI].gtoreq.40),
recent cardiopulmonary resuscitation (within current hospital
stay), moribund patients and decision to limit full care taken
before obtaining informed consent.
[0506] After screening for eligibility, patients were randomized to
one of the treatment arms. Then, they received either a 5 mg/kg
loading dose of nangibotide over 15 minutes followed by a
continuous intravenous (i.v.) infusion of nangibotide or a matching
placebo on top of standard of care. Treatment with study drug was
initiated as early as possible, but no later than 24 hours after
the onset of septic shock, defined by the start of vasopressor
therapy. Patients were treated until 12 (.+-.2) hours after the
resolution of their septic shock (defined as vasopressor
withdrawal) with a maximum treatment duration of 5 days (120
hours).
[0507] Stage 1 was performed to investigate ascending doses of
nangibotide or placebo in a sequential design in cohorts of 4
patients (3:1 randomization). After completion of a cohort, safety
and PK data were reviewed by an independent data safety monitoring
board (DSMB) and the study could progress to the next cohort/stage.
Stage 2 investigated 3 doses of nangibotide in a randomized,
balanced, parallel-group design involving up to 3 doses of
nangibotide and a placebo arm. Only dose arms of nangibotide
considered to be safe and well tolerated during Stage 1 were
administered in Stage 2.
[0508] PK Analytical Method for Phase IIa with Nangibotide
[0509] Blood nangibotide concentrations were determined by a
validated LC-MS/MS method in accordance with European Medicines
Agency (EMA) guidelines on bioanalytical method validation
EMEA/CHMP/EWP/192217/2009, version EMA/275542/2014. The peptide was
stabilized in the blood samples by treatment with a 10%
trichloroacetic acid solution (1500 mg/mL, 4.degree. C.) and
centrifugation (3500 g, 10 min, 4.degree. C.) to collect and freeze
the supernatant before analysis. The lower limit of quantification
(LLOQ) and upper limit of quantification (ULOQ) were 5 ng/mL and
1000 ng/mL respectively. Standard curve was linear over the range 5
ng/mL to 1000 ng/mL with a correlation coefficient greater than
0.995. Within-run and between-run precision and accuracy within
+/-20% LLOQ and +/-15% (other levels up to ULOQ) were achieved
during a 3-run validation for quality controls (QCs) for
nangibotide.
[0510] Immunogenicity Evaluations
[0511] Anti-nangibotide antibodies in serum were evaluated at
baseline, day 8 and day 28, by an indirect ELISA method validated
according to methods described by Shankar et al. (J Pharm Biomed
Anal. 2008 Dec. 15; 48(5):1267-81) and EMA guidance on
immunogenicity assessment of biotechnology-derived therapeutic
proteins (EMEA/CHMP/BMWP/14327/2006 issued on April 2008).
Screening samples determined as positive were further assessed
using the confirmatory assay where the cut point was established to
have 0.1% of false positive samples (Shankar et al., J Pharm Biomed
Anal. 2008 Dec. 15; 48(5):1267-81). Validation of the method showed
no drug interference. The sensitivity of the assay was established
to be 0.3125 .mu.g/mL.
[0512] Statistical Analysis for the Phase IIa with Nangibotide
[0513] Adverse events (AEs) were coded using the MedDRA dictionary
version 18.1. Treatment-emergent adverse events (TEAEs) were
summarized and listed. Analysis was performed in 49 septic shock
patients. The demographic and baseline characteristics were
summarized in a descriptive fashion within each population:
categorical variables were summarized by contingency tables
(frequencies and percentages). sTREM-1 levels at day 1 were also
described (mean, standard deviation, standard error, median,
quartiles and range, minimum and maximum values). Mortality rates
were reported by frequency and percentage. For continuous
endpoints, i.e., the sequential organ failure assessment (SOFA),
missing data were imputed using LOCF (Last Observation
Carry-Forward). For time-to-event analyses, i.e., duration of
invasive mechanical ventilation (IMV), duration of continuous renal
replacement therapy (CRRT), survival, censored data techniques were
utilized. In addition, a sensitivity analysis of time to event data
(except survival) was performed using censoring at maximum limit
for the corresponding observation period (i.e., either day 28 or
day 90). The number of days alive and free from any vasopressor,
IMV (Invasive Mechanical Ventilation) or CRRT (Continuous Renal
Replacement Therapy) use was calculated as the total number of
calendar days from the start of treatment (day 0) to day-28 visit
when a subject is alive and does not require any vasopressor, IMV
or CRRT treatment. The following comparisons were made: each
nangibotide dose arm to placebo, all nangibotide doses combined
versus placebo, and dose-response using an ordinal trend test.
Exploratory composite endpoint: incidence (number and proportion)
of patients alive at day 28 and free of vasopressor, free of IMV
and free of CRRT were summarized by treatment and compared between
treatments with a Fisher's exact test. For pharmacodynamic markers,
log transformed data were represented as percentage change from
baseline at day 5 or End of Infusion (EOI) if end of infusion was
stopped before day 5. High sTREM-1 and low sTREM-1 patients were
defined as patients who displayed a sTREM-1 blood concentration at
entry to ICU (baseline, or day 0) below and above the median of the
population, i.e., 433 pg/mL, respectively. "All nangibotide" refers
to analyses obtained by pooling data from 0.3, 1 and 3 mg/kg/h
nangibotide-treated groups.
[0514] Results
[0515] Patient Disposition and Baseline Characteristics
[0516] 50 patients were randomized and 49 treated (one patient died
before dosing). As shown in FIG. 8, twelve patients were randomized
in each group and one additional patient was added to the 0.3
mg/kg/h group. All discontinuations were due to death. Thirty-six
patients survived until day 90. All groups were well-balanced in
terms of baseline characteristics (age, gender, race, weight,
height, and BMI not shown). Average age was 64 (.+-.13). On
average, patients were under vasopressors for 15 hours (ranging
from 3 h to 24 h) before starting the treatment with no significant
differences between groups; 15 patients out of 49 were treated
after receiving vasopressors between 3 and 12h (30.6%). The
demographics and general baseline characteristics of the patients
are depicted in Table 3 below.
TABLE-US-00003 TABLE 3 Demographics and baseline characteristics of
patients of the phase IIa with nangibotide Placebo 0.3 mg/kg/h 1.0
mg/kg/h 3.0 mg/kg/h Total N = 12 N = 13 N = 12 N = 12 N = 49
Primary type of infection at screening, number (%) Bacterial
Gram-positive 2 (16.7%) 5 (38.5%) 5 (41.7%) 6 (50.0%) 18 (36.7%)
Bacterial Gram-negative 4 (33.3%) 5 (38.5%) 6 (50.0%) 1 (8.3%) 16
(32.7%) Viral -- -- -- 1 (8.3%) 1 (2.0%) Unknown.sup.1 5 (41.7%) 3
(23.1%) -- 2 (16.7%) 10 (20.4%) Other.sup.2 1 (8.3%) -- 1 (8.3%) 2
(16.7%) 4 (8.2%) Site of infection, number (%) Abdominal cavity 5
(41.7%) 8 (61.5%) 4 (33.3%) 4 (33.3%) 21 (42.9%) Lung 6 (50.0%) 5
(38.5%) 6 (50.0%) 8 (66.7%) 25 (51.0%) Urinary tract 1 (8.3%) -- 2
(16.7%) -- 3 (6.1%) Vasopressor duration, 16.4 (6.3) 16.9 (6.5)
14.2 (4.8) 13.4 (5.9) 15.3 (5.9) h, mean (SD) IMV, number (%) 9
(75.0%) 10 (76.9%) 11 (91.7%) 9 (75%) 39 (79.6%) CRRT, number (%)
-- 2 (15.4%) 2 (16.7%) 1 (8.3%) 5 (10.2%) APACHE II, mean (SD) 22.3
(7.8) 22.5 (4.9) 25.1 (9.2) 24.4 (5.6) 23.6 6.9) SOFA, mean (SD)
10.1 (3.6) 9.7 (3.0) 10.9 (3.6) 10.2 (2.6) 10.2 (3.) Lactate.sup.3,
mmol/L, mean (SD) 3.9 (2.5) 4.0 (2.4) 4.3 (1.8) 3.6 (1.4) 4.0 (2.0)
NED.sup.3, .mu.g/kg/min, mean (SD) 0.4 (0.4) 0.4 (0.6) 0.4 (0.3)
0.8 (0.6) 0.5 (0.5) .sup.1From unknown primary type of infections
(10 patients), during the study 4 patients got negative cultures (3
in placebo and 1 in 3 mg/kg/h); 3 had mixed infections (1 in
placebo, 1 in 0.3 mg/kg/h and 1 in 3 mg/kg/h); 1 had a
gram-positive infection (placebo); and 2 had a gram-negative
infection (2 in 0.3 mg/kg/h). .sup.2Two correspond to 1 viral and
gram-negative infection (placebo); 1 viral and gram-positive
infection (1 mg/kg/h); 1 mixed gram-positive and gram-negative
infection (3 mg/kg/h) and 1 misclassified patient having a
gram-positive infection (3 mg/kg/h). .sup.3Data from the Clinical
Coordinating Center data base. IMV: invasive mechanical
ventilation; CRRT: continuous renal replacement therapy; NED:
norepinephrine equivalent dose.
[0517] Safety
[0518] No adverse events (AEs) led to treatment withdrawal. The
number of serious adverse events/adverse events (SAEs/AEs) and
number of patients with SAEs/AEs was comparable between the
nangibotide groups and the placebo group. Proportionally, the
number of SAEs and patients with SAEs was higher in the placebo
group. The number of AEs and patients with AEs with a severe
intensity was comparable between treatment groups but
proportionally higher in the placebo group. The numbers of AEs by
system organ classes were comparable between treatment groups. AEs
related to infections and cardiac disorders were proportionally
higher in the nangibotide groups. The most frequent (>10%) AEs
were atrial fibrillation, anemia, pleural effusion and
thrombocytopenia. The incidence of atrial fibrillation and
thrombocytopenia was proportionally higher in the nangibotide
groups. Two suspected unexpected serious adverse reactions (SUSARs)
without obvious explanation were reported and they were both in the
nangibotide-treated group. There was no definitive explanation
pointing towards relation to nangibotide treatment. As shown in
Table 4 below, nangibotide was safe and well tolerated up to 3
mg/kg/h.
TABLE-US-00004 TABLE 4 Treatment emergent adverse events (TEAEs)
0.3 1.0 3.0 Placebo mg/kg/h mg/kg/h mg/kg/h Total All TEAEs N = 61
N = 56 N = 56 N = 61 N = 234 All TEAEs with 17 12 13 8 50 intensity
severe (27.9%) (21.4%) (23.2%) (13.1%) (21.4%) All TEAEs related to
4 0 4 0 8 study drug (by (6.6%) (0.0%) (7.1%) (0.0%) (3.4%)
investigator) All serious TEAEs 9 5 3 5 22 (including death)
(14.8%) (8.9%) (5.4%) (8.2%) (9.4%) All TEAEs leading to 0 0 0 0 0
treatment withdrawal (0.0%) (0.0%) (0.0%) (0.0%) (0.0%)
[0519] Pharmacokinetics
[0520] For all subjects in all groups, the pre-dose sample (blood
sample withdrawn before the start of nangibotide infusion, i.e.,
baseline sample) did not contain quantifiable nangibotide. Mean
nangibotide blood concentrations over time obtained from each group
(i.e., patients who received 0.3 mg/kg/h, patients who received 1.0
mg/kg/h and who received 3.0 mg/kg/h) from the start of the
intravenous infusion are shown in FIG. 9A. Nangibotide showed a
linear behavior from the start to the end of intravenous infusion.
Nangibotide blood concentration was found to be proportional with
the dose. After the end of infusion, the product was quickly
cleared (FIG. 9B) and showed a similar pattern of pharmacokinetics
similar to that previously observed in healthy volunteers. No drug
was detectable in blood 2 hours after the end of infusion,
therefore no persistent pharmacological effect is foreseen after
this period.
[0521] Mortality
[0522] There was a consistent trend in the proportion of death
patients in favor of nangibotide-treated groups versus placebo.
Indeed, all-cause mortality at day-28 was 14% ( 5/37) in pooled
nangibotide groups and 25% ( 3/12) in placebo group (see Table 5
below).
TABLE-US-00005 TABLE 5 Mortality in the whole population
Nangibotide Nangibotide Nangibotide Nangibotide Placebo 0.3 mg/kg/h
0.3 mg/kg/h 0.3 mg/kg/h pooled (N = 12) (N = 13) (N = 12) (N = 12)
(N = 37) All-cause Number 3 (25%) 2 (15%) 1 (8.3%) 3 (25%) 6 (16%)
mortality of death at 28 days (%) p-value 0.5483 0.2613 1.0000
0.5273 vs. Placebo All-cause Number 2 (17%) 1 (7.7%) 1 (8.3%) 2
(17%) 4 (11%) mortality of death at 5 days (%) p-value 0.4919
0.5340 1.0000 0.6230 vs. Placebo All-cause Number 3 (25%) 4 (32%) 2
(17%) 4 (33%) 10 (27%) mortality of death at 90 days (%)
[0523] In the subgroup with sTREM-1 levels at baseline above the
median, i.e., above 433 pg/mL, all deaths were related to sepsis.
In this group, all deaths occurred during the first week of the
study. The day-5 mortality was calculated as 40% ( ) and 20% (
4/20) in placebo and nangibotide groups respectively (see Table 6
below).
TABLE-US-00006 TABLE 6 Mortality in the subgroup with sTREM-1
levels at baseline above the median (433 pg/mL) Nangibotide
Nangibotide Nangibotide Nangibotide Placebo 0.3 mg/kg/h 0.3 mg/kg/h
0.3 mg/kg/h pooled (N = 5) (N = 6) (N = 6) (N = 8) (N = 20) Sepsis-
Number 2 (40%) 1 (17%) 1 (17%) 2 (25%) 4 (20%) related of death
mortality (%) at 28 days p-value 0.3819 0.3819 0.5748 0.3982 vs
Placebo Sepsis- Number 2 (40%) 1 (17%) 1 (17%) 2 (25%) 4 (20%)
related of death mortality (%) at 5 days p-value 0.3819 0.3819
0.5748 0.3982 vs Placebo Sepsis- Number 2 (40%) 1 (17%) 1 (17%) 2
(25%) 4 (20%) related of death mortality (%) at 90 days p-value
0.3819 0.3819 0.5748 0.3982 vs Placebo
[0524] Organ Failure Parameters
[0525] Although no statistically significant differences were seen
in clinical parameters between nangibotide-treated groups in
comparison to the placebo group, a consistent numerical trend in
favor of nangibotide was observed on SOFA score decrease during the
first 5 days (additional decrease in treated groups versus placebo
group). For example, as shown in Table 7 below, the difference in
SOFA score in the "all nangibotide" group as compared to the
"placebo" group was -0.70 (95% CI: -2.41, 1.01), ranging from -1.66
(95% CI: -3.73, 0.40) to 0.34 (95% CI: -1.76, 2.44) from 0.3 to 3
mg/kg/h doses.
TABLE-US-00007 TABLE 7 SOFA score evolution and difference between
treatment arms and placebo in the whole population Comparison vs.
placebo Estimated 95% Confidence p-value vs. p-value dose- Day
Treatment LSMean (SE) difference (SE) interval placebo response Day
1 Placebo -0.92 (0.54) Nangibotide 0.3 mg/kg/h -0.65 (0.52) 0.26
(0.75) (-1.25, 1.78) 0.7273 Nangibotide 1.0 mg/kg/h -0.39 (0.54)
0.53 (0.77) (-1.01, 2.07) 0.4926 Nangibotide 3.0 mg/kg/h -0.33
(0.54) 0.59 (0.76) (-0.95, 2.13) 0.4454 All nangibotide -0.45
(0.31) 0.46 (0.62) (-0.79, 1.71) 0.4628 0.4049 Day 2 Placebo -1.33
(0.65) Nangibotide 0.3 mg/kg/h -2.27 (0.63) -0.94 (0.91) (-2.76,
0.89) 0.3079 Nangibotide 1.0 mg/kg/h -1.97 (0.65) -0.64 (0.92)
(-2.50, 1.22) 0.4941 Nangibotide 3.0 mg/kg/h -0.91 (0.65) 0.42
(0.92) (-1.44, 2.28) 0.6497 All nangibotide -1.72 (0.37) -0.38
(0.75) (-1.90, 1.13) 0.6118 0.5942 Day 3 Placebo -1.25 (0.70)
Nangibotide 0.3 mg/kg/h -2.67 (0.68) -1.42 (0.98) (-3.40, 0.55)
0.1534 Nangibotide 1.0 mg/kg/h -1.87 (0.71) -0.62 (1.00) (-2.63,
1.39) 0.5363 Nangibotide 3.0 mg/kg/h -0.91 (0.70) 0.34 (1.00)
(-1.67, 2.35) 0.7337 All nangibotide -1.82 (0.40) -0.57 (0.81)
(-2.20, 1.07) 0.4872 0.5652 Day 4 Placebo -1.41 (0.69) Nangibotide
0.3 mg/kg/h -2.91 (0.67) -1.50 (0.96) (-3.44, 0.45) 0.1280
Nangibotide 1.0 mg/kg/h -1.95 (0.70) -0.53 (0.98) (-2.51, 1.45)
0.5912 Nangibotide 3.0 mg/kg/h -1.16 (0.69) 0.26 (0.98) (-1.72,
2.23) 0.7931 All nangibotide -2.00 (0.40) -0.59 (0.80) (-2.20,
1.02) 0.4640 0.5772 Day 5 Placebo -1.25 (0.74) Nangibotide 0.3
mg/kg/h -2.91 (0.71) -1.66 (1.03) (-3.73, 0.40) 0.1115 Nangibotide
1.0 mg/kg/h -2.03 (0.74) -0.78 (1.04) (-2.88, 1.32) 0.4594
Nangibotide 3.0 mg/kg/h -0.90 (0.74) 0.34 (1.04) (-1.76, 2.44)
0.7441 All nangibotide -1.95 (0.42) -0.70 (0.85) (-2.41, 1.01)
0.4333 0.5645
[0526] Interestingly, these differences were more important in the
baseline high sTREM-1 subgroup (patients with a sTREM-1 level at
baseline above the median, i.e., above 433 pg/mL). Indeed, as shown
in Table 8 below, the difference in SOFA score in the "all
nangibotide" group as compared to the "placebo" group was -1.5 (95%
CI: -3.83, 0.84), ranging from -2.85 (95% CI: -5.68, -0.02;
p=0.0487) to -0.16 (95% CI: -2.80, 2.48) from 0.3 to 3 mg/kg/h
doses.
TABLE-US-00008 TABLE 8 SOFA score evolution and difference between
treatment arms and placebo in patients with high sTREM-1 at
baseline Comparison vs. placebo Estimated 95% Confidence p-value
vs. p-value Day Treatment LSMean (SE) difference (SE) interval
placebo dose-response Day 1 Placebo 0.02 (0.66) Nangibotide 0.3
mg/kg/h 0.16 (0.60) 0.14 (0.90) (-1.73, 2.01) 0.8793 Nangibotide
1.0 mg/kg/h -0.50 (0.59) -0.52 (0.89) (-2.37, 1.33) 0.5655
Nangibotide 3.0 mg/kg/h -0.38 (0.51) -0.40 (0.84) (-2.14, 1.35)
0.6415 All nangibotide -0.24 (0.33) -0.26 (0.74) (-1.80, 1.29)
0.7306 0.4930 Day 2 Placebo -0.37 (0.89) Nangibotide 0.3 mg/kg/h
-1.52 (0.80) -1.14 (1.21) (-3.66, 1.38) 0.3566 Nangibotide 1.0
mg/kg/h -2.00 (0.80) -1.63 (1.19) (-4.12, 0.87) 0.1887 Nangibotide
3.0 mg/kg/h -0.50 (0.69) -0.13 (1.13) (-2.49, 2.23) 0.9094 All
nangibotide -1.34 (0.44) -0.97 (1.00) (-3.05, 1.12) 0.3450 0.8084
Day 3 Placebo -0.09 (0.98) Nangibotide 0.3 mg/kg/h -2.73 (0.89)
-2.64 (1.34) (-5.43, 0.16) 0.0631 Nangibotide 1.0 mg/kg/h -2.00
(0.89) -1.90 (1.32) (-4.67, 0.86) 0.1655 Nangibotide 3.0 mg/kg/h
-0.39 (0.77) -0.30 (1.25) (-2.91, 2.31) 0.8128 All nangibotide
-1.71 (0.49) -1.61 (1.11) (-3.92, 0.69) 0.1601 0.9662 Day 4 Placebo
-0.52 (1.03) Nangibotide 0.3 mg/kg/h -3.22 (0.94) -2.70 (1.41)
(-5.63, 0.24) 0.0697 Nangibotide 1.0 mg/kg/h -1.83 (0.93) -1.31
(1.39) (-4.21, 1.59) 0.3558 Nangibotide 3.0 mg/kg/h -0.51 (0.81)
0.00 (1.31) (-2.74, 2.75) 0.9976 All nangibotide -1.85 (0.52) -1.34
(1.16) (-3.76, 1.09) 0.2639 0.7397 Day 5 Placebo -0.35 (1.00)
Nangibotide 0.3 mg/kg/h -3.20 (0.90) -2.85 (1.36) (-5.68, -0.02)
0.0487 Nangibotide 1.0 mg/kg/h -1.83 (0.90) -1.48 (1.34) (-4.28),
1.31) 0.2813 Nangibotide 3.0 mg/kg/h -0.51 (0.78) -0.16 (1.27)
(-2.80, 2.48) 0.9007 All nangibotide -1.85 (0.50) -1.50 (1.12)
(-3.83, 0.84) 0.1963 0.8270
[0527] Conversely, these differences were less important in the
baseline low sTREM-1 subgroup (patients with a sTREM-1 level at
baseline below the median, i.e., below 433 pg/mL). Indeed, as shown
in Table 9 below, the difference in SOFA score in the "all
nangibotide" group as compared to the "placebo" group at day 5 was
-0.18 (95% CI: -2.95, 2.59), ranging from -0.71 (95% CI: -3.97,
2.54) to 0.12 (95% CI: -3.70, 3.94) from 0.3 to 3 mg/kg/h
doses.
TABLE-US-00009 TABLE 9 SOFA score evolution and difference between
treatment arms and placebo in patients with low sTREM-1 at baseline
Comparison vs. placebo Estimated 95% Confidence p-value vs. p-value
Day Treatment LSMean (SE) difference (SE) interval placebo
dose-response Day 1 Placebo -1.68 (0.80) Nangibotide 0.3 mg/kg/h
-1.40 (0.80) 0.29 (1.13) (-2.09, 2.66) 0.8036 Nangibotide 1.0
mg/kg/h -0.01 (0.88) 1.67 (1.20) (-0.85, 4.19) 0.1814 Nangibotide
3.0 mg/kg/h -0.35 (1.06) 1.33 (1.33) (-1.45, 4.11) 0.3285 All
nangibotide -.059 (0.53) 1.10 (0.96) (-0.92, 3.11) 0.2692 0.2117
Day 2 Placebo -2.13 (0.92) Nangibotide 0.3 mg/kg/h -2.99 (0.92)
-0.86 (1.30) (-3.58, 1.87) 0.5181 Nangibotide 1.0 mg/kg/h -1.62
(1.02) 0.51 (1.38) (-2.39, 3.40) 0.7189 Nangibotide 3.0 mg/kg/h
-1.87 (1.22) 0.26 (1.53) (-2.93, 3.46) 0.8653 All nangibotide -2.16
(0.61) -0.03 (1.11) (-2.35, 2.29) 0.9789 0.6582 Day 3 Placebo -2.11
(1.02) Nangibotide 0.3 mg/kg/h -2.68 (1.02) -0.57 (1.44) (-3.59,
2.45) 0.6963 Nangibotide 1.0 mg/kg/h -1.51 (1.13) 0.61 (1.53)
(-2.60, 3.81) 0.6971 Nangibotide 3.0 mg/kg/h -2.10 (1.35) 0.01
(1.69) (-3.53, 3.55) 0.9950 All nangibotide -2.10 (0.67) 0.02
(1.23) (-2.55, 2.58) 0.9903 0.8219 Day 4 Placebo -2.13 (0.94)
Nangibotide 0.3 mg/kg/h -2.70 (0.94) -0.57 (1.32) (-3.34, 2.20)
0.6709 Nangibotide 1.0 mg/kg/h -1.79 (1.03) 0.34 (1.41) (-2.61,
3.29) 0.8122 Nangibotide 3.0 mg/kg/h -2.62 (1.24) -0.49 (1.55)
(-3.74, 2.76) 0.7569 All nangibotide -2.37 (0.62) -0.24 (1.13)
(-2.60, 2.12) 0.8335 0.9109 Day 5 Placebo -1.99 (1.10) Nangibotide
0.3 mg/kg/h -2.71 (1.10) -0.71 (1.56) (-3.97, 2.54) 0.6513
Nangibotide 1.0 mg/kg/h -1.93 (1.21) 0.06 (1.65) (-3.40, 3.53)
0.9692 Nangibotide 3.0 mg/kg/h -1.87 (1.46) 0.12 (-1.82) (-3.70,
3.94) 0.9480 All nangibotide -2.17 (0.73) -0.18 (1.32) (-2.95,
2.59) 0.8954 0.8441
[0528] Unexpectedly, as shown on FIG. 10, an inverse dose response
was observed, with a more pronounced difference in the low dose
group 0.3 mg/kg/h (-2.85, p=0.0487) as compared to the high dose
group 3 mg/kg/h (-0.16, p=0.9007).
[0529] These results show that the overall treatment-related
decrease in SOFA score observed in the whole population was driven
by a decrease in SOFA score in patients with the highest sTREM-1
baseline values (above the median, i.e., above 433 pg/mL), and no
in-between group was observed in groups of patients with low
baseline sTREM-1 values (below the median, i.e., below 433
pg/mL).
[0530] Similarly, although no statistically significant differences
were seen in clinical parameters between nangibotide-treated groups
in comparison to the placebo group, a consistent numerical trend in
favor of nangibotide was observed on vasopressor use, on invasive
mechanical ventilation (IMV) use and on renal replacement therapy
(RRT) use.
[0531] In the baseline low sTREM-1 subgroup, the difference in the
number of vasopressor free days alive in the "all nangibotide"
group as compared to the "placebo" group was 0.64 (95% CI: -5.78,
7.07; p=0.8368), ranging from 2.29 (95% CI: -5.30, 9.87; p=0.5369)
to 0.07 (95% CI: -8.83, 8.97; p=0.9868) from 0.3 to 3 mg/kg/h
doses. In the baseline high sTREM-1 subgroup, the difference in the
number of vasopressor free days alive in the "all nangibotide"
group as compared to the "placebo" group was 2.94 (95% CI: -9.36,
15.25; p=0.6243), ranging from 4.07 (95% CI: -10.81, 18.94;
p=0.5757) to -0.48 (95% CI: -14.48, 13.53; p=0.9444) from 0.3 to 3
mg/kg/h doses.
[0532] In the whole population, the difference in the number of IMV
(invasive mechanical ventilation) free days alive in the "all
nangibotide" group as compared to the "placebo" group was 1.33 (95%
CI: -6.20, 8.89; p=0.7240), ranging from 2.32 (95% CI: -6.75,
11.40; p=0.6091) to -2.08 (95% CI: -11.34, 7.17; p=0.6524) from 0.3
to 3 mg/kg/h doses. In the baseline low sTREM-1 subgroup, the
difference in the number of IMV (invasive mechanical ventilation)
free days alive in the "all nangibotide" group as compared to the
"placebo" group was -1.30 (95% CI: -9.92, 7.32; p=0.7568), ranging
from 3 (95% CI: -7.18, 13.18; p=0.5455) to -6.32 (95% CI: -18.25,
5.61; p=0.2823) from 0.3 to 3 mg/kg/h doses. In the baseline high
sTREM-1 subgroup, the difference in the number of IMV (invasive
mechanical ventilation) free days alive in the "all nangibotide"
group as compared to the "placebo" group was 5.63 (95% CI: -7.24,
18.51; p=0.3732), ranging from 2.63 (95% CI: -12.93, 18.20;
p=0.7285) to 4.30 (95% CI: -10.35, 18.95; p=0.5483) from 0.3 to 3
mg/kg/h doses.
[0533] In the whole population, the difference in the number of
CRRT (continuous renal replacement therapy) free days alive in the
"all nangibotide" group as compared to the "placebo" group was 2.70
(95% CI: -4.46, 9.86; p=0.4516), ranging from 4.10 (95% CI: -4.53,
12.73; p=0.3435) to 1.50 (95% CI: -7.30, 10.30; p=0.7330) from 0.3
to 3 mg/kg/h doses. In the baseline low sTREM-1 subgroup, the
difference in the number of CRRT (continuous renal replacement
therapy) free days alive in the "all nangibotide" group as compared
to the "placebo" group was 0.77 (95% CI: -4.72, 6.25; p=0.7738),
ranging from 1.43 (95% CI: -5.04, 7.90; p=0.6502) to 0.89 (95% CI:
-6.70, 8.48; p=0.8086) from 0.3 to 3 mg/kg/h doses. In the baseline
high sTREM-1 subgroup, the difference in the number of CRRT
(continuous renal replacement therapy) free days alive in the "all
nangibotide" group as compared to the "placebo" group was 7.85 (95%
CI: -5.22, 20.91; p=0.2254), ranging from 8.67 (95% CI: -7.13,
24.46; p=0.2666) to 7.37 (95% CI: -7.49, 22.24; p=0.3140) from 0.3
to 3 mg/kg/h doses.
[0534] As shown in FIG. 11, these trends were magnified in patients
with high levels of circulating sTREM-1 at baseline (above the
median, i.e., above 433 pg/mL). In addition, in the "all
nangibotide" group, the proportion of patients who were alive and
free of any medical support at day 28 was 70% versus 40% in the
"placebo" group.
[0535] Markers
[0536] A trend towards a decrease during the treatment in
circulating levels of endothelial injury markers (Ang-2, VCAM-1,
VGEFR-1 and E-selectin), inflammatory markers (IL-8, IL-10, MCP-1,
TNF-.alpha. and IL-10) and sTREM-1 was observed in
nangibotide-treated patients, being more pronounced in patients
with a high sTREM-1 level at baseline for Ang-2, IL-6 and sTREM-1.
For example, when no apparent treatment effect was observed on the
Ang-2 level in the whole population, a trend toward a more
pronounced decrease was observed in the "G2" subgroup (i.e.,
patients with baseline sTREM-1 above the median 433 pg/mL) as
compared to the whole population "all" and the "G1" subgroup (i.e.,
patients with baseline sTREM-1 below the median 433 pg/mL) (see
FIG. 12). As shown on FIG. 13, the same trend was observed for the
IL-6 level.
[0537] sTREM-1 as an Efficacy Marker
[0538] Besides its value as an inclusion biomarker, sTREM-1 was
also evaluated as a marker for nangibotide efficacy in this phase
IIa study. The impact of nangibotide treatment on the kinetics of
sTREM-1 over time were evaluated. As shown in FIG. 14A, there were
no apparent changes over time observed in the whole population,
however a trend towards a decrease in sTREM-1 was observed in the
subgroup of patients with high baseline sTREM-1 values.
Interestingly, patients with the highest sTREM-1 change displayed
the highest SOFA decrease at day 5/EOI (end of infusion) (FIG.
14B).
CONCLUSION
[0539] Nangibotide was shown to be safe and well-tolerated in
septic shock patients. In this small exploratory study, a
non-significant lower mortality was observed in the nangibotide
group. Strikingly, clinical parameters and mortality showed a
clearer tendency towards a beneficial effect of nangibotide in
patients with a high circulating level of sTREM-1 at baseline.
[0540] Highly variable and controversial results have been
published about the use of sTREM-1 as a prognostic marker in septic
shock patients. This may be partly due to the lack of validated and
reproducible quantitative assay. We have validated such a method,
in accordance with the European Medicines Agency guideline:
Committee for Medicinal Products for Human Use (CHMP) (2011),
Guideline on bioanalytical method validation
EMEA/CHMP/EWP/192217/2009 Rev. 1 Corr. 2. The validated method is
described in the hereinabove (see Materials and Methods), and was
used for the retrospective measurements of sTREM-1 levels in the
two independent clinical cohorts (AdrenOSS cohort and the cohort
from the phase IIa clinical study with nangibotide).
[0541] Interestingly, as shown in Table 10 below, comparable data
were obtained in these two independent European cohorts of 293 and
49 patients.
TABLE-US-00010 TABLE 10 sTREM-1 general descriptive in the AdrenOSS
cohort and in the phase IIa clinical trial with nangibotide
AdrenOSS Samples Median Q1 Q2 Q3 Q4 sTREM-1 June 2015 497 226 372
635 1110 (pg/mL) to May 2016 [42, 299] [300, 497] [500, 809] [810,
5540] [min, max] N = 293 patients All-Cause Mortality at 28 days
(%) 12 30 37 49 Phase IIa nangibotide Samples Median Q1 Q2 Q3 Q4
sTREM-1 July 2017 to 433 250 354 552 990 (pg/mL) March 2018 [154,
302] [304, 412] [433, 693] [719, 1960] [min, max] N = 49 patients
All-Cause Mortality at 28 days (%) 8 17 8 42
[0542] sTREM-1 values at baseline were associated with 28-day
mortality: 12% and 8% mortality rates were observed in sTREM-1
quartile Q1 subgroup, and 49% and 42% mortality rates were observed
in sTREM-1 quartile Q4 subgroup. The median values were 497 pg/mL
in the AdrenOSS cohort, and 433 pg/mL in the phase IIa with
nangibotide, with min/max values (in pg/mL) at 42/5540 and 154/1960
pg/mL, respectively, which suggest a similar distribution between
the two cohorts.
[0543] These results confirm the need for a validated method for
the collection of reliable data between different cohorts. They
also confirm that, with such a method, a reference value from a
reference population can be used for the derivation of a
predetermined sTREM-1 value, which can be used in further
studies.
[0544] Overall, this study aimed to evaluate the value of sTREM-1
level assessment for patient selection before nangibotide
administration. Indeed, discordant data from the literature showed
that high sTREM-1 values at baseline were associated with
complicated outcome (Charles et al., BMC Infect Dis. 2016 Oct. 12;
16(1):559) despite previous clinical data where low sTREM-1 values
at baseline were associated with complicated outcome (Gibot et al.,
Crit Care Med. 2005 April; 33(4):792-6.). It therefore remained
unclear whether sTREM-1 values were associated with severity and
clinical outcome. The mechanism of sTREM-1 release was also
unclear.
[0545] In this study, we demonstrated for the first time that
sTREM-1 release depends on TREM-1 activation. Therefore, sTREM-1
may be used as a reliable surrogate of TREM-1 pathway activation.
It may thus allow to identify the patients who will most likely
benefit from an anti-TREM-1 therapeutic approach such as
nangibotide (corresponding to peptide LR12, having an amino acid
sequence as set forth in SEQ ID NO: 9).
[0546] We also have demonstrated, with a validated method in
accordance with the guidelines, that high sTREM-1 values were
correlated with severity and mortality in septic shock patients,
using two independent cohorts. Therefore, we confirmed that sTREM-1
is a reliable biomarker of severity, and may allow the
identification of patients at risk of complicated outcome. Indeed,
baseline sTREM-1 blood concentration, i.e., within 24 hours
following the diagnostic of septic shock, was found to be
correlated with severity.
[0547] We also performed a subgroup analysis in the phase IIa
clinical trial assessing the safety and tolerability of nangibotide
in septic shock patients, in which the efficacy of nangibotide was
compared between patients with baseline sTREM-1 values below a
predetermined sTREM-1 value, herein defined as the median, and
patients with baseline sTREM-1 values above this predetermined
sTREM-1 value. We confirmed that nangibotide efficacy was more
pronounced in patients with baseline sTREM-1 above the
predetermined sTREM-1 value, i.e., above the median (above 433
pg/mL). This was observed on pharmacodynamic parameters
(endothelium and inflammatory biomarkers), on the use of medical
support (IMV, CRRT, vasopressors), and on the SOFA score.
[0548] The lower pharmacological activity of nangibotide in
patients with a baseline sTREM-1 level below the predetermined
sTREM-1 value cannot be explained by toxicity and side effects,
since nangibotide was well-tolerated at all doses tested, and no
adverse events related to nangibotide treatment were observed at
the highest dose. No anti-nangibotide antibodies were found in the
serum of patients, which also indicate that this apparent lower
pharmacological activity in these patients cannot be related to the
presence of anti-nangibotide neutralizing antibodies that would
have been able to inhibit nangibotide. The blood concentrations of
nangibotide were proportional with the dose, and within the same
range previously observed in healthy volunteers, meaning that there
was no abnormal pharmacokinetic profile of nangibotide in patients
that may have also explained the in-between groups differences. The
product was quickly cleared after the end of infusion, which also
means that no persistent pharmacological activity may explain these
differences.
[0549] Overall, these results confirm that sTREM-1 is a reliable
companion diagnostic marker for patient inclusion and for TREM-1
inhibitor (e.g., nangibotide) administration.
Sequence CWU 1
1
191234PRTHomo sapiens 1Met Arg Lys Thr Arg Leu Trp Gly Leu Leu Trp
Met Leu Phe Val Ser1 5 10 15Glu Leu Arg Ala Ala Thr Lys Leu Thr Glu
Glu Lys Tyr Glu Leu Lys 20 25 30Glu Gly Gln Thr Leu Asp Val Lys Cys
Asp Tyr Thr Leu Glu Lys Phe 35 40 45Ala Ser Ser Gln Lys Ala Trp Gln
Ile Ile Arg Asp Gly Glu Met Pro 50 55 60Lys Thr Leu Ala Cys Thr Glu
Arg Pro Ser Lys Asn Ser His Pro Val65 70 75 80Gln Val Gly Arg Ile
Ile Leu Glu Asp Tyr His Asp His Gly Leu Leu 85 90 95Arg Val Arg Met
Val Asn Leu Gln Val Glu Asp Ser Gly Leu Tyr Gln 100 105 110Cys Val
Ile Tyr Gln Pro Pro Lys Glu Pro His Met Leu Phe Asp Arg 115 120
125Ile Arg Leu Val Val Thr Lys Gly Phe Ser Gly Thr Pro Gly Ser Asn
130 135 140Glu Asn Ser Thr Gln Asn Val Tyr Lys Ile Pro Pro Thr Thr
Thr Lys145 150 155 160Ala Leu Cys Pro Leu Tyr Thr Ser Pro Arg Thr
Val Thr Gln Ala Pro 165 170 175Pro Lys Ser Thr Ala Asp Val Ser Thr
Pro Asp Ser Glu Ile Asn Leu 180 185 190Thr Asn Val Thr Asp Ile Ile
Arg Val Pro Val Phe Asn Ile Val Ile 195 200 205Leu Leu Ala Gly Gly
Phe Leu Ser Lys Ser Leu Val Phe Ser Val Leu 210 215 220Phe Ala Val
Thr Leu Arg Ser Phe Val Pro225 2302150PRTHomo sapiens 2Met Arg Lys
Thr Arg Leu Trp Gly Leu Leu Trp Met Leu Phe Val Ser1 5 10 15Glu Leu
Arg Ala Ala Thr Lys Leu Thr Glu Glu Lys Tyr Glu Leu Lys 20 25 30Glu
Gly Gln Thr Leu Asp Val Lys Cys Asp Tyr Thr Leu Glu Lys Phe 35 40
45Ala Ser Ser Gln Lys Ala Trp Gln Ile Ile Arg Asp Gly Glu Met Pro
50 55 60Lys Thr Leu Ala Cys Thr Glu Arg Pro Ser Lys Asn Ser His Pro
Val65 70 75 80Gln Val Gly Arg Ile Ile Leu Glu Asp Tyr His Asp His
Gly Leu Leu 85 90 95Arg Val Arg Met Val Asn Leu Gln Val Glu Asp Ser
Gly Leu Tyr Gln 100 105 110Cys Val Ile Tyr Gln Pro Pro Lys Glu Pro
His Met Leu Phe Asp Arg 115 120 125Ile Arg Leu Val Val Thr Lys Gly
Phe Arg Cys Ser Thr Leu Ser Phe 130 135 140Ser Trp Leu Val Asp
Ser145 1503225PRTHomo sapiens 3Met Arg Lys Thr Arg Leu Trp Gly Leu
Leu Trp Met Leu Phe Val Ser1 5 10 15Glu Leu Arg Ala Ala Thr Lys Leu
Thr Glu Glu Lys Tyr Glu Leu Lys 20 25 30Glu Gly Gln Thr Leu Asp Val
Lys Cys Asp Tyr Thr Leu Glu Lys Phe 35 40 45Ala Ser Ser Gln Lys Ala
Trp Gln Ile Ile Arg Asp Gly Glu Met Pro 50 55 60Lys Thr Leu Ala Cys
Thr Glu Arg Pro Ser Lys Asn Ser His Pro Val65 70 75 80Gln Val Gly
Arg Ile Ile Leu Glu Asp Tyr His Asp His Gly Leu Leu 85 90 95Arg Val
Arg Met Val Asn Leu Gln Val Glu Asp Ser Gly Leu Tyr Gln 100 105
110Cys Val Ile Tyr Gln Pro Pro Lys Glu Pro His Met Leu Phe Asp Arg
115 120 125Ile Arg Leu Val Val Thr Lys Gly Phe Ser Gly Thr Pro Gly
Ser Asn 130 135 140Glu Asn Ser Thr Gln Asn Val Tyr Lys Ile Pro Pro
Thr Thr Thr Lys145 150 155 160Ala Leu Cys Pro Leu Tyr Thr Ser Pro
Arg Thr Val Thr Gln Ala Pro 165 170 175Pro Lys Ser Thr Ala Asp Val
Ser Thr Pro Asp Ser Glu Ile Asn Leu 180 185 190Thr Asn Val Thr Asp
Ile Ile Arg Tyr Ser Phe Gln Val Pro Gly Pro 195 200 205Leu Val Trp
Thr Leu Ser Pro Leu Phe Pro Ser Leu Cys Ala Glu Arg 210 215
220Met2254250PRTHomo sapiens 4Met Arg Lys Thr Arg Leu Trp Gly Leu
Leu Trp Met Leu Phe Val Ser1 5 10 15Glu Leu Arg Ala Ala Thr Lys Leu
Thr Glu Glu Lys Tyr Glu Leu Lys 20 25 30Glu Gly Gln Thr Leu Asp Val
Lys Cys Asp Tyr Thr Leu Glu Lys Phe 35 40 45Ala Ser Ser Gln Lys Ala
Trp Gln Ile Ile Arg Asp Gly Glu Met Pro 50 55 60Lys Thr Leu Ala Cys
Thr Glu Arg Pro Ser Lys Asn Ser His Pro Val65 70 75 80Gln Val Gly
Arg Ile Ile Leu Glu Asp Tyr His Asp His Gly Leu Leu 85 90 95Arg Val
Arg Met Val Asn Leu Gln Val Glu Asp Ser Gly Leu Tyr Gln 100 105
110Cys Val Ile Tyr Gln Pro Pro Lys Glu Pro His Met Leu Phe Asp Arg
115 120 125Ile Arg Leu Val Val Thr Lys Gly Phe Ser Gly Thr Pro Gly
Ser Asn 130 135 140Glu Asn Ser Thr Gln Asn Val Tyr Lys Ile Pro Pro
Thr Thr Thr Lys145 150 155 160Ala Leu Cys Pro Leu Tyr Thr Ser Pro
Arg Thr Val Thr Gln Ala Pro 165 170 175Pro Lys Ser Thr Ala Asp Val
Ser Thr Pro Asp Ser Glu Ile Asn Leu 180 185 190Thr Asn Val Thr Asp
Ile Ile Arg Glu Lys Ser Met Thr Phe Gly Ile 195 200 205Arg Arg Leu
Asp Val Glu Ser His Pro Leu Pro Pro Leu His Thr Gly 210 215 220His
Phe Arg Ile Ser Gln Phe Phe Ser Gln Ala Gly Thr Gln Ser Leu225 230
235 240His Ser Cys Tyr Lys Gly Lys Pro Thr Pro 245 2505185PRTHomo
sapiens 5Ala Thr Lys Leu Thr Glu Glu Lys Tyr Glu Leu Lys Glu Gly
Gln Thr1 5 10 15Leu Asp Val Lys Cys Asp Tyr Thr Leu Glu Lys Phe Ala
Ser Ser Gln 20 25 30Lys Ala Trp Gln Ile Ile Arg Asp Gly Glu Met Pro
Lys Thr Leu Ala 35 40 45Cys Thr Glu Arg Pro Ser Lys Asn Ser His Pro
Val Gln Val Gly Arg 50 55 60Ile Ile Leu Glu Asp Tyr His Asp His Gly
Leu Leu Arg Val Arg Met65 70 75 80Val Asn Leu Gln Val Glu Asp Ser
Gly Leu Tyr Gln Cys Val Ile Tyr 85 90 95Gln Pro Pro Lys Glu Pro His
Met Leu Phe Asp Arg Ile Arg Leu Val 100 105 110Val Thr Lys Gly Phe
Ser Gly Thr Pro Gly Ser Asn Glu Asn Ser Thr 115 120 125Gln Asn Val
Tyr Lys Ile Pro Pro Thr Thr Thr Lys Ala Leu Cys Pro 130 135 140Leu
Tyr Thr Ser Pro Arg Thr Val Thr Gln Ala Pro Pro Lys Ser Thr145 150
155 160Ala Asp Val Ser Thr Pro Asp Ser Glu Ile Asn Leu Thr Asn Val
Thr 165 170 175Asp Ile Ile Arg Val Pro Val Phe Asn 180
1856175PRTHomo sapiens 6Leu Lys Glu Gly Gln Thr Leu Asp Val Lys Cys
Asp Tyr Thr Leu Glu1 5 10 15Lys Phe Ala Ser Ser Gln Lys Ala Trp Gln
Ile Ile Arg Asp Gly Glu 20 25 30Met Pro Lys Thr Leu Ala Cys Thr Glu
Arg Pro Ser Lys Asn Ser His 35 40 45Pro Val Gln Val Gly Arg Ile Ile
Leu Glu Asp Tyr His Asp His Gly 50 55 60Leu Leu Arg Val Arg Met Val
Asn Leu Gln Val Glu Asp Ser Gly Leu65 70 75 80Tyr Gln Cys Val Ile
Tyr Gln Pro Pro Lys Glu Pro His Met Leu Phe 85 90 95Asp Arg Ile Arg
Leu Val Val Thr Lys Gly Phe Ser Gly Thr Pro Gly 100 105 110Ser Asn
Glu Asn Ser Thr Gln Asn Val Tyr Lys Ile Pro Pro Thr Thr 115 120
125Thr Lys Ala Leu Cys Pro Leu Tyr Thr Ser Pro Arg Thr Val Thr Gln
130 135 140Ala Pro Pro Lys Ser Thr Ala Asp Val Ser Thr Pro Asp Ser
Glu Ile145 150 155 160Asn Leu Thr Asn Val Thr Asp Ile Ile Arg Val
Pro Val Phe Asn 165 170 1757311PRTHomo sapiens 7Met Gly Leu Thr Leu
Leu Leu Leu Leu Leu Leu Gly Leu Glu Gly Gln1 5 10 15Gly Ile Val Gly
Ser Leu Pro Glu Val Leu Gln Ala Pro Val Gly Ser 20 25 30Ser Ile Leu
Val Gln Cys His Tyr Arg Leu Gln Asp Val Lys Ala Gln 35 40 45Lys Val
Trp Cys Arg Phe Leu Pro Glu Gly Cys Gln Pro Leu Val Ser 50 55 60Ser
Ala Val Asp Arg Arg Ala Pro Ala Gly Arg Arg Thr Phe Leu Thr65 70 75
80Asp Leu Gly Gly Gly Leu Leu Gln Val Glu Met Val Thr Leu Gln Glu
85 90 95Glu Asp Ala Gly Glu Tyr Gly Cys Met Val Asp Gly Ala Arg Gly
Pro 100 105 110Gln Ile Leu His Arg Val Ser Leu Asn Ile Leu Pro Pro
Glu Glu Glu 115 120 125Glu Glu Thr His Lys Ile Gly Ser Leu Ala Glu
Asn Ala Phe Ser Asp 130 135 140Pro Ala Gly Ser Ala Asn Pro Leu Glu
Pro Ser Gln Asp Glu Lys Ser145 150 155 160Ile Pro Leu Ile Trp Gly
Ala Val Leu Leu Val Gly Leu Leu Val Ala 165 170 175Ala Val Val Leu
Phe Ala Val Met Ala Lys Arg Lys Gln Gly Asn Arg 180 185 190Leu Gly
Val Cys Gly Arg Phe Leu Ser Ser Arg Val Ser Gly Met Asn 195 200
205Pro Ser Ser Val Val His His Val Ser Asp Ser Gly Pro Ala Ala Glu
210 215 220Leu Pro Leu Asp Val Pro His Ile Arg Leu Asp Ser Pro Pro
Ser Phe225 230 235 240Asp Asn Thr Thr Tyr Thr Ser Leu Pro Leu Asp
Ser Pro Ser Gly Lys 245 250 255Pro Ser Leu Pro Ala Pro Ser Ser Leu
Pro Pro Leu Pro Pro Lys Val 260 265 270Leu Val Cys Ser Lys Pro Val
Thr Tyr Ala Thr Val Ile Phe Pro Gly 275 280 285Gly Asn Lys Gly Gly
Gly Thr Ser Cys Gly Pro Ala Gln Asn Pro Pro 290 295 300Asn Asn Gln
Thr Pro Ser Ser305 310817PRTArtificial SequenceTLT-1 peptide 8Leu
Gln Glu Glu Asp Ala Gly Glu Tyr Gly Cys Met Val Asp Gly Ala1 5 10
15Arg912PRTArtificial SequenceTLT-1 peptide 9Leu Gln Glu Glu Asp
Ala Gly Glu Tyr Gly Cys Met1 5 10106PRTArtificial SequenceTLT-1
peptide 10Leu Gln Glu Glu Asp Ala1 5116PRTArtificial SequenceTLT-1
peptide 11Glu Asp Ala Gly Glu Tyr1 5126PRTArtificial SequenceTLT-1
peptide 12Gly Glu Tyr Gly Cys Met1 51318DNAArtificial
Sequenceprimer 13gtggtgacca aggggttc 181420DNAArtificial
Sequenceprimer 14agatggatgt ggctggaagt 201520DNAArtificial
Sequenceprimer 15gtgaccaagg gtttttcagg 201620DNAArtificial
Sequenceprimer 16acaccggaac cctgatgata 201719DNAArtificial
Sequenceprimer 17aaaggcaaga acgcctgac 191820DNAArtificial
Sequenceprimer 18gggactttac caagagggac 2019107PRTHomo sapiens 19Leu
Lys Glu Gly Gln Thr Leu Asp Val Lys Cys Asp Tyr Thr Leu Glu1 5 10
15Lys Phe Ala Ser Ser Gln Lys Ala Trp Gln Ile Ile Arg Asp Gly Glu
20 25 30Met Pro Lys Thr Leu Ala Cys Thr Glu Arg Pro Ser Lys Asn Ser
His 35 40 45Pro Val Gln Val Gly Arg Ile Ile Leu Glu Asp Tyr His Asp
His Gly 50 55 60Leu Leu Arg Val Arg Met Val Asn Leu Gln Val Glu Asp
Ser Gly Leu65 70 75 80Tyr Gln Cys Val Ile Tyr Gln Pro Pro Lys Glu
Pro His Met Leu Phe 85 90 95Asp Arg Ile Arg Leu Val Val Thr Lys Gly
Phe 100 105
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