U.S. patent application number 15/443674 was filed with the patent office on 2017-06-22 for biochemical analysis of pbmc.
This patent application is currently assigned to TODOS MEDICAL LTD.. The applicant listed for this patent is TODOS MEDICAL LTD.. Invention is credited to Joseph KAPELUSHNIK, Shaul MORDECHAI, Ilana NATHAN, Udi ZELIG, Rami ZIGDON.
Application Number | 20170176327 15/443674 |
Document ID | / |
Family ID | 45067149 |
Filed Date | 2017-06-22 |
United States Patent
Application |
20170176327 |
Kind Code |
A1 |
KAPELUSHNIK; Joseph ; et
al. |
June 22, 2017 |
BIOCHEMICAL ANALYSIS OF PBMC
Abstract
A method is provided comprising, obtaining an infrared (IR)
spectrum of a Peripheral Blood Mononuclear Cells (PBMC) sample by
analyzing the sample by infrared spectroscopy; and based on the
infrared spectrum, generating an output indicative of the presence
of a solid tumor or a pre-malignant condition. Other embodiments
are also provided.
Inventors: |
KAPELUSHNIK; Joseph; (Moshav
Neve Ilan, IL) ; MORDECHAI; Shaul; (Omer, IL)
; NATHAN; Ilana; (Omer, IL) ; ZELIG; Udi;
(Kibbutz Nir Yitzhak, IL) ; ZIGDON; Rami;
(Ra'anana, IL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
TODOS MEDICAL LTD. |
Airport City |
|
IL |
|
|
Assignee: |
TODOS MEDICAL LTD.
Airport City
IL
|
Family ID: |
45067149 |
Appl. No.: |
15/443674 |
Filed: |
February 27, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13701262 |
Feb 12, 2013 |
9606057 |
|
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PCT/IL2011/000426 |
Jun 1, 2011 |
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15443674 |
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61350073 |
Jun 1, 2010 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
G01N 15/1456 20130101;
G01N 2015/1006 20130101; G01N 2021/3595 20130101; G01N 21/35
20130101; G01N 2015/008 20130101; G01N 21/3563 20130101; G01J 3/433
20130101; G01N 21/63 20130101; G01N 2201/12 20130101 |
International
Class: |
G01N 21/3563 20060101
G01N021/3563; G01N 15/14 20060101 G01N015/14 |
Claims
1. A method comprising: obtaining an infrared (IR) spectrum of a
Peripheral Blood Mononuclear Cells (PBMC) sample by analyzing the
sample by infrared spectroscopy; and based on the infrared
spectrum, generating an output indicative of the presence of a
solid tumor or a pre-malignant condition.
2. The method according to claim 1, wherein generating the output
comprises generating the output indicative of the presence of the
solid tumor.
3. The method according to claim 1, wherein analyzing the sample by
infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared (FTIR) spectroscopy, and wherein
obtaining the infrared (IR) spectrum comprises obtaining a Fourier
Transformed Infrared (FTIR) spectrum.
4. The method according to claim 3, wherein analyzing the sample by
infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared microspectroscopy (FTIR-MSP).
5. The method according to claim 1, wherein analyzing comprises
assessing a characteristic of the sample at at least one wavenumber
selected from the group consisting of: 765.+-.4 cm-1, 798.+-.4
cm-1, 809.+-.4 cm-1, 814.+-.4 cm-1, 875.+-.4 cm-1, 997.+-.4 cm-1,
1001.+-.4 cm-1, 1015.+-.4 cm-1, 1103.+-.4 cm-1, 1118.+-.4 cm-1,
1162.+-.4 cm-1, 1221.+-.4 cm-1, 1270.+-.4 cm-1, 1283.+-.4 cm-1,
1295.+-.4 cm-1, 1315.+-.4 cm-1, 1341.+-.4 cm-1, 1367.+-.4 cm-1,
1392.+-.4 cm-1, 1429.+-.4 cm-1, 1440.+-.4 cm-1, 1445.+-.4 cm-1 and
1455.+-.4 cm-1.
6. The method according to claim 5, wherein analyzing comprises
assessing the characteristic at at least two wavenumbers selected
from the group.
7. The method according to claim 5, wherein analyzing comprises
assessing the characteristic at at least three wavenumbers selected
from the group.
8. The method according to claim 5, wherein assessing the
characteristic comprises analyzing a band of the IR spectrum
surrounding at least one wavenumber selected from the group.
9. The method according to claim 1, wherein analyzing the sample
comprises obtaining a second derivative of the infrared (IR)
spectrum of the sample.
10. The method according to claim 1, wherein the infrared (IR)
spectrum includes an absorption spectrum, and wherein obtaining the
infrared (IR) spectrum comprises obtaining the absorption
spectrum.
11. The method according to claim 1, wherein the infrared (IR)
spectrum includes a reflection spectrum, and wherein obtaining the
infrared (IR) spectrum comprises obtaining the reflection
spectrum.
12. The method according to claim 1, wherein generating the output
comprises indicating via the output whether the solid tumor is a
first type of solid tumor or a second type of solid tumor.
13. The method according to claim 1, wherein the solid tumor
includes a solid tumor in tissue selected from the group consisting
of: head and neck, esophagus, and pancreas, and wherein generating
the output comprises generating an output indicative of the
presence of a solid tumor in tissue selected from the group.
14. The method according to claim 1, wherein the solid tumor
includes a solid tumor in tissue selected from the group consisting
of: breast, gastrointestinal tract, prostate, and lung, and wherein
generating the output comprises generating an output indicative of
the presence of a solid tumor in tissue selected from the
group.
15. The method according to claim 14, wherein analyzing comprises
assessing a characteristic of the sample at at least one wavenumber
selected from the group consisting of: 752.+-.4 cm-1, 1030.+-.4
cm-1, 1046.+-.4 cm-1, 1128.+-.4 cm-1, and 1237.+-.4 cm-1, and
wherein generating comprises generating an output indicative of the
presence of a tumor in the breast tissue.
16. The method according to claim 14, wherein analyzing comprises
assessing a characteristic of the sample at at least one wavenumber
selected from the group consisting of: 797.+-.4 cm-1, 830.+-.4
cm-1, 893.+-.4 cm-1, 899.+-.4 cm-1, 1128.+-.4 cm-1, 1298.+-.4 cm-1,
1354 .+-.4 cm-1, 1714.+-.4 cm-1 1725.+-.4 cm-1, 1738,.+-.4 cm-1,
and 3013.+-.4 cm-1, and wherein generating comprises generating an
output indicative of the presence of a tumor in the
gastrointestinal tract tissue.
17. The method according to claim 14, wherein analyzing comprises
assessing a characteristic of the sample at at least one wavenumber
selected from the group consisting of: 765.+-.4 cm-1, 780.+-.4
cm-1, 797.+-.4 cm-1, 851.+-.4 cm-1, 874.+-.4 cm-1, 881.+-.4 cm-1,
913.+-.4 cm-1, 923.+-.4 cm-1, 958.+-.4 cm-1, 968,.+-.4 cm-1,
1044.+-.4 cm-1, 1085.+-.4 cm-1, 1191.+-.4 cm-1, 1241.+-.4 cm-1,
1344.+-.4 cm-1, 1373.+-.4 cm-1, 1417.+-.4 cm-1, 1458.+-.4 cm-1,
1469.+-.4 cm-1, 1692.+-.4 cm-1, 1714.+-.4 cm-1, 1728.+-.4 cm-1,
2852.+-.4 cm-1, and 2984.+-.4 cm, and wherein generating comprises
generating an output indicative of the presence of a tumor in the
lung tissue.
18. The method according to claim 14, wherein analyzing comprises
assessing a characteristic of the sample at at least one wavenumber
selected from the group consisting of: 828.+-.4 cm-1, 932.+-.4
cm-1, 997.+-.4 cm-1, 1059.+-.4 cm-1, 1299.+-.4 cm-1, 1302.+-.4
cm-1, 1403.+-.4 cm-1, 1454.+-.4 cm-1, 1714.+-.4 cm-1, 2979,.+-.4
cm-1, and 3013.+-.4 cm-1, and wherein generating comprises
generating an output indicative of the presence of a tumor in the
prostate tissue.
19. A method comprising: obtaining an infrared (IR) spectrum of a
sample of white blood cells by analyzing the sample by infrared
spectroscopy; and based on the infrared spectrum, generating an
output indicative of the presence of a solid tumor or a
pre-malignant condition.
20. The method according to claim 19, wherein generating the output
comprises generating the output indicative of the presence of the
solid tumor.
21. The method according to claim 19, wherein analyzing the sample
by infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared (FTIR) spectroscopy, and wherein
obtaining the infrared (IR) spectrum comprises obtaining a Fourier
Transformed Infrared (FTIR) spectrum.
22. The method according to claim 21, wherein analyzing the sample
by infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared microspectroscopy (FTIR-MSP).
23. A method for monitoring the effect of an anti-cancer treatment
on a subject undergoing anti-cancer treatment for a solid tumor,
for use with a first Peripheral Blood Mononuclear Cells (PBMC)
sample separated from blood of the subject that was obtained prior
to initiation of the treatment and a second PBMC sample separated
from blood of the subject that was obtained after initiation of the
treatment, the method comprising: obtaining IR spectra of the first
and second PBMC samples by analyzing the first and second PBMC
samples by IR spectroscopy; and based on the IR spectra, generating
an output indicative of the effect of the treatment.
24. The method according to claim 23, wherein analyzing the first
and second PBMC samples by IR spectroscopy comprises analyzing the
samples by Fourier Transformed Infrared spectroscopy, and wherein
obtaining the IR spectra comprises obtaining Fourier Transformed
Infrared (FTIR) spectra.
25. The method according to claim 24, wherein analyzing the first
and second PBMC samples by infrared (IR) spectroscopy comprises
analyzing the first and second PBMC samples by Fourier Transformed
Infrared microspectroscopy (FTIR-MSP).
26. The method according to claim 23, further comprising obtaining
an IR spectrum of a third PBMC sample separated from blood of the
subject that was obtained following termination of the treatment,
by analyzing the third PBMC sample by IR spectroscopy.
27. The method according to claim 3, wherein analyzing comprises
assessing a characteristic of the sample at at least one wavenumber
selected from the group consisting of: 765.+-.4 cm-1, 798.+-.4
cm-1, 809.+-.4 cm-1, 814.+-.4 cm-1, 875.+-.4 cm-1, 997.+-.4 cm-1,
1001.+-.4 cm-1, 1015.+-.4 cm-1, 1103.+-.4 cm-1, 1118.+-.4 cm-1,
1162.+-.4 cm-1, 1221.+-.4 cm-1, 1270.+-.4 cm-1, 1283.+-.4 cm-1,
1295.+-.4 cm-1, 1315.+-.4 cm-1, 1341.+-.4 cm-1, 1367.+-.4 cm-1,
1392.+-.4 cm-1, 1429.+-.4 cm-1, 1440.+-.4 cm-1, 1445.+-.4 cm-1, and
1455.+-.4 cm-1.
28. The method according to claim 27, wherein analyzing comprises
assessing the characteristic at at least two wavenumbers selected
from the group.
29. The method according to claim 27, wherein analyzing comprises
assessing the characteristic at at least three wavenumbers selected
from the group.
30. A method comprising: obtaining an infrared (IR) spectrum of a
Peripheral Blood Mononuclear Cells (PBMC) sample by analyzing the
sample; and based on the infrared spectrum, generating an output
indicative of the presence of a solid tumor or a pre-malignant
condition.
31. The method according to claim 30, wherein generating the output
comprises generating the output indicative of the presence of the
solid tumor.
32. A system for diagnosing a solid tumor, comprising: a data
processor, configured to analyze an infrared (IR) spectrum of a
Peripheral Blood Mononuclear Cells (PBMC) sample of a subject; and
an output unit, configured to generate an output indicative of the
presence of a solid tumor, based on the infrared (IR) spectrum.
33. The system according to claim 32, wherein the data processor is
configured to calculate a second derivative of the infrared (IR)
spectrum of the PBMC sample and, based on the second derivative of
the infrared (IR) spectrum, to generate an output indicative of the
presence of a solid tumor.
34. The system according to claim 33, wherein the IR spectrum
includes a Fourier Transformed Infrared (FTIR) spectrum, and
wherein the data processor is configured to calculate a second
derivative of the FTIR spectrum.
35. The system according to claim 32, wherein the data processor is
configured to analyze the infrared (IR) spectrum by assessing a
characteristic of the PBMC sample at at least one wavenumber
selected from the group consisting of: 765.+-.4 cm-1, 798.+-.4
cm-1, 809.+-.4 cm-1, 814.+-.4 cm-1, 875.+-.4 cm-1, 997.+-.4 cm-1,
1001.+-.4 cm-1, 1015.+-.4 cm-1, 1103.+-.4 cm-1, 1118.+-.4 cm-1,
1162.+-.4 cm-1, 1221.+-.4 cm-1, 1270.+-.4 cm-1, 1283.+-.4 cm-1,
1295.+-.4 cm-1, 1315.+-.4 cm-1, 1341.+-.4 cm-1, 1367.+-.4 cm-1,
1392.+-.4 cm-1, 1429.+-.4 cm-1, 1440.+-.4 cm-1, 1445.+-.4 cm-1, and
1455.+-.4 cm-1.
36. The system according to claim 35, wherein the data processor is
configured to analyze the infrared (IR) spectrum by assessing the
characteristic at at least two wavenumbers selected from the
group.
37. The system according to claim 35, wherein the data processor is
configured to analyze the infrared (IR) spectrum by assessing the
characteristic at at least three wavenumbers selected from the
group.
38. A computer program product for administering processing of a
body of data, the product comprising a computer-readable medium
having program instructions embodied therein, which instructions,
when read by a computer, cause the computer to: obtain an infrared
(IR) spectrum of a Peripheral Blood Mononuclear Cells (PBMC) by
analyzing the PBMC by infrared spectroscopy; and based on the
infrared spectrum, generate an output indicative of the presence of
a solid tumor.
39. A method comprising: obtaining an infrared (IR) spectrum of a
Peripheral Blood Mononuclear Cells (PBMC) sample by analyzing the
sample by infrared spectroscopy; and based on the infrared
spectrum, generating an output indicative of the presence of a
solid tumor in a breast tissue of a subject.
40. The method according to claim 39, wherein analyzing comprises
assessing a characteristic of the sample at at least one wavenumber
selected from the group consisting of: 752.+-.4 cm-1, 1030.+-.4
cm-1, 1046.+-.4 cm-1, 1128.+-.4 cm-1, and 1237.+-.4 cm-1.
41. The method according to claim 40, wherein analyzing comprises
assessing the characteristic at at least two wavenumbers selected
from the group.
42. The method according to claim 40, wherein analyzing comprises
assessing the characteristic at at least three wavenumbers selected
from the group.
43. The method according to claim 39, wherein analyzing the sample
by infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared (FTIR) spectroscopy, and wherein
obtaining the infrared (IR) spectrum comprises obtaining a Fourier
Transformed Infrared (FTIR) spectrum.
44. The method according to claim 43, wherein analyzing the sample
by infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared microspectroscopy (FTIR-MSP).
45. A method comprising: obtaining an infrared (IR) spectrum of a
Peripheral Blood Mononuclear Cells (PBMC) sample by analyzing the
sample by infrared spectroscopy; and based on the infrared
spectrum, generating an output indicative of the presence of a
solid tumor in tissue of a gastrointestinal tract of a subject.
46. The method according to claim 45, wherein analyzing comprises
assessing a characteristic of the sample at at least one wavenumber
selected from the group consisting of: 797.+-.4 cm-1, 830.+-.4
cm-1, 893.+-.4 cm-1, 899.+-.4 cm-1, 1128.+-.4 cm-1, 1298.+-.4 cm-1,
1354.+-.4 cm-1, 1714.+-.4 cm-1 1725.+-.4 cm-1, 1738,.+-.4 cm-1, and
3013.+-.4 cm-1.
47. The method according to claim 46, wherein analyzing comprises
assessing the characteristic at at least two wavenumbers selected
from the group.
48. The method according to claim 46, wherein analyzing comprises
assessing the characteristic at at least three wavenumbers selected
from the group.
49. The method according to claim 45, wherein analyzing the sample
by infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared (FTIR) spectroscopy, and wherein
obtaining the infrared (IR) spectrum comprises obtaining a Fourier
Transformed Infrared (FTIR) spectrum.
50. The method according to claim 49, wherein analyzing the sample
by infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared microspectroscopy (FTIR-MSP).
51. A method comprising: obtaining an infrared (IR) spectrum of a
Peripheral Blood Mononuclear Cells (PBMC) sample by analyzing the
sample by infrared spectroscopy; and based on the infrared
spectrum, generating an output indicative of the presence of a
solid tumor in lung tissue of a subject.
52. The method according to claim 51, wherein analyzing comprises
assessing a characteristic of the sample at at least one wavenumber
selected from the group consisting of: 765.+-.4 cm-1, 780.+-.4
cm-1, 797.+-.4 cm-1, 851.+-.4 cm-1, 874.+-.4 cm-1, 881.+-.4 cm-1,
913.+-.4 cm-1, 923.+-.4 cm-1, 958.+-.4 cm-1, 968,.+-.4 cm-1,
1044.+-.4 cm-1, 1085.+-.4 cm-1, 1191.+-.4 cm-1, 1241.+-.4 cm-1,
1344.+-.4 cm-1, 1373.+-.4 cm-1, 1417.+-.4 cm-1, 1458.+-.4 cm-1,
1469.+-.4 cm-1, 1692.+-.4 cm-1, 1714.+-.4 cm-1, 1728.+-.4 cm-1,
2852.+-.4 cm-1, and 2984.+-.4 cm.
53. The method according to claim 52, wherein analyzing comprises
assessing the characteristic at at least two wavenumbers selected
from the group.
54. The method according to claim 52, wherein analyzing comprises
assessing the characteristic at at least three wavenumbers selected
from the group.
55. The method according to claim 51, wherein analyzing the sample
by infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared (FTIR) spectroscopy, and wherein
obtaining the infrared (IR) spectrum comprises obtaining a Fourier
Transformed Infrared (FTIR) spectrum.
56. The method according to claim 55, wherein analyzing the sample
by infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared microspectroscopy (FTIR-MSP).
57. A method comprising: obtaining an infrared (IR) spectrum of a
Peripheral Blood Mononuclear Cells (PBMC) sample by analyzing the
sample by infrared spectroscopy; and based on the infrared
spectrum, generating an output indicative of the presence of a
solid tumor in a prostate tissue of a subject.
58. The method according to claim 57, wherein analyzing comprises
assessing a characteristic of the sample at at least one wavenumber
selected from the group consisting of: 828.+-.4 cm-1, 932.+-.4
cm-1, 997.+-.4 cm-1, 1059.+-.4 cm-1, 1299.+-.4 cm-1, 1302.+-.4
cm-1, 1403.+-.4 cm-1, 1454.+-.4 cm-1, 1714.+-.4 cm-1, 2979,.+-.4
cm-1, and 3013.+-.4 cm-1.
59. The method according to claim 58, wherein analyzing comprises
assessing the characteristic at at least two wavenumbers selected
from the group.
60. The method according to claim 58, wherein analyzing comprises
assessing the characteristic at at least three wavenumbers selected
from the group.
61. The method according to claim 57, wherein analyzing the sample
by infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared (FTIR) spectroscopy, and wherein
obtaining the infrared (IR) spectrum comprises obtaining a Fourier
Transformed Infrared (FTIR) spectrum.
62. The method according to claim 61, wherein analyzing the sample
by infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared microspectroscopy (FTIR-MSP).
63. A method comprising: obtaining an infrared (IR) spectrum of a
Peripheral Blood Mononuclear Cells (PBMC) sample from a cancer
patient by analyzing the sample by infrared spectroscopy; and based
on the infrared spectrum, generating an output indicative of a
stage of the cancer.
64. The method according to claim 63, wherein analyzing comprises
assessing a characteristic of the sample at at least one wavenumber
selected from the group consisting of: 865.+-.4 cm-1, 897.+-.4
cm-1, 924.+-.4 cm-1, 1030.+-.4 cm-1, 1047.+-.4 cm-1, 1191.+-.4
cm-1, and 1238.+-.4 cm-1.
65. The method according to claim 64, wherein analyzing comprises
assessing the characteristic at at least two wavenumbers selected
from the group.
66. The method according to claim 64, wherein analyzing comprises
assessing the characteristic at at least three wavenumbers selected
from the group.
67. The method according to claim 63, wherein analyzing the sample
by infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared (FTIR) spectroscopy, and wherein
obtaining the infrared (IR) spectrum comprises obtaining a Fourier
Transformed Infrared (FTIR) spectrum.
68. The method according to claim 67, wherein analyzing the sample
by infrared (IR) spectroscopy comprises analyzing the sample by
Fourier Transformed Infrared microspectroscopy (FTIR-MSP).
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] The present application claims the priority of U.S.
Provisional Application 61/350,073 to Kapelushnik et al., filed
Jun. 1, 2010, which is incorporated herein by reference.
FIELD OF EMBODIMENTS OF THE INVENTION
[0002] Embodiments of the present invention relate generally to
diagnosis and monitoring of cancer, and particularly to methods for
diagnosis and monitoring of malignant solid tumors.
BACKGROUND
[0003] Infrared spectroscopy is a technique based on the absorption
or reflection of infrared radiation by chemical substances; each
chemical substance having unique absorption spectra. Fourier
Transform Infrared (FTIR) spectroscopy is used to identify
biochemical compounds and examine the biochemical composition of a
biological sample. Typically, FTIR spectra are composed of
absorption bands each corresponding to specific functional groups
related to cellular components such as lipids, proteins,
carbohydrates and nucleic acids. Processes such as carcinogenesis
may trigger global changes in cancer cell biochemistry resulting in
differences in the absorption spectra when analyzed by FTIR
spectroscopy techniques. Therefore, FTIR spectroscopy is commonly
used to distinguish between normal and abnormal tissue by analyzing
the changes in absorption bands of macromolecules such as lipids,
proteins, carbohydrates and nucleic acids. Additionally, FTIR
spectroscopy may be utilized for evaluation of cell death mode,
cell cycle progression and the degree of maturation of
hematopoietic cells.
SUMMARY OF EMBODIMENTS OF THE INVENTION
[0004] In some applications of the present invention, methods and
systems are provided for the diagnosis and monitoring of multiple
types of malignant neoplasms, particularly malignant solid
tumors.
[0005] Additionally or alternatively, some applications of the
present invention comprise diagnosis and monitoring of a
pre-malignant condition.
[0006] Typically "Total Biochemical Infrared Analysis" (TBIA) of
blood-derived mononuclear cells is used to diagnose a solid tumor.
For example, some applications of the present invention comprise
analysis by infrared (IR) spectroscopy, e.g., FTIR spectroscopy and
microspectroscopy, of global biochemical properties of
blood-derived mononuclear cells for the detection of solid tumors.
As provided by some applications of the present invention, FTIR
Optical Diagnosis Technology (FODT) analysis of biochemical changes
in a Peripheral Blood Mononuclear Cells (PBMC) sample of a patient
can indicate the presence of a solid tumor and/or a pre-malignant
condition.
[0007] For some applications, biochemical analysis of PBMC obtained
from cancer patients and from control individuals who do not suffer
from a malignant solid tumor, e.g., healthy controls, is conducted
using FTIR microspectroscopy techniques. In accordance with some
applications of the present invention, PBMC from a plurality of
cancer patients each suffering from a solid tumor (e.g., in the
breast, pancreas, lung, head and neck, prostate, ovary, and
gastrointestinal tract) is analyzed by FTIR microspectroscopy
techniques. Subsequently, the FTIR spectra (absorption and/or
reflection) of the PBMC samples of the cancer patients are compared
to the FTIR spectra of PBMC samples obtained from the controls.
[0008] The inventors have identified that the PBMC samples obtained
from cancer patients suffering from a malignant solid tumor produce
FTIR spectra that differ from those of the control individuals who
do not suffer from a malignant solid tumor, allowing distinguishing
between the cancer patients and controls. Thus, some applications
of the present invention can be used to diagnose cancer patients
suffering from various types of malignancies, particularly solid
tumors. Importantly, the distinction by FTIR spectroscopy between
controls and patients suffering from solid tumors is typically
performed based on analysis of PBMC and not of the actual tumor
cells.
[0009] For some applications, a data processor analyzes the IR
spectrum, e.g., the FTIR spectrum, of the PBMC sample of a subject.
Information from the data processor is typically fed into an output
unit that generates a result indicative of the presence of a solid
tumor and/or a pre-malignant condition, based on the infrared (IR)
spectrum. Additionally, the data processor is typically configured
to calculate a second derivative of the infrared (IR) spectrum of
the PBMC sample and, based on the second derivative of the infrared
(IR) spectrum, to generate an output indicative of the presence of
a solid tumor.
[0010] Additionally, the inventors have identified that PBMC
obtained from each cancer patient suffering from a solid tumor
produced an FTIR spectrum having a unique spectral pattern which is
characteristic of the type of malignancy, e.g., breast, lung,
prostate or gastrointestinal, and distinct from spectra of other
malignancy types.
[0011] For some applications, analysis by IR spectroscopy, e.g.,
FTIR spectroscopy, of the biochemistry of PBMC or any other
blood-derived cells is used for the screening of large populations,
aiding in the early detection of solid tumors. FTIR spectroscopy
(and microspectroscopy) is typically a simple, reagent-free and
rapid method suitable for use as a screening test for large
populations. Early detection of cancer generally enables early
intervention and treatment, contributing to a reduced mortality
rate.
[0012] There is therefore provided in accordance with some
applications of the present invention a method including:
[0013] obtaining an infrared (IR) spectrum of a Peripheral Blood
Mononuclear Cells (PBMC) sample by analyzing the sample by infrared
spectroscopy; and
[0014] based on the infrared spectrum, generating an output
indicative of the presence of a solid tumor or a pre-malignant
condition.
[0015] For some applications, generating the output includes
generating the output indicative of the presence of the solid
tumor.
[0016] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared (FTIR) spectroscopy, and obtaining the infrared (IR)
spectrum includes obtaining a Fourier Transformed Infrared (FTIR)
spectrum.
[0017] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared microspectroscopy (FTIR-MSP).
[0018] For some applications, analyzing includes assessing a
characteristic of the sample at at least one wavenumber selected
from the group consisting of: 765.+-.4 cm-1, 798.+-.4 cm-1,
809.+-.4 cm-1, 814.+-.4 cm-1, 875.+-.4 cm-1, 997.+-.4 cm-1,
1001.+-.4 cm-1, 1015.+-.4 cm-1, 1103.+-.4 cm-1, 1118.+-.4 cm-1,
1162.+-.4 cm-1, 1221.+-.4 cm-1, 1270.+-.4 cm-1, 1283.+-.4 cm-1,
1295.+-.4 cm-1, 1315.+-.4 cm-1, 1341.+-.4 cm-1, 1367.+-.4 cm-1,
1392.+-.4 cm-1, 1429.+-.4 cm-1, 1440.+-.4 cm-1, 1445.+-.4 cm-1 and
1455.+-.4 cm-1.
[0019] For some applications, analyzing includes assessing the
characteristic at at least two wavenumbers selected from the
group.
[0020] For some applications, analyzing includes assessing the
characteristic at at least three wavenumbers selected from the
group.
[0021] For some applications, assessing the characteristic includes
analyzing a band of the IR spectrum surrounding at least one
wavenumber selected from the group.
[0022] For some applications, analyzing the sample includes
obtaining a second derivative of the infrared (IR) spectrum of the
sample.
[0023] For some applications, the infrared (IR) spectrum includes
an absorption spectrum, and obtaining the infrared (IR) spectrum
includes obtaining the absorption spectrum.
[0024] For some applications, the infrared (IR) spectrum includes a
reflection spectrum, and obtaining the infrared (IR) spectrum
includes obtaining the reflection spectrum.
[0025] For some applications, generating the output includes
indicating via the output whether the solid tumor is a first type
of solid tumor or a second type of solid tumor.
[0026] For some applications, the solid tumor includes a solid
tumor in tissue selected from the group consisting of: head and
neck, esophagus, and pancreas, and generating the output includes
generating an output indicative of the presence of a solid tumor in
tissue selected from the group.
[0027] For some applications, the solid tumor includes a solid
tumor in tissue selected from the group consisting of: breast,
gastrointestinal tract, prostate, and lung, and generating the
output includes generating an output indicative of the presence of
a solid tumor in tissue selected from the group.
[0028] For some applications, analyzing includes assessing a
characteristic of the sample at at least one wavenumber selected
from the group consisting of: 752.+-.4 cm-1, 1030.+-.4 cm-1,
1046.+-.4 cm-1, 1128.+-.4 cm-1, and 1237.+-.4 cm-1, and generating
includes generating an output indicative of the presence of a tumor
in the breast tissue.
[0029] For some applications, analyzing includes assessing a
characteristic of the sample at at least one wavenumber selected
from the group consisting of: 797.+-.4 cm-1, 830.+-.4 cm-1,
893.+-.4 cm-1, 899.+-.4 cm-1, 1128.+-.4 cm-1, 1298.+-.4 cm-1,
1354.+-.4 cm-1, 1714.+-.4 cm-1 1725.+-.4 cm-1, 1738,.+-.4 cm-1, and
3013.+-.4 cm-1, and generating includes generating an output
indicative of the presence of a tumor in the gastrointestinal tract
tissue.
[0030] For some applications, analyzing includes assessing a
characteristic of the sample at at least one wavenumber selected
from the group consisting of: 765.+-.4 cm-1, 780.+-.4 cm-1,
797.+-.4 cm-1, 851.+-.4 cm-1, 874.+-.4 cm-1, 881.+-.4 cm-1,
913.+-.4 cm-1, 923.+-.4 cm-1, 958.+-.4 cm-1, 968,.+-.4 cm-1,
1044.+-.4 cm-1, 1085.+-.4 cm-1, 1191.+-.4 cm-1, 1241.+-.4 cm-1,
1344.+-.4 cm-1, 1373.+-.4 cm-1, 1417.+-.4 cm-1, 1458.+-.4 cm-1,
1469.+-.4 cm-1, 1692.+-.4 cm-1, 1714.+-.4 cm-1, 1728.+-.4 cm-1,
2852.+-.4 cm-1, and 2984.+-.4 cm, and generating includes
generating an output indicative of the presence of a tumor in the
lung tissue.
[0031] For some applications, analyzing includes assessing a
characteristic of the sample at at least one wavenumber selected
from the group consisting of: 828.+-.4 cm-1, 932.+-.4 cm-1,
997.+-.4 cm-1, 1059.+-.4 cm-1, 1299.+-.4 cm-1, 1302.+-.4 cm-1,
1403.+-.4 cm-1, 1454.+-.4 cm-1, 1714.+-.4 cm-1, 2979,.+-.4 cm-1,
and 3013.+-.4 cm-1, and generating includes generating an output
indicative of the presence of a tumor in the prostate tissue.
[0032] There is further provided in accordance with some
applications of the present invention a method including:
[0033] obtaining an infrared (IR) spectrum of a sample of white
blood cells by analyzing the sample by infrared spectroscopy;
and
[0034] based on the infrared spectrum, generating an output
indicative of the presence of a solid tumor or a pre-malignant
condition.
[0035] For some applications, generating the output includes
generating the output indicative of the presence of the solid
tumor.
[0036] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared (FTIR) spectroscopy, and obtaining the infrared (IR)
spectrum includes obtaining a Fourier Transformed Infrared (FTIR)
spectrum.
[0037] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared microspectroscopy (FTIR-MSP).
[0038] There is additionally provided in accordance with some
applications of the present invention a method for monitoring the
effect of an anti-cancer treatment on a subject undergoing
anti-cancer treatment for a solid tumor, for use with a first
Peripheral Blood Mononuclear Cells (PBMC) sample separated from
blood of the subject that was obtained prior to initiation of the
treatment and a second PBMC sample separated from blood of the
subject that was obtained after initiation of the treatment, the
method including:
[0039] obtaining IR spectra of the first and second PBMC samples by
analyzing the first and second PBMC samples by IR spectroscopy;
and
[0040] based on the IR spectra, generating an output indicative of
the effect of the treatment.
[0041] For some applications, analyzing the first and second PBMC
samples by IR spectroscopy includes analyzing the samples by
Fourier Transformed Infrared spectroscopy, and obtaining the IR
spectra includes obtaining Fourier Transformed Infrared (FTIR)
spectra.
[0042] For some applications, analyzing the first and second PBMC
samples by infrared (IR) spectroscopy includes analyzing the first
and second PBMC samples by Fourier Transformed Infrared
microspectroscopy (FTIR-MSP).
[0043] For some applications, the method includes obtaining an IR
spectrum of a third PBMC sample separated from blood of the subject
that was obtained following termination of the treatment, by
analyzing the third PBMC sample by IR spectroscopy.
[0044] For some applications, analyzing includes assessing a
characteristic of the sample at at least one wavenumber selected
from the group consisting of: 765.+-.4 cm-1, 798.+-.4 cm-1,
809.+-.4 cm-1, 814.+-.4 cm-1, 875.+-.4 cm-1, 997.+-.4 cm-1,
1001.+-.4 cm-1, 1015.+-.4 cm-1, 1103.+-.4 cm-1, 1118.+-.4 cm-1,
1162.+-.4 cm-1, 1221.+-.4 cm-1, 1270.+-.4 cm-1, 1283.+-.4 cm-1,
1295.+-.4 cm-1, 1315.+-.4 cm-1, 1341.+-.4 cm-1, 1367.+-.4 cm-1,
1392.+-.4 cm-1, 1429.+-.4 cm-1, 1440.+-.4 cm-1, 1445.+-.4 cm-1, and
1455.+-.4 cm-1.
[0045] For some applications, analyzing includes assessing the
characteristic at at least two wavenumbers selected from the
group.
[0046] For some applications, analyzing includes assessing the
characteristic at at least three wavenumbers selected from the
group.
[0047] There is yet additionally provided in accordance with some
applications of the present invention, a method including:
[0048] obtaining an infrared (IR) spectrum of a Peripheral Blood
Mononuclear Cells (PBMC) sample by analyzing the sample; and
[0049] based on the infrared spectrum, generating an output
indicative of the presence of a solid tumor or a pre-malignant
condition.
[0050] For some applications, generating the output includes
generating the output indicative of the presence of the solid
tumor.
[0051] There is still additionally provided in accordance with some
applications of the present invention, a system for diagnosing a
solid tumor, including:
[0052] a data processor, configured to analyze an infrared (IR)
spectrum of a Peripheral Blood Mononuclear Cells (PBMC) sample of a
subject; and
[0053] an output unit, configured to generate an output indicative
of the presence of a solid tumor, based on the infrared (IR)
spectrum.
[0054] For some applications, the data processor is configured to
calculate a second derivative of the infrared (IR) spectrum of the
PBMC sample and, based on the second derivative of the infrared
(IR) spectrum, to generate an output indicative of the presence of
a solid tumor.
[0055] For some applications, the IR spectrum includes a Fourier
Transformed Infrared (FTIR) spectrum, and the data processor is
configured to calculate a second derivative of the FTIR
spectrum.
[0056] For some applications, the data processor is configured to
analyze the infrared (IR) spectrum by assessing a characteristic of
the PBMC sample at at least one wavenumber selected from the group
consisting of: 765.+-.4 cm-1, 798.+-.4 cm-1, 809.+-.4 cm-1,
814.+-.4 cm-1, 875.+-.4 cm-1, 997.+-.4 cm-1, 1001.+-.4 cm-1,
1015.+-.4 cm-1, 1103.+-.4 cm-1, 1118.+-.4 cm-1, 1162.+-.4 cm-1,
1221.+-.4 cm-1, 1270.+-.4 cm-1, 1283.+-.4 cm-1, 1295.+-.4 cm-1,
1315.+-.4 cm-1, 1341.+-.4 cm-1, 1367.+-.4 cm-1, 1392.+-.4 cm-1,
1429.+-.4 cm-1, 1440.+-.4 cm-1, 1445.+-.4 cm-1, and 1455.+-.4
cm-1.
[0057] For some applications, the data processor is configured to
analyze the infrared (IR) spectrum by assessing the characteristic
at at least two wavenumbers selected from the group.
[0058] For some applications, the data processor is configured to
analyze the infrared (IR) spectrum by assessing the characteristic
at at least three wavenumbers selected from the group.
[0059] There is still further provided in accordance with some
applications of the present invention, a computer program product
for administering processing of a body of data, the product
including a computer-readable medium having program instructions
embodied therein, which instructions, when read by a computer,
cause the computer to:
[0060] obtain an infrared (IR) spectrum of a Peripheral Blood
Mononuclear Cells (PBMC) by analyzing the PBMC by infrared
spectroscopy; and
[0061] based on the infrared spectrum, generate an output
indicative of the presence of a solid tumor.
[0062] There is yet provided in accordance with some applications
of the present invention, a method including:
[0063] obtaining an infrared (IR) spectrum of a Peripheral Blood
Mononuclear Cells (PBMC) sample by analyzing the sample by infrared
spectroscopy; and
[0064] based on the infrared spectrum, generating an output
indicative of the presence of a solid tumor in a breast tissue of a
subject.
[0065] For some applications, analyzing includes assessing a
characteristic of the sample at at least one wavenumber selected
from the group consisting of: 752.+-.4 cm-1, 1030.+-.4 cm-1,
1046.+-.4 cm-1, 1128.+-.4 cm-1, and 1237.+-.4 cm-1.
[0066] For some applications, analyzing includes assessing the
characteristic at at least two wavenumbers selected from the
group.
[0067] For some applications, analyzing includes assessing the
characteristic at at least three wavenumbers selected from the
group.
[0068] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared (FTIR) spectroscopy, and obtaining the infrared (IR)
spectrum includes obtaining a Fourier Transformed Infrared (FTIR)
spectrum.
[0069] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared microspectroscopy (FTIR-MSP).
[0070] There is still provided in accordance with some applications
of the present invention a method including:
[0071] obtaining an infrared (IR) spectrum of a Peripheral Blood
Mononuclear Cells (PBMC) sample by analyzing the sample by infrared
spectroscopy; and
[0072] based on the infrared spectrum, generating an output
indicative of the presence of a solid tumor in tissue of a
gastrointestinal tract of a subject.
[0073] For some applications, analyzing includes assessing a
characteristic of the sample at at least one wavenumber selected
from the group consisting of: 797.+-.4 cm-1, 830.+-.4 cm-1,
893.+-.4 cm-1, 899.+-.4 cm-1, 1128.+-.4 cm-1, 1298.+-.4 cm-1,
1354.+-.4 cm-1, 1714.+-.4 cm-1 1725.+-.4 cm-1, 1738,.+-.4 cm-1, and
3013.+-.4 cm-1.
[0074] For some applications, analyzing includes assessing the
characteristic at at least two wavenumbers selected from the
group.
[0075] For some applications, analyzing includes assessing the
characteristic at at least three wavenumbers selected from the
group.
[0076] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared (FTIR) spectroscopy, and obtaining the infrared (IR)
spectrum includes obtaining a Fourier Transformed Infrared (FTIR)
spectrum.
[0077] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared microspectroscopy (FTIR-MSP).
[0078] There is additionally provided in accordance with some
applications of the present invention, a method including:
[0079] obtaining an infrared (IR) spectrum of a Peripheral Blood
Mononuclear Cells (PBMC) sample by analyzing the sample by infrared
spectroscopy; and
[0080] based on the infrared spectrum, generating an output
indicative of the presence of a solid tumor in lung tissue of a
subject.
[0081] For some applications, analyzing includes assessing a
characteristic of the sample at at least one wavenumber selected
from the group consisting of: 765.+-.4 cm-1, 780.+-.4 cm-1,
797.+-.4 cm-1, 851.+-.4 cm-1, 874.+-.4 cm-1, 881.+-.4 cm-1,
913.+-.4 cm-1, 923.+-.4 cm-1, 958.+-.4 cm-1, 968,.+-.4 cm-1,
1044.+-.4 cm-1, 1085.+-.4 cm-1, 1191.+-.4 cm-1, 1241.+-.4 cm-1,
1344.+-.4 cm-1, 1373.+-.4 cm-1, 1417.+-.4 cm-1, 1458.+-.4 cm-1,
1469.+-.4 cm-1, 1692.+-.4 cm-1, 1714.+-.4 cm-1, 1728.+-.4 cm-1,
2852.+-.4 cm-1, and 2984.+-.4 cm.
[0082] For some applications, analyzing includes assessing the
characteristic at at least two wavenumbers selected from the
group.
[0083] For some applications, analyzing includes assessing the
characteristic at at least three wavenumbers selected from the
group.
[0084] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared (FTIR) spectroscopy, and obtaining the infrared (IR)
spectrum includes obtaining a Fourier Transformed Infrared (FTIR)
spectrum.
[0085] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared microspectroscopy (FTIR-MSP).
[0086] There is further additionally provided in accordance with
some applications of the present invention, a method including:
[0087] obtaining an infrared (IR) spectrum of a Peripheral Blood
Mononuclear Cells (PBMC) sample by analyzing the sample by infrared
spectroscopy; and
[0088] based on the infrared spectrum, generating an output
indicative of the presence of a solid tumor in a prostate tissue of
a subject.
[0089] For some applications, analyzing includes assessing a
characteristic of the sample at at least one wavenumber selected
from the group consisting of: 828.+-.4 cm-1, 932.+-.4 cm-1,
997.+-.4 cm-1, 1059.+-.4 cm-1, 1299.+-.4 cm-1, 1302.+-.4 cm-1,
1403.+-.4 cm-1, 1454.+-.4 cm-1, 1714.+-.4 cm-1, 2979,.+-.4 cm-1,
and 3013.+-.4 cm-1.
[0090] For some applications, analyzing includes assessing the
characteristic at at least two wavenumbers selected from the
group.
[0091] For some applications, analyzing includes assessing the
characteristic at at least three wavenumbers selected from the
group.
[0092] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared (FTIR) spectroscopy, and obtaining the infrared (IR)
spectrum includes obtaining a Fourier Transformed Infrared (FTIR)
spectrum.
[0093] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared microspectroscopy (FTIR-MSP).
[0094] There is further provided in accordance with some
applications of the present invention, a method including:
[0095] obtaining an infrared (IR) spectrum of a Peripheral Blood
Mononuclear Cells (PBMC) sample from a cancer patient by analyzing
the sample by infrared spectroscopy; and
[0096] based on the infrared spectrum, generating an output
indicative of a stage of the cancer.
[0097] For some applications, analyzing includes assessing a
characteristic of the sample at at least one wavenumber selected
from the group consisting of: 865.+-.4 cm-1, 897.+-.4 cm-1,
924.+-.4 cm-1, 1030.+-.4 cm-1, 1047.+-.4 cm-1, 1191.+-.4 cm-1, and
1238.+-.4 cm-1.
[0098] For some applications, analyzing includes assessing the
characteristic at at least two wavenumbers selected from the
group.
[0099] For some applications, analyzing includes assessing the
characteristic at at least three wavenumbers selected from the
group.
[0100] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared (FTIR) spectroscopy, and obtaining the infrared (IR)
spectrum includes obtaining a Fourier Transformed Infrared (FTIR)
spectrum.
[0101] For some applications, analyzing the sample by infrared (IR)
spectroscopy includes analyzing the sample by Fourier Transformed
Infrared microspectroscopy (FTIR-MSP).
[0102] The present invention will be more fully understood from the
following detailed description of embodiments thereof, taken
together with the drawings, in which:
BRIEF DESCRIPTION OF THE DRAWINGS
[0103] FIGS. 1A-C are graphs representing FTIR absorption spectra
and the second derivative of absorption spectra and analysis
thereof, based on PBMC samples from several cancer patients and
controls, derived in accordance with some applications of the
present invention;
[0104] FIGS. 2A-C are graphs representing FTIR absorption spectra
and the second derivative of absorption spectra of PBMC from a
pancreatic cancer patient compared to PBMC from healthy controls,
derived in accordance with some applications of the present
invention;
[0105] FIGS. 3A-C are graphs showing the second derivative of
spectra of PBMC from several cancer patients and healthy controls,
derived in accordance with some applications of the present
invention, and a table summarizing the main biochemical changes
induced in PBMC of the cancer patients, as observed by FTIR
microspectroscopy, as derived in accordance with some applications
of the present invention; and
[0106] FIG. 4 shows second derivative spectra of PBMC from a breast
cancer patient, a gastrointestinal cancer patient with a history of
breast cancer, and healthy controls, as derived in accordance with
some applications of the present invention; and
[0107] FIGS. 5A-G are graphs representing the second derivative
spectra of PBMC and analysis thereof, based on PBMC samples from
cancer patients suffering from various types of solid tumors,
derived in accordance with some applications of the present
invention; and
[0108] FIGS. 6A-C are graphs representing the second derivative
spectra and analysis thereof, of PBMC samples from cancer patients
suffering from different stages of cancer, derived in accordance
with some applications of the present invention.
DETAILED DESCRIPTION OF THE EMBODIMENTS
[0109] Some applications of the present invention comprise
diagnosis of a solid tumor by techniques of IR spectroscopy, e.g.,
FTIR microspectroscopy (MSP) techniques. For some applications,
FTIR Optical Diagnosis Technology (FODT) is used to diagnose a
solid tumor based on biochemical proprieties of Peripheral Blood
Mononuclear Cells (PBMC) of a subject. Some applications of the
present invention comprise obtaining a blood sample from a subject
and analyzing mononuclear cells from the sample by FTIR-MSP
techniques for the detection of a solid tumor. Typically, a PBMC
sample of a patient suffering from a solid tumor is identified as
exhibiting FTIR spectra that are different from FTIR spectra
produced by PBMC from a subject who does not suffer from a solid
tumor (for some applications, the control group may include
subjects suffering from a pathology that is not a solid tumor).
Accordingly, some applications of the present invention provide a
useful method for the detection of cancer, specifically solid
tumors. FTIR spectra of PBMC obtained from a cancer patient with a
solid tumor generally reflect biochemical changes which occur in
the PBMC of the patient in response to the tumor.
[0110] For some applications, methods of the present invention are
used to determine a stage of the cancer.
[0111] For some applications, methods of the present invention can
be used to provide monitoring and follow up of cancer patients
during and after treatment, e.g., chemotherapy treatment.
Typically, changes in FTIR spectra of PBMC of solid-tumor patients
who are undergoing treatment can indicate biochemical changes in
the cells in response to the treatment. This biochemical
information can provide insight into the effect of treatment on the
patient and/or the tumor.
[0112] In some applications of the present invention, analysis of
PBMC by FTIR-MSP is used to detect a type of solid tumor.
Typically, each type of malignant solid tumor produces distinct
FTIR spectra of the PBMC, which are unique to the type of solid
tumor. This can be due to each type of solid tumor inducing
specific biochemical changes in PBMC.
Methods Used in Some Embodiments of the Present Invention
[0113] A series of protocols are described hereinbelow which may be
used separately or in combination, as appropriate, in accordance
with applications of the present invention. It is to be appreciated
that numerical values are provided by way of illustration and not
limitation. Typically, but not necessarily, each value shown is an
example selected from a range of values that is within 20% of the
value shown. Similarly, although certain steps are described with a
high level of specificity, a person of ordinary skill in the art
will appreciate that other steps may be performed, mutatis
mutandis.
[0114] In accordance with some applications of the present
invention, the following methods were applied:
Obtaining Patient and Control Populations
[0115] All studies were approved by the local Ethics Committee of
the Soroka University Medical Center and conducted in accordance
with the Declaration of Helsinki. Qualified personnel obtained
informed consent from each individual participating in this
study.
[0116] The patient population included cancer patients (n=63)
diagnosed with primary solid tumors as set forth in the following
Table I:
TABLE-US-00001 TABLE I Disease site Control Breast GI Lung
Pro-state Other Gender Male 24 1 9 7 5 2 Female 22 24 9 1 0 5
Disease stage I 0 9 1 0 2 2 II 0 12 3 0 0 0 III 0 3 4 1 0 3 IV 0 1
10 7 3 2 Total 46 25 18 8 5 7
[0117] The patient population categorized under "other" included
six patients each diagnosed with a different type of primary tumor.
Among the "other" group are patients suffering from primary tumors
such as pancreas (n=2), head and neck (n=1) and esophagus
(n=1).
[0118] The diagnosis of cancer was determined by clinical,
surgical, histological, and pathologic diagnosis. The pathologic
stage of the tumor was determined according to
tumor-node-metastasis (TNM) classification, as described in "TNM
Classification of Malignant Tumours", by Sobin L H. et al., 7th
Edition, New York: John Wiley, 2009.
[0119] A control group (n=46) included healthy volunteers who
underwent detailed clinical questioning to exclude a possible
pathology, at the Soroka University Medical Center and Ben-Gurion
University.
Collection of Blood Samples
[0120] 1-2 ml of peripheral blood was collected in 5 ml EDTA blood
collection tubes (BD Vacutainer.RTM. Tubes, BD Vacutainer, Toronto)
from patients and healthy controls using standardized phlebotomy
procedures. Samples were processed within two hours of
collection.
[0121] Extraction of Peripheral Blood Mononuclear Cells (PBMC)
[0122] Platelet-depleted residual leukocytes obtained from cancer
patients and healthy controls were applied to Histopaque 1077
gradients (Sigma Chemical Co., St. Louis, Mo., USA) following
manufacturer's protocol to obtain PBMC.
[0123] The cells were aspirated from the interface, washed twice
with isotonic saline (0.9% NaCl solution) at 250 g, and resuspended
in 5 .mu.l fresh isotonic saline. 1.5 .mu.l of washed cells were
deposited on zinc selenide (ZnSe) slides to form approximately a
monolayer of cells. It is noted that any other suitable slide may
be used, e.g., reflection measurements may be carried out using a
gold slide. The slides were then air dried for 1 h under laminar
flow to remove water. The dried cells were than measured by FTIR
microscopy.
[0124] FTIR-Microspectroscopy
[0125] Fourier Transform Infrared Microspectroscopy (FTIR-MSP) and
Data Acquisition Measurements on cell cultures were performed using
the FTIR microscope IR scope 2 with a liquid-nitrogen-cooled
mercury-cadmium-telluride (MCT) detector, coupled to the FTIR
spectrometer Bruker Equinox model 55/S, using OPUS software (Bruker
Optik GmbH, Ettlingen, Germany). For some of the experiments,
Fourier Transform Infrared Microspectroscopy (FTIR-MSP) and Data
Acquisition Measurements were performed using the FTIR microscope
Nicolet Centaurus with a liquid-nitrogen-cooled
mercury-cadmium-telluride (MCT) detector, coupled to the FTIR
spectrometer Nicolet iS10, OMNIC software (Nicolet, Madison, Wis.)
using OPUS software (Bruker Optik GmbH, Ettlingen, Germany)
Essentially the same results were obtained with each of these
microscopes.
[0126] To achieve high signal-to-noise ratio (SNR), 128 coadded
scans were collected in each measurement in the wavenumber region
700 to 4000 cm-1. The measurement site was circular with a diameter
of 100 .mu.m and a spectral resolution of 4 cm-1 (0.482 cm-1 data
spacing). To reduce cell amount variation and facilitate proper
comparison between different samples, the following procedures were
adopted. [0127] 1. Each sample was measured at least five times at
different spots. [0128] 2. Analog to Digital Converter (ADC) rates
were empirically chosen between 2000 to 3000 counts/sec (providing
measurement areas with similar cellular density). [0129] 3. The
obtained spectra were baseline corrected using the rubber band
method, with 64 consecutive points, and normalized using vector
normalization in OPUS software as described in an article entitled
"Early spectral changes of cellular malignant transformation using
Fourier transformation infrared microspectroscopy," by Bogomolny et
al., 2007. J Biomed Opt.12:024003.
[0130] In order to obtain precise absorption values at a given
wavenumber with minimal background interference, the second
derivative spectra were used to determine concentrations of
bio-molecules of interest. This method is susceptible to changes in
full width at half maximum (FWHM) of the IR bands. However, in the
case of biological samples, all cells from the same type are
composed from similar basic components which give relatively broad
bands. Thus, it is possible to generally neglect the changes in
bands FWHM as described in an article entitled "Selenium alters the
lipid content and protein profile of rat heart: An FTIR
microspectroscopy study," by Toyran et al., Arch.Biochem.Biophys.
2007 458:184-193.
[0131] Statistical Analysis:
[0132] Statistical analysis was performed using STATISTICA software
(STATISTICA, StatSoft, Inc., Tulsa, Okla.) and the student T-test.
P-values <0.05 were considered significant.
Experimental Data
[0133] The experiments described hereinbelow were performed by the
inventors in accordance with applications of the present invention
and using the techniques described hereinabove.
[0134] The experiments presented hereinbelow with reference to
Example 1 and Example 2 demonstrate that in accordance with some
applications of the present invention, analysis of PBMC samples by
FTIR-MSP techniques can be used for diagnosis of a solid tumor
based on the FTIR-MSP spectral pattern at selected wavenumbers.
EXAMPLE 1
[0135] In a set of experiments, PBMC samples from 46 healthy
controls were analyzed by FTIR-MSP, and a typical FTIR-MSP spectral
pattern was established for control PBMC. Additionally, PBMC
samples from 63 cancer patients suffering from multiple types of
solid tumors were subjected to FTIR-MSP analysis and compared to
the control FTIR-MSP spectral pattern. The PBMC samples were
obtained by preliminary processing of the peripheral blood in
accordance with the protocols described hereinabove with reference
to extraction of peripheral blood mononuclear cells (PBMC). The
PBMC samples were then analyzed by FTIR-MSP, in accordance with the
protocols described hereinabove with reference to FTIR-MSP.
[0136] Reference is made to FIGS. 1A-C, which are graphs
representing FTIR absorption spectra and the second derivative of
the absorption spectra and analysis thereof, for PBMC samples from
63 cancer patients suffering from solid tumors and 46 healthy
controls, derived in accordance with some applications of the
present invention.
[0137] FTIR-MSP analysis of peripheral blood mononuclear cells
(PBMC) typically generated spectra in the region of 4000-700 cm-1.
The spectra are composed of several absorption bands, each
corresponding to specific functional groups of specific
macromolecules. FIG. 1A shows an average of the FTIR-MSP spectra of
PBMC samples of healthy controls and cancer patients in the regions
of 4000-700 cm-1, after baseline correction and vector
normalization. The spectra are composed of several absorption
bands, each corresponding to specific functional groups of specific
macromolecules such as lipids, proteins, and carbohydrates/nucleic
acids. The main absorption bands are marked. For example, the
region 3000-2830 cm-1 contains symmetric and antisymmetric
stretching of CH3 and CH2 groups, which correspond mainly to
proteins and lipids respectively. The region 1700-1500 cm-1
corresponds to amide I and amide II, which contain information
regarding the secondary structures of proteins. The region
1300-1000 cm-1 includes the symmetric and antisymmetric vibrations
of PO2-groups. 1000-700 cm-1 is a `finger print` region which
contains several different vibrations, corresponding to
carbohydrates, lipids, nucleic acids and other bio-molecules, as
described in an article by Mantsch M and Chapman D. entitled
"Infrared spectroscopy of bio molecules," John Wiley New York 1996.
The FTIR spectrum was typically analyzed by tracking changes in
absorption (intensity and/or shift) of these macromolecules.
[0138] Table II represents some of the main IR absorption bands for
PBMC cells, and their corresponding molecular functional
groups:
TABLE-US-00002 TABLE II Wavenumber (cm-1 .+-. 4) Assignment 2958
.nu..sub.as CH.sub.3, mostly proteins, lipids 2922 .nu..sub.as
CH.sub.2, mostly lipids, proteins 2873 .nu..sub.s CH.sub.3, mostly
proteins, lipids 2854 .nu..sub.s CH.sub.2, mostly lipids, proteins
~1,656 Amide I .nu. C.dbd.O (80%), .nu. C--N (10%), .delta. N--H
~1,546 Amide II .delta. N--H (60%), .nu. C--N (40%) 1400 .nu.
COO--, .delta. s CH3 lipids, proteins 1313 Amide III band
components of proteins 1240 .nu..sub.as PO.sub.2.sup.-,
phosphodiester groups of nucleic acids 1170 C--O bands from
glycomaterials and proteins 1155 .nu.C--O of proteins and
carbohydrates 1085 .nu.s PO2- of nucleic acids, phospholipids,
proteins 1053 .nu. C--O & .delta. C--O of carbohydrates 996
C--C & C--O of ribose of RNA 967 C--C & C--O of deoxyribose
skeletal motions of DNA 780 sugar-phosphate Z conformation of DNA
740 .nu. N.dbd.H of Thymine
[0139] Reference is made to FIG. 1B. In order to achieve effective
comparison between the PBMC samples of the cancer patients and the
controls, the second derivative of the baseline-corrected,
vector-normalized FTIR-MSP spectra was used. Results are presented
in FIG. 1B. As shown, the second derivative spectra of PBMC samples
from the cancer patients differed significantly from the second
derivative spectra of PBMC samples from the healthy controls in the
spectral region of 1340-1260 cm-1. The mean.+-.SEM is represented
by the hashed region (for control) and the dotted region (for
cancer) as shown in the exploded view in FIG. 1B.
[0140] Reference is made to FIG. 1C, which is a graph representing
values of the second derivative of absorption spectra at
wavenumbers A1-A23 of PBMC samples from cancer patients compared to
PBMC samples from healthy controls, derived in accordance with some
applications of the present invention. Statistical analysis was
performed and P-values are provided. As shown, the second
derivative of PBMC samples from the cancer patients differed
significantly from the second derivative analysis of FTIR-MSP
spectra from PBMC of healthy controls.
[0141] Table III lists the wavenumbers shown in FIG. 1C. Typically,
PBMC samples were analyzed by FTIR-MSP techniques using these
wavenumbers to distinguish between healthy controls and cancer
patients.
TABLE-US-00003 TABLE III Control vs. Cancer Wavenumber (cm-1 .+-.
4) A1 765 A2 798 A3 809 A4 814 A5 875 A6 997 A7 1001 A8 1015 A9
1103 A10 1118 A11 1162 A12 1221 A13 1270 A14 1283 A15 1295 A16 1315
A17 1341 A18 1367 A19 1392 A20 1429 A21 1440 A22 1445 A23 1455
EXAMPLE 2
[0142] In this set of experiments, PBMC from a single pancreatic
cancer patient was subjected to FTIR-MSP analysis and compared to a
control FTIR-MSP spectral pattern based on PBMC from 27 healthy
controls. Results are presented in FIGS. 2A-C. The PBMC was
obtained by preliminary processing of the peripheral blood in
accordance with the protocols described hereinabove with reference
to extraction of peripheral blood mononuclear cells (PBMC). The
PBMC samples were then analyzed by FTIR-MSP in accordance with the
protocols described hereinabove with reference to
FTIR-Microspectroscopy.
[0143] FIG. 2A shows representative FTIR-MSP spectra of PBMC of a
healthy control compared to FTIR-MSP spectra of PBMC of a
pancreatic cancer patient after baseline correction and Min-Max
normalization to amide II. Each spectrum represents the average of
five measurements at different sites for each sample. The spectra
are composed of several absorption bands, each corresponding to
specific functional groups of specific macromolecules such as
lipids, proteins, carbohydrates and nucleic acids. The main
absorption bands are marked. The FTIR spectrum was analyzed by
tracking changes in absorption (intensity and/or shift) of these
macromolecules.
[0144] As shown in FIG. 2A, the FTIR-MSP spectra derived from
analysis of PBMC from the pancreatic cancer patient exhibited a
different spectral pattern when compared to the FTIR-MSP spectra of
PBMC of healthy controls.
[0145] Reference is made to FIGS. 2B-C. In order to increase
accuracy and achieve effective comparison between the PBMC sample
of the pancreatic cancer patient and healthy controls, the second
derivative of the baseline-corrected, vector-normalized FTIR
spectra was used. Results are presented in FIGS. 2B-C. The main
absorption bands are marked. As shown, the second derivative
spectral pattern of PBMC from the pancreatic cancer patient
differed significantly from an average FTIR-MSP spectral pattern of
PBMC of the healthy controls. The mean.+-.SEM for the controls is
represented by the hashed region as shown in the exploded view in
FIGS. 2B-C.
[0146] Reference is now made to Example 3-Example 5. The
experiments presented hereinbelow with reference to Example
3-Example 5 demonstrate that in accordance with some applications
of the present invention, analysis of PBMC samples by FTIR-MSP
techniques is used to detect a type of solid tumor. Typically, each
type of malignant solid tumor produces distinct FTIR spectra of the
PBMC which are unique to the type of solid tumor.
EXAMPLE 3
[0147] In this set of preliminary experiments, PBMC from cancer
patients suffering from solid tumors, and PBMC from healthy
controls was analyzed by FTIR-MSP. The population of cancer
patients for this set of experiments comprised a total of 5
patients suffering from the following solid tumors: Breast (n=1),
lung (n=1), pancreas (n=1), head and neck (n=1) and esophagus
(n=1). The PBMC was obtained by preliminary processing of the
peripheral blood in accordance with the protocols described
hereinabove with reference to extraction of peripheral blood
mononuclear cells (PBMC). The PBMC samples were then analyzed by
FTIR-MSP in accordance with the protocols described hereinabove
with reference to FTIR-Microspectroscopy.
[0148] The results show that the FTIR-MSP spectral pattern of all
the cancer patients differs from those of the healthy controls. The
results additionally show that each type of malignancy produces a
distinct spectral absorption pattern of the PBMC, which is unique
to each type of solid tumor.
[0149] Reference is made to FIGS. 3A-B. In order to increase
accuracy and achieve effective comparison between PBMC samples of
cancer patients and healthy controls, the second derivative of the
baseline-corrected, vector-normalized FTIR spectra was used.
Results are presented in FIG. 3A. As shown in FIG. 3A, the second
derivative spectral pattern of PBMC from each one of the cancer
patients differed from PBMC of the healthy controls. For example,
the FTIR-MSP spectrum of PBMC of the lung cancer patient is
distinct from control by exhibiting decreased absorbance of
CH.sub.2, which corresponds to cellular lipids; a decrease in
v.sub.as PO.sub.2.sup.-; a shift to a higher wavenumber at 967
cm.sup.-1, which corresponds to deoxyribose skeletal motions of
DNA; and an increase in RNA absorption.
[0150] Additionally, each spectrum has its own unique spectral
pattern, which is distinct from control, and which is
characteristic of each type of malignancy.
[0151] Reference is made to FIG. 3B, which is a graph showing an
analysis of the second derivative data shown in FIG. 3A. FIG. 3B
represents the change in value of each type of cancer relative to
the control, as derived by analysis by FTIR-MSP of PBMC from each
patient. As shown, each cancer patient exhibited a spectrum that
differed from the control.
[0152] FIG. 3C is a table summarizing the main biochemical changes
induced in PBMC of cancer patients suffering from different types
of solid tumors, as observed by FTIR-MSP (shown in FIGS. 3A-B) and
analyzed in accordance with some applications of the present
invention. Peak intensities which indicate absorption were
calculated for each spectrum to reveal the main biochemical changes
characteristic of each type of tumor.
EXAMPLE 4
[0153] In this set of experiments, PBMC from a breast cancer
patient and a gastrointestinal cancer patient were analyzed by
FTIR-MSP, and compared to analysis of PBMC from healthy controls.
It is to be noted that the breast cancer patient suffers from a
primary breast tumor, and the gastrointestinal cancer patient
suffers from a primary gastrointestinal tumor. The gastrointestinal
cancer patient has a history of breast cancer, and a pathological
evaluation of the gastrointestinal tumor showed a breast cancer
phenotype rather than a gastrointestinal phenotype.
[0154] First, peripheral blood was extracted from the two cancer
patients and 27 healthy controls. The PBMC was obtained by
preliminary processing of the peripheral blood in accordance with
the protocols described hereinabove with reference to extraction of
peripheral blood mononuclear cells (PBMC). The PBMC samples were
then analyzed by FTIR-MSP in accordance with the protocols
described hereinabove with reference to FTIR-Microspectroscopy.
[0155] The results show that the FTIR-MSP spectral pattern of PBMC
from the cancer patients differs significantly from those of the
healthy controls. The results additionally show that PBMC of the
breast cancer patient and the gastrointestinal cancer patient (with
a history of breast cancer) exhibited a similar spectral pattern
that was distinct from the control. This can be explained by the
gastrointestinal tumor being shown by pathological evaluation to
have a phenotype characteristic of a breast tumor. In some cases of
breast cancer, malignant cells break away from the primary breast
tumor and spread to other parts of the body. These cells may remain
inactive for years before they begin to grow again. It is possible
that a tumor, although located remotely from the original tumor
site, triggers biochemical changes in the PBMC that are similar to
those triggered by the original tumor. It is to be noted that the
spectral absorbance pattern of PBMC of both the breast cancer
patient and the gastrointestinal cancer patient (who has a history
of breast cancer) differ from the spectral pattern produced by PBMC
of patients suffering from other types of solid tumors.
[0156] FIG. 4 shows second derivative spectra of PBMC from the
breast cancer patient and from the gastrointestinal cancer patient
(the gastrointestinal cancer patient having a history of breast
cancer and the gastrointestinal tumor exhibiting a phenotype
characteristic of a breast tumor) compared to PBMC of healthy
controls, as derived in accordance with some applications of the
present invention. As shown, the FTIR-MSP second derivative spectra
exhibit significant differences between PBMC from the cancer
patients and PBMC from healthy controls. These spectral differences
typically represent molecular changes in the PBMC of the cancer
patients and allow distinguishing between the healthy control and
the cancer patients. In addition, PBMC from both the primary breast
cancer patient and the gastrointestinal cancer patient exhibited a
similar FTIR-MSP spectral pattern. Accordingly, some applications
of the present invention can be used to diagnose the type and/or
the origin of solid tumor of a patient based on the unique FTIR-MSP
spectra produced by analysis of the patient's PBMC. This is
consistent with Example 3, which showed that PBMC from patients
with different types of solid tumors each produced a distinct FTIR
spectral pattern with its own set of characteristic bands.
Additionally, the molecular changes which trigger the changes in
the FTIR-MSP spectra of breast cancer patients, including patients
with breast cancer history, can serve as biomarkers to diagnose
breast cancer. Additionally, some applications of the present
invention can be used to select effective treatment based on the
origin of a tumor. It is to be noted that the type of solid tumor
diagnosed in accordance with applications of the present invention
is not limited to breast tumors, but may include any other type of
solid tumors.
EXAMPLE 5
[0157] In this set of experiments, PBMC samples from cancer
patients suffering from various types of solid tumors were analyzed
by FTIR-MSP. The FTIR-MSP spectral pattern of each type of solid
tumor was compared to the FTIR-MSP spectral pattern of the other
solid tumors, allowing distinguishing between different types of
solid tumors. The population of cancer patients for this set of
experiments comprised a total of 63 patients suffering from solid
tumors, in accordance with Table I. The PBMC was obtained by
preliminary processing of the peripheral blood in accordance with
the protocols described hereinabove with reference to extraction of
peripheral blood mononuclear cells (PBMC). The PBMC samples were
then analyzed by FTIR-MSP, in accordance with the protocols
described hereinabove with reference to FTIR-Microspectroscopy.
[0158] The results show that each type of solid tumor produces a
distinct spectral absorption pattern of the PBMC, which is unique
to each type of solid tumor, allowing distinguishing between
different types of solid tumors.
[0159] Reference is made to FIGS. 5A-C. In order to increase
accuracy and achieve effective comparison between PBMC samples of
the various types of cancer patients, the second derivative of the
baseline-corrected, vector-normalized average FTIR absorption
spectra was used. Results are presented in FIGS. 5A-C, showing the
second derivative of several regions of the spectra (the main
absorption bands are marked). As shown in FIGS. 5A-C, the second
derivative spectra of each type of solid tumor produced a distinct
spectral absorption pattern of the PBMC, which is unique to each
type of solid tumor.
[0160] Reference is made to FIGS. 5D-G, which are a series of
graphs representing values of the second derivative of absorption
spectra of each type of solid tumor compared to the second
derivative of absorption spectra of the other solid tumors, derived
in accordance with some applications of the present invention.
[0161] FIG. 5D is a graph representing values of the second
derivative of absorption spectra of PBMC samples from the breast
cancer patients (n=25) compared to PBMC samples from the cancer
patients suffering from other types of solid tumors that do not
include breast cancer (n=38), at wavenumbers B1-B5. Statistical
analysis was performed and P-values are provided. As shown, the
second derivative of PBMC from the breast cancer patients differed
significantly from the second derivative analysis of FTIR-MSP
spectral patterns from PBMC of other cancer patients who do not
have breast cancer.
[0162] Table IV lists the wavenumbers shown in FIG. 5D. Typically,
PBMC samples were analyzed by FTIR-MSP techniques using these
wavenumbers to distinguish between breast cancer patients and
cancer patients who do not have breast cancer.
TABLE-US-00004 TABLE IV Breast vs. Non Breast Wavenumber (cm-1 .+-.
4) B1 752 B2 1030 B3 1046 B4 1128 B5 1237
[0163] FIG. 5E is a graph representing values of the second
derivative of absorption spectra of PBMC samples from the
gastrointestinal cancer patients (n=18) compared to PBMC samples
from the cancer patients suffering from other types of solid tumors
that do not include gastrointestinal tumors (n=45), at wavenumbers
C1-C11. Statistical analysis was performed and P-values are
provided. As shown, the second derivative of PBMC from the
gastrointestinal cancer patients differed significantly from the
second derivative analysis of FTIR-MSP spectral pattern from PBMC
of other cancer patients who do not have gastrointestinal
cancer.
[0164] Table V lists the wavenumbers shown in FIG. 5E. Typically,
PBMC samples were analyzed by FTIR-MSP techniques using these
wavenumbers to distinguish between gastrointestinal cancer patients
and cancer patients who do not have gastrointestinal cancer.
TABLE-US-00005 TABLE V GI vs. Non GI Wavenumber (cm-1 .+-. 4) C1
797 C2 830 C3 893 C4 899 C5 1128 C6 1298 C7 1354 C8 1714 C9 1725
C10 1738 C11 3013
[0165] FIG. 5F is a graph representing values of the second
derivative of absorption spectra of PBMC samples from the lung
cancer patients (n=8) compared to PBMC samples from the cancer
patients suffering from other types of solid tumors that do not
include lung tumors (n=55) at wavenumbers D1-D24. Statistical
analysis was performed and P-values are provided. As shown, the
second derivative of PBMC from the lung cancer patients differed
significantly from the second derivative analysis of FTIR-MSP
spectral patterns from PBMC of other cancer patients who do not
have lung cancer.
[0166] Table VI lists the wavenumbers shown in FIG. 5F. Typically,
PBMC samples were analyzed by FTIR-MSP techniques using these
wavenumbers to distinguish between lung cancer patients and cancer
patients who do not have lung cancer.
TABLE-US-00006 TABLE VI Lung vs. Non Lung Wavenumber (cm-1 .+-. 4)
D1 765 D2 780 D3 797 D4 851 D5 874 D6 881 D7 913 D8 923 D9 958 D10
968 D11 1044 D12 1085 D13 1191 D14 1241 D15 1344 D16 1373 D17 1417
D18 1458 D19 1469 D20 1692 D21 1714 D22 1728 D23 2852 D24 2984
[0167] FIG. 5G is a graph representing values of the second
derivative of absorption spectra of PBMC samples from the prostate
cancer patients (n=5) compared to PBMC samples from the cancer
patients suffering from other types of solid tumors that do not
include prostate cancer (n=58) at wavenumbers E1-E11. Statistical
analysis was performed and P-values are provided. As shown, the
second derivative of PBMC from the prostate cancer patients
differed significantly from the second derivative analysis of
FTIR-MSP spectral pattern from PBMC of other cancer patients who do
not have prostate cancer.
[0168] Table VII lists the wavenumbers shown in FIG. 5G. Typically,
PBMC samples were analyzed by FTIR-MSP techniques using these
wavenumbers to distinguish between prostate cancer patients and
cancer patients who do not have prostate cancer.
TABLE-US-00007 TABLE VII Prostate vs. Non Prostate Wavenumber (cm-1
.+-. 4) E1 828 E2 932 E3 997 E4 1059 E5 1299 E6 1302 E7 1403 E8
1454 E9 1714 E10 2979 E11 3013
[0169] Reference is now made to Example 6. The experiments
presented hereinbelow with reference to Example 6 demonstrate that
in accordance with some applications of the present invention,
analysis of PBMC samples by FTIR-MSP techniques is used for staging
cancer. Typically, each stage of cancer produces distinct FTIR
spectra of the PBMC.
EXAMPLE 6
[0170] In this set of experiments, PBMC samples from cancer
patients suffering from different stages of cancer due to solid
tumors were analyzed by FTIR-MSP and a second derivative of the
average of the spectra was obtained for each of the stages of
cancer, allowing distinguishing between different stages of cancer.
The population of cancer patients for this set of experiments
comprised a total of 63 patients suffering from the different
stages (stages one and two (n=29), and stages three and four
(n=34)) of cancer, as described in Table I.
[0171] The PBMC was obtained by preliminary processing of the
peripheral blood in accordance with the protocols described
hereinabove with reference to extraction of peripheral blood
mononuclear cells (PBMC). The PBMC samples were then analyzed by
FTIR-MSP in accordance with the protocols described hereinabove
with reference to FTIR-Microspectroscopy.
[0172] The results show that early stages of cancer produce
spectral absorption pattern of the PBMC, that are different than
those produced by PBMC samples taken from patients with advanced
stages of cancer, allowing distinguishing among different stages of
cancer, in particular, between early and advanced stages.
[0173] Reference is made to FIGS. 6A-C, which are graphs showing
the second derivative spectra of PBMC and analysis thereof, based
on PBMC samples from cancer patients suffering from different
stages of cancer, derived in accordance with some applications of
the present invention.
[0174] FIG. 6A is a graph representing the second derivative of
baseline-corrected, vector-normalized average FTIR absorption
spectra of PBMC samples obtained from cancer patients in different
stages of the disease. As shown, PBMC of each stage of cancer
produced a distinct spectral absorption pattern of the PBMC.
[0175] FIG. 6B is a graph representing the second derivative of
baseline-corrected, vector-normalized average FTIR absorption
spectra of PBMC samples obtained from breast cancer patients in
different stages of the disease. As shown, PBMC of each stage of
breast cancer produced a distinct spectral absorption pattern of
the PBMC, and particularly, the early stages (stages one and two)
were distinct from the more advanced stages (stages three and
four). It is to be noted that breast cancer is shown by way of
illustration and not limitation, and that the scope of the present
includes staging of any type of solid tumor by techniques described
herein.
[0176] FIG. 6C is a graph representing values of the second
derivative of absorption spectra of PBMC samples from stage one and
two (n=29) cancer patients, compared to stage three and four (n=34)
cancer patients at wavenumbers F1-F7. Statistical analysis was
performed and P-values are provided. As shown, the second
derivative of PBMC from the cancer patients with early stages of
cancer (stages one and two) differed significantly from the second
derivative analysis of FTIR-MSP spectral pattern of cancer patients
with advanced stages of cancer (stage three and four).
[0177] Table VIII lists the wavenumbers shown in FIG. 6A.
Typically, PBMC samples were analyzed by FTIR-MSP techniques using
these wavenumbers to distinguish between early stages (stage one
and two) and more advanced stages (three and four) of cancer.
TABLE-US-00008 TABLE VIII Initial Stages vs. Advanced Stages
Wavenumber (cm-1 .+-. 4) F1 865 F2 897 F3 924 F4 1030 F5 1047 F6
1191 F7 1238
[0178] Reference is made to FIGS. 1-6 and Examples 1-6. It is to be
noted that techniques described herein with reference to use of
peripheral blood mononuclear cells (PBMC) may be applied to any
type of white blood cell (WBC) or a combination of types of white
blood cells. For example, analysis by FTIR microscopy techniques
may be performed on any type of white blood cell, including but not
limited to a total population of white blood cells (e.g., as
obtained by red blood cell lysis).
[0179] Reference is still made to FIGS. 1-6 and Examples 1-6.
[0180] The data obtained by analysis of the PBMC samples may be
further analyzed by any suitable method known in the art, e.g.,
Artificial Neural Network and/or Cluster Analysis, and/or Principal
Component Analysis, and/or Linear Discriminant Analysis (LDA) e.g.,
Fisher's Linear Discriminant Analysis (FLDA), Quadratic
Discriminant Analysis, and/or Non Linear Discriminant Analysis.
[0181] For example, data obtained in accordance with applications
of the present invention may be analyzed by an artificial neural
network (ANN). Several biomarkers shown in Tables II-VII which are
statistically significant (p<0.05) may be served as an input
vector for the ANN analysis.
[0182] It is further noted that the scope of the present invention
includes the use of only one wavenumber (representing one
biomarker) for detection and/or monitoring of a solid tumor, as
well as the use of two, three, four, or more wavenumbers.
[0183] Additionally, the scope of the present invention includes
using any IR spectral feature or any feature derived from analysis
of an IR spectral feature (e.g., any type of peak analysis), to
indicate the presence of a solid tumor.
[0184] It is also noted that the scope of the present invention is
not limited to any particular form or analysis of IR spectroscopy.
For example, IR spectroscopy may include Attenuated Total
Reflectance (ATR) spectroscopy techniques.
[0185] Although applications of the present invention are described
hereinabove with respect to spectroscopy, microspectroscopy, and
particularly FTIR spectroscopy, the scope of the present invention
includes the use of analysis techniques with data obtained by other
means as well (for example, using a monochromator or an LED, at
specific single wavenumbers, and/or FTIR imaging).
[0186] It will additionally be understood by one skilled in the art
that aspects of the present invention described hereinabove can be
embodied in a computer running software, and that the software can
be supplied and stored in tangible media, e.g., hard disks, floppy
disks, a USB flash drive, or compact disks, or in intangible media,
e.g., in an electronic memory, or on a network such as the
Internet.
[0187] It will be appreciated by persons skilled in the art that
the present invention is not limited to what has been particularly
shown and described hereinabove. Rather, the scope of the present
invention includes both combinations and subcombinations of the
various features described hereinabove, as well as variations and
modifications thereof that are not in the prior art, which would
occur to persons skilled in the art upon reading the foregoing
description.
* * * * *