U.S. patent application number 10/293019 was filed with the patent office on 2003-04-17 for breast cancer eradication program.
Invention is credited to Sirbasku, David A..
Application Number | 20030072812 10/293019 |
Document ID | / |
Family ID | 46281529 |
Filed Date | 2003-04-17 |
United States Patent
Application |
20030072812 |
Kind Code |
A1 |
Sirbasku, David A. |
April 17, 2003 |
Breast cancer eradication program
Abstract
A method of treating breast cancer that is at least partially
ER.sup.+ is disclosed. The method comprises administering at a
tumor site in a mammalian subject a pharmaceutically acceptable
form of Fe(II) or Fe(III) in a suitable carrier. A four-part
program aimed at eradicating breast cancer includes (a) local
treatment and prevention of spread from a contained breast site,
preferably using local administration of a ferric iron composition,
(b) treatment of disseminated (metastatic) breast cancer, (c)
reduction in the risk of developing breast cancer, preferably by
enhancing dimeric/polymeric IgA and polymeric IgM inhibition of
estrogen responsive cell growth, and (d) protection against cancer
causing agents.
Inventors: |
Sirbasku, David A.; (Austin,
TX) |
Correspondence
Address: |
CONLEY ROSE, P.C.
P. O. BOX 3267
HOUSTON
TX
77253-3267
US
|
Family ID: |
46281529 |
Appl. No.: |
10/293019 |
Filed: |
November 13, 2002 |
Related U.S. Patent Documents
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Application
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Filing Date |
Patent Number |
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10293019 |
Nov 13, 2002 |
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09852958 |
May 10, 2001 |
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10293019 |
Nov 13, 2002 |
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09852547 |
May 10, 2001 |
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60338037 |
Nov 13, 2001 |
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Current U.S.
Class: |
424/647 ;
514/184; 514/502 |
Current CPC
Class: |
A61K 35/16 20130101;
C07K 16/26 20130101; A61K 2039/505 20130101; G01N 33/574 20130101;
G01N 2333/71 20130101; A61K 2039/542 20130101; A61P 35/00 20180101;
A61K 2039/585 20130101; C07K 14/70567 20130101; G01N 33/96
20130101; C07K 2317/73 20130101; G01N 33/6854 20130101; A61K 31/138
20130101; C07K 16/00 20130101; G01N 33/5011 20130101; G01N 33/743
20130101; C07K 2317/52 20130101; A61K 39/0008 20130101; A61K 33/26
20130101; C07K 16/065 20130101; A61K 2039/545 20130101; C07K 16/12
20130101 |
Class at
Publication: |
424/647 ;
514/502; 514/184 |
International
Class: |
A61K 033/26; A61K
031/555; A61K 031/295 |
Goverment Interests
[0002] Research leading to the present invention was supported in
part by the federal government under Grant Nos. DAMD17-94-J-4473,
DAMD17-98-1-8337 and DAMD17-99-1-9405 awarded by the Defense
Department through the US Army Medical Research and Materiel
Command, Breast Cancer Research Program. The United States
government may have certain rights in the invention.
Claims
What is claimed is:
1. A method of treating breast cancer that is at least partially
ER.sup.+ comprising administering at a tumor site in a mammalian
subject a pharmaceutically acceptable form of at least one iron
compound or complex in a suitable carrier.
2. The method of claim 1 wherein said iron compound or complex
comprises Fe(III) (ferric iron).
3. The method of claim 1 wherein said iron compound or complex
comprises Fe(II) (ferrous iron).
4. The method of claim 1 wherein said administering of said iron
compound or complex is followed by performing surgery to remove
said tumor.
5. The method of claim 1 comprising applying said iron compound or
complex to a mastectomy or lumpectomy site.
6. The method of claim 1 comprising applying said iron compound or
complex to surgical margins of a mastectomy or lumpectomy site.
7. The method of claim 1 comprising administering said iron
compound or complex to a disseminated/metastatic breast cancer
site.
8. The method of claim 1 wherein said iron compound or complex
comprises a soluble or insoluble composition.
9. The method of claim 8 wherein said iron compound or complex
comprises a soluble form of Fe(III) chosen from the group
consisting of ferric ammonium citrate, ferric ammonium sulfate,
ferric chloride, ferric nitrate, ferric sulfate and ferric nitrate
hydroxide, and combinations, complexes or polymeric forms
thereof.
10. The method of claim 8 wherein said iron compound or complex
comprises an insoluble form of Fe(III) chosen from the group
consisting of ferric deferoxamine activated insoluble matrix.
11. The method of claim 10 wherein said activated insoluble matrix
is biodegradable.
12. The method of claim 11 wherein said activated insoluble matrix
is chosen from the group consisting of dextran, starch, insoluble
protein and biodegradable synthetic polymers.
13. The method of claim 10 wherein said insoluble matrix comprises
non-biodegradable implantable beads.
14. The method of claim 1 wherein said iron compound or complex
comprises INFeD.RTM..
15. The method of claim 1 wherein said iron compound or complex
comprises radiolabeled .sup.59Fe(III) or .sup.55Fe(III).
16. The method of claim 1 wherein said iron compound or complex
comprises .sup.59Fe(III) or .sup.55Fe(III) bound to
transferrin.
17. The method of claim 1 wherein said iron compound or complex
comprises a radioisotope of a metal other than iron that is bound
to transferrin or apotransferrin, said radioisotope-bound
transferrin or apotransferrin molecule also being bound to at least
one Fe(III) atom.
18. The method of claim 1 wherein exposure of said tumor site to
said iron compound or complex is no more than a few days.
19. The method of claim 1 wherein said tumor site is other than a
primary localized breast tumor.
20. A method of treating breast cancer comprising ER.sup.- cancer
cells, the method comprising administering to an individual in need
thereof a pharmaceutically acceptable composition comprising means
for deterring or preventing iron uptake by said ER.sup.- cancer
cells.
21. The method of claim 20 wherein said means comprises at least
one chelator chosen from the group consisting of
.alpha.-ketohydroxypyridine iron chelators.
22. The method of claim 20 wherein said means comprises at least
one monoclonal antibody against a transferrin receptor.
23. The method of claim 20 comprising administering said
composition at a tumor site.
24. The method of claim 17 comprising administering said
composition in the blood plasma.
25. The method of claim 24 wherein said composition comprises means
for blocking diferric transferrin from binding to a cellular
transferrin receptor.
26. The method of claim 20 comprising depleting iron in the diet of
said individual.
27. The method of claim 20 wherein said composition comprises a
blood substitute.
28. A method of treating breast cancer containing ER.sup.+ cells,
the method comprising administering to an individual in need
thereof a pharmaceutically acceptable composition comprising means
for increasing the amount of at least one immunoglobulin inhibitor
chosen from the group consisting of IgA, IgM and the Fc domain of
IgA or IgM sufficiently to kill at least a portion of said ER.sup.+
cells in said individual.
29. The method of claim 28 wherein said means comprises oral
challenge with an immunogen capable of causing an increase in the
number and/or function of IgA or IgM secreting B immunocytes in
breast tissue.
30. A method to aid in deterring the occurrence, growth or
progression of breast cancer in a population of susceptible
individuals, the method comprising carrying out at least one of the
following regimens: (a) in individuals having breast cancer or at
risk of developing breast cancer, contacting the breast ductal
tissue of said individuals with a cell growth inhibiting amount of
an immunoglobulin inhibitor or an immunoglobulin inhibitor
mimicking compound; (b) in individuals with ER.sup.+ breast cancer
cells, treating a localized tumor or mastectomy or lumpectomy site
with an effective amount of a pharmaceutically acceptable form of
Fe(II), Fe(III), radioactive Fe(II) or Fe(III), or Fe(III)-bound
transferrin or apotransferrin molecules that are also bound to
another radioactive metal, sufficient to kill at least a portion of
the cancer cells in said tumor or at said mastectomy of lumpectomy
site; (c) in individuals with disseminated ER.sup.+ breast cancer
cells, administering said Fe(II) or Fe(III) to a metastatic cancer
site; (d) in individuals with localized or disseminated ER.sup.+
breast cancer cells, administering locally or systemically a
pharmaceutically acceptable composition comprising means for
increasing the amount of at least one immunoglobulin inhibitor
chosen from the group consisting of IgA, IgM and the Fc domain of
IgA or IgM sufficiently to kill at least a portion of said ER.sup.+
cells in said individual; (e) in individuals with ER.sup.- breast
cancer cells, administering locally or systemically a
pharmaceutically acceptable composition comprising means for
deterring or preventing iron uptake by said ER.sup.- cancer cells;
(f) in individuals with localized or disseminated ER.sup.- breast
cancer cells, treating said ER.sup.- cancer cells with an effective
amount of a pharmaceutically acceptable composition containing
radioactive Fe(II) or Fe(III), or Fe(III)-bound transferrin or
apotransferrin molecules that are also bound to another radioactive
metal, sufficient to kill at least a portion of said ER.sup.-cancer
cells; and (g) in individuals with ER.sup.- breast cancer cells,
infecting at least a portion of said ER.sup.- cancer cells with a
virus vector containing a DNA sequence coding for a poly IgA/IgM
binding receptor, and administering locally or systemically an
immunoglobulin inhibitor of cancer cell growth or a mimic thereof
capable of binding to said receptor.
31. The method of claim 30 comprising carrying out at least one of
the following regimens: (a) in individuals having breast cancer or
at risk of developing breast cancer, enhancing the number of B
immunocytes producing IgA or IgM in breast tissue; and (b)
immunizing individuals at risk of developing breast cancer against
microorganisms known to or suspected of causing breast cancer.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit under 35 U.S.C.
.sctn.119(e) of U.S. Provisional Patent Application No. 60/338,037
filed Nov. 13, 2001, and is a continuation-in-part of U.S. patent
application Ser. Nos. 09/852,958 and 09/852,547, both filed May 10,
2001, the disclosures of each of which are hereby incorporated
herein by reference.
BACKGROUND OF THE INVENTION
[0003] 1. Field of the Invention
[0004] The present invention generally relates to methods and
compositions for the eradication of cancers of the mucosal
endothelial tissues. More particularly, the present invention
relates to the use of such compositions and methods for breast
cancer risk reduction, prevention and treatment.
[0005] 2. Description of Related Art
[0006] For women with breast cancer, the term "eradication" has
different meanings depending upon the state of their disease.
Additionally, for women still disease free, eradication means
preventing the development of breast cancer. Today, there are no
known preventions for breast cancer. Although many risk factors
have been identified for breast cancer (76-78), reduction of
overall risk from the current United States level of one-in-eight
has not been achieved short of the use of the anti-estrogen
tamoxifen as a preventative (79). However tamoxifen is only for use
by very high-risk women (79). It is not for use with the general
population, both because of adverse side effects and because of
disruption of reproductive capacity. Accordingly, tamoxifen is
under evaluation for high-risk women, but is not considered
appropriate for use as a standard preventative due to its serious
side-effects and causation of a number of endocrine problems.
Treatment with tamoxifen for more than five years may in fact
induce breast cancer. It is now recommended that this therapy be
limited to only five years. This does not cover the span of a
women's lifetime. Based on knowledge today, it may be possible to
reduce risk by alterations of life style and diet, but these are at
best fractional gains and offer no assurances. Even women who lead
very health-risk conscious lives still develop breast cancer. A
successful mass prevention is a primary goal in the fight against
this disease.
[0007] For women diagnosed with breast cancer, the current most
effective eradication methods begin with various surgical
procedures (i.e. mastectomy or breast conservation surgery or more
commonly lumpectomy). Without doubt, removing a primary tumor is an
effective first step of eradication provided tumor cells have not
escaped to other body sites. Given that we understand that escape
is a possibility, or that multiple unrecognized tumor foci are
present in one or both breasts, many breast cancer patients opt for
radiation therapy, adjuvant chemotherapy and/or tamoxifen treatment
even when they are diagnosed as node negative. The term "node
negative" indicates that cancer cells have not moved to the
axillary nodes from the breast. Commonly however, it is thought
useful to "shrink" primary tumors before surgical removal. Today
this is done by systemic chemotherapy or radiation therapy.
Patients with node negative estrogen receptor positive (ER.sup.+)
breast cancer may also be treated with anti-estrogen (e.g.
tamoxifen) as is often done for postmenopausal women. Depending
upon age and physical condition, anti-estrogen therapy is now a
common alternative for many postmenopausal women.
[0008] For breast cancer patients who are axillary node positive
(either ER.sup.+ or ER.sup.-), additional treatment is essential
including those noted above. There is no doubt that these
modalities have a major positive impact on long term survival, but
it is likewise clear to basic cancer investigators and cancer
clinicians alike that the failure rate is significant even despite
frequent quoting of favorable statistical data. Currently, node
positive women without other evidence of dissemination can be
treated by standard chemotherapy and/or radiation therapy. However,
node positive patients with ER.sup.- tumors are at risk no matter
the therapy utilized. For these women, the use of anti-hormone
therapy is not as effective as with patients with ER.sup.+
tumors.
[0009] It is with the diagnosis of metastases to liver, bone,
brain, lung, etc., that another more serious level of the
eradication issues arises, and standard chemotherapy is most often
not effective. For reasons both known and unknown, these cases have
a very poor prognosis. Current chemotherapy can sometimes retard
metastatic cancer growth, but as cancers spread they become
progressively more therapy resistant. The level of public concern
about this issue is clear from anticipation of the benefits of such
new drugs as Herceptin.RTM., which is a monoclonal antibody against
the HER2 receptor. Introduction of Herceptin.RTM. was accompanied
by widespread reports in the news media that heightened
expectations. Unfortunately, even this new family of
biopharmaceuticals is not an effective means of eradication. At
best, Herceptin.RTM. provides only a small increase in survival
time and then only in combination with chemotherapy and only with a
minority of treated patients (70). While other types of drugs are
under investigation, the magnitude of the disseminated cancer
problem remains undiminished.
[0010] Iron deprivation has been discussed as a means of
eliminating cancer cells (39,40), but the focus has been on two
technologies that, used alone, have not worked. First, those
investigators have considered iron deprivation via treatment with
chelators that bind the metal and thereby render it metabolically
inactive. Chelation alone has not removed enough iron from the body
to be effective as an anticancer program (41). This is to be
expected, for it is known that iron is retained in organs and
tissues with a biological half-life of about 2000 days. Most
likely, chelation alone will not be an effective therapy.
[0011] Currently, the primary effort in breast cancer research
aimed at eradication of the disease is intensely focused by
powerful technology that permits identification of a large number
of genes, and by the human genome project that promises to solve
the cancer problem. To date, these technologies have provided
valuable information but have failed to move to the next level of
application, cancer eradication.
[0012] A brief historical overview of breast cancer research over
the past four decades points to numerous periods of advancement,
each with its own promise of defeating cancer. For example, during
the 1960s investigators were encouraged by newly identified enzyme
and metabolic changes in cancer cells. It then seemed clear that
these changes were the cause of cancer and that its end was near.
During the 1970s, the beginnings of molecular biology (then called
microbial genetics) yielded new found information that was thought
certain to lead to the end of many human diseases, including
cancer. Investigations in the field of endocrine cancer research,
during the 1980s, focused on how hormones caused cell growth and
developed animal models to study hormone dependent cancer. At that
time, serum-free defined animal cell culture was being developed
(1) and new substances called growth factors were being explored
(2). Also during that period, another major advancement was the
discovery of the estrogen receptor (ER) and the hypothesis that it
alone controlled estrogen dependent cell growth (3-5). Some
investigators did not accept all of the ER hypothesis (6-8),
however, and thought that estrogen-inducible growth factors
(estromedins) were necessary (6,9,10). It appeared clear at that
time that growth factor research would untangle the cancer enigma.
Today cancer scientists know this is not the case.
[0013] Along with the growth factor research came the "oncogene"
explosion of the 1990s, which promised an end to cancer. Today,
cancer investigators are inundated by scores of gene changes in
cancer. The list grows weekly. A GENBANK search of "breast cancer
hot spots" yielded more than 100 "hits" on several chromosomes.
This cornucopia of genetic information obscures two facts: First,
very few breast cancers can be traced to germ line DNA changes
(11). Most are not inherited. Notable exceptions are BRCA1 and
BRCA2, which represent at most 1-10% of breast cancers in the
United States (31-33). Given that the incidence of breast cancer
now approaches 1 in 8, the majority of breast cancers have other
origins. Second, sophisticated new molecular technology has
identified changes in expression of at least 100 mRNAs in breast
cancer cells (11). There is promise of hundreds of gene/expression
changes (11-13). It is very unlikely they are all causative or even
critical to breast cancer. The tempting scenario is to investigate
each mRNA or gene to define its role. Of course, this represents
years of work for researchers, and still leaves open the question:
"Will this lead to breast cancer eradication"?
[0014] It is known that eighty percent or more of breast cancers
are invasive ductal carcinomas that arise from ductal cells (85,86)
or precursors of ductal cells (85,87). Based on the current state
of knowledge, there is no genetic lesion to explain the 70% of
breast cancers now termed "sporadic". Certainly the BRCA1 and BRCA2
genes are responsible for at most a small percentage of breast
cancers in this country (88-91). Lesions in the p53 gene were
initially thought to be important in as many as 15 to 50% of breast
cancers (92-94). However, it was far from clear which mutations are
causally related to breast cancer onset or which actually
constitute secondary changes leading to loss of function of this
tumor suppressor gene in the different types of breast cancer (i.e.
ER.sup.+ or ER.sup.-). A more recent study has rightly pointed out
the confusion regarding p53 mutations and breast cancer patients
(96). Studies of p53 mutations have yielded a wide range of results
depending upon the methods employed (97). One useful fact is that
the results average about 30 to 40% for loss of heterozygosity at
the p53 gene (97). This means the remaining gene may be a "hot
spot" (i.e., a chromosomal loci or gene that is frequently altered
in breast cancer specimens). However, at this time, there is not
sufficient evidence to support the use of p53 as a guide to
selection of therapy modalities for breast cancer (98).
[0015] In fact, today it is very difficult to explain the great
many mutations and other types of genetic expression alterations
that are known in breast cancer cells (11). Based on the findings
with breast and other types of mucosal cancers, such changes
include mutations, translocations, amplifications of oncogenes,
loss of heterozygosity (LOH), and allelic imbalances
(12,13,100-102). How do all of these happen? Are environmental
carcinogens in such high abundance that they explain these data?
Despite the concentrated focus given to environmental carcinogens
as causes of breast cancer (20,95,99), that hypothesis has failed
to move forward to the level of accepted scientific fact. Ways to
reduce the risk of developing breast cancer, ways of preventing its
occurrence, and ways to treat existing cases of localized and
metastatic breast cancer are urgently needed. Even with the very
best of treatments currently available, a longer-term plan is still
needed in which prevention is the first line of eradication. A
successful prevention will be, preferably, safe and have low or
negligible side effects. It should be capable of reducing risk for
the majority of women, independent of their economic circumstances.
It should cause little or no disruption of life-style or
reproductive capacity.
SUMMARY OF PREFERRED EMBODIMENTS
[0016] While continued gene searching may not lead to the goal of
eradication of breast cancer in the near or even mid-range future,
the present invention offers eradication technology that can be
applied today--no matter how many gene changes are ultimately
associated with cancer development. Eradication is approached from
a unique perspective based on discoveries described in more detail
below and in co-pending U.S. patent application Ser. Nos.
09/852,547 entitled "Compositions and Methods for the Diagnosis,
Treatment and Prevention of Steroid Hormone Responsive Cancers" and
Ser. No. 09/852,958 entitled "Compositions and Methods for
Demonstrating Secretory Immune System Regulation of Steroid Hormone
Responsive Cancer Cell Growth", and in corresponding International
Patent Applications PCT/US01/15171 (WO 01/86307 and PCT/US01/15183
(WO 01/85210), also identified in the list of References, below, as
items 29 and 30, and hereby incorporated herein by reference).
[0017] The present invention specifically focuses on the
eradication of breast cancer, and overcomes many of the problems
and barriers in breast cancer research today. In certain
embodiments of the invention, (ferric) iron-based treatment of
local breast cancer tumors and lumpectomy sites is provided. New
treatment strategies for three conditions are disclosed, which
include (i) treatment of mastectomy sites to eliminate residual
cancer cells, (ii) treatment of primary tumors before surgery,
(iii) treatment of the surgical margins of mastectomy sites to
eliminate undetected residual cancer cells.
[0018] In certain embodiments of the invention, treatment of
disseminated/metastatic breast cancer is addressed from a cell
nutrition perspective. Both ER.sup.+ and ER.sup.- metastatic breast
cancers are highly growth dependent upon diferric transferrin as a
source of metabolic iron required for cell growth (29,30,71,72) and
more specifically DNA synthesis (73,74), as described in the above
cited USPTO pending patent applications, using newly developed
serum-free medium cell culture methods. Because of this strict
requirement for diferric transferrin, manipulation of iron
metabolism is employed to kill disseminated cancer cells.
[0019] In certain embodiments of the invention, risk reduction via
oral "immunization" is provided, i.e., oral administration of
immunogens that result in increased content of secretory
immunoglobulins (IgA and IgM) in breast tissue. This approach to
risk reduction is based on the very well established fact that DNA
synthesis (i.e. cell replication) is required to achieve the full
effects of mutagens (80-85). The secretory immunoglobulins IgA and
IgM are inhibitors of breast cell DNA synthesis (29,30) and
therefore reduce the probability of mutations that lead to breast
cancer later in life.
[0020] Certain embodiments of the present invention provide methods
for eradicating breast cancer by conventional oral or standard
immunization against bacteria or other microorganisms existing in
the breast duct system that release or cause formation of mutagenic
agents that lead to causative genetic changes in the exposed ductal
cells. This approach addresses the problem of what single source
might give rise to a process that can cause so many mutations and
different genetic changes that accumulate over the known prolonged
period required to develop breast cancer. Identification of the
causative bacteria/microorganisms makes possible the exploitation
of the body's secretory immune system to develop secretory immunity
or to use standard immunization to transmit immunity to the ductal
fluids. Protection from the underlying causative agents will
provide the best means of ultimate eradication of breast cancer. A
microbial origin of breast cancer does not appear to have been
previously described or suggested in the scientific literature.
[0021] In preferred embodiments of the present invention, all four
parts of the breast cancer eradication program are applied to
appropriate groups of affected or at-risk individuals, including
(1) local treatment and prevention of spread from a contained
breast site; (2) treatment of disseminated (metastatic) breast
cancer; (3) reduction in the risk of developing breast cancer; and
(4) protection against cancer causing agents. In some embodiments,
one or more parts of the program are employed for treatment,
reduction of risk, or prevention of breast cancer in a single
individual or a one or more groups of individuals. Full
implementation of the preferred four-part integrated program is
expected to eradicate breast cancer within the next decade. These
and other embodiments, features and advantages of the present
invention will become apparent with reference to the following
description.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
[0022] The breakthrough in breast cancer research disclosed in U.S.
patent application Ser. Nos. 09/852,958 and 09/852,547, and which
also applies to cancers of other mucosal epithelial tissues in the
body, is further implemented and extended herein. The disclosures
of those applications are hereby incorporated herein by reference.
Purification of new "serum factor(s)" that regulate estrogen
responsive breast cancer cell growth in culture is described in
those preceding applications. The purification yielded
dimeric/polymeric immunoglobulin A (IgA) and pentameric
immunoglobulin M (IgM) as the active regulators. These
immunoglobulins ("immunoglobulin inhibitors") arrested estrogen
target tumor cell growth completely at low nanomolar
concentrations, and their inhibitory effects were entirely
reversible by picomolar concentrations of estrogens. That
disclosure revealed a previously unknown function for the secretory
immune system. In the above-identified patent applications, a major
role for TGF.beta. in breast growth regulation is also identified:
it is a cytokine that controls IgA/IgM immunocytes. Breast cancer
growth is best defined as negative paracrine control by secretory
immunoglobulins (immunoglobulin inhibitors) and positive direct
control by estrogens. In conjunction with this work, the
longstanding problem of the regulation of estrogen dependent cell
growth in culture under serum-free defined medium conditions was
solved. These results have great physiological relevance. IgA and
IgM are secreted by B immunocytes located in the lamina propria of
estrogen target tissues including breast. They are more than 90% of
the immunoglobulins secreted into breast milk. The positioning of
the immunocytes in the tissue adjacent to the epithelial cells and
the secretion of the immunoglobulins is hormone regulated.
[0023] In the course of developing suitable serum-free defined
culture media for studying estrogen effects on breast cancer cell
growth, it was discovered that both soluble iron (FeIII) and
diferric transferrin had special roles with regard to estrogen
receptor positive (ER.sup.+) and ER.sup.- breast cancer cell
growth. Even more surprising, the results point to a new estrogen
receptor ("ER.gamma.") with a higher affinity for steroid hormone
than that of the known receptors ER.alpha. and ER.beta.. In
addition, this work led to the identification of a mediating
receptor for IgA/IgM that shares the properties of the classical
immunoglobulin transcytosis Poly-Ig receptor and is an Fc receptor
superfamily member. This receptor maps to a gene location linked to
allelic imbalances in 75% of breast cancer specimens. These
discoveries lend themselves to major advancements in breast cancer
eradication, which are further developed and described herein in a
four-part program that has been devised to achieve the goal of
eradication of breast cancer. This four-part breast cancer
eradication program discloses specific new solutions to the four
most pressing aspects of breast cancer eradication:
[0024] (1) Local treatment and prevention of spread from a
contained breast site
[0025] (2) Treatment of disseminated (metastatic) breast cancer
[0026] (3) Reduction in the risk of developing breast cancer
[0027] (4) Protection against cancer causing agents
[0028] For the nearly 180,000 women in the United States who will
be diagnosed with breast cancer this year, the most important issue
is eradication. For some women, eradication means eliminating
existing localized disease. Accordingly, in Part I, below, a new
non-toxic method for treatment of localized cancers is based on
direct application of a soluble form of iron that kills early forms
of cancer. Another approach is direct "immuno-therapy" with
immunoglobulin inhibitors of cancer cell growth. For other women,
eradication means destroying cancer that has moved from the breast
to other locations in the body, i.e., disseminated or metastatic
breast cancer. Today, even with the best treatments metastatic
disease has a poor prognosis. In Part II, below, a number of new
therapeutic approaches are presented herein, which exploit cellular
nutritional requirements for growth of breast cancer cells. For
disseminated breast cancer, a different aspect of iron metabolism
is exploited to kill the tumor cells. In addition, the use of a
new, previously unrecognized breast cancer gene identified in U.S.
patent application Ser. Nos. 09/852,958 and 09/852,547/PCT
Published Application Nos. WO 01/86307 and WO 01/85210 as a poly-Ig
(Fc) receptor or a poly-Ig-like (Fc) receptor that mediates IgA/IgM
inhibition of cancer cell growth is employed in the present
eradication program. Until now, the only breast cancer genes known
were BRCA1 and BRCA2. The problem has been, however, that BRCA1 and
BRCA2 are important for only 1 of 400 women in this country. This
leaves 70% or more of breast cancers with no known genetic origin.
The new gene shows "allelic imbalances" in 75% of breast cancers.
It is a likely candidate for the 70% of breast cancers not
explained today. The new breast cancer gene, coding for the poly-Ig
(Fc) receptor or poly-Ig-like (Fc) receptor is a potentially
valuable candidate for a new gene therapy for disseminated breast
cancer because it can restore immune inhibition and anti-estrogen
inhibition to a cell and can even lead to cancer cell death.
[0029] The surest means of breast cancer eradication is prevention,
which is the focus of Part III, below. The Inventor's studies in
cell culture have proven that the immunoglobulins IgA and IgM from
the secretory immune system can serve to kill early breast cancer
cells by terminally arresting their growth. It is now proposed that
oral challenge will be successfully employed to reduce the risk of
cancer causing mutations in breast cells, as described in more
detail in Part IV, below. Immunity is transferred from the gut to
the breast via the secretory immune system. This strategy is
consistent with the fact that breast cancer incidence is lowest in
areas of the world where oral immune challenges are common and
highest in the Western countries where similar challenges are
restricted. The "hygiene hypothesis" that we are "too clean" will
be applied to reduce the current Western world risk of breast
cancer (1 in 6-8) to the 1 in 40-100 rate of the non-Western
world.
[0030] In another view, prevention can mean full eradication
worldwide. As proposed in U.S. patent application Ser. No.
09/852,547/PCT Published Application No. WO 01/86307, cancer
causing agents are located in the ducts of the breast gland and
infectious agents are responsible for the development of breast
cancer. It has been known for many years that the majority (i.e.
75%) of breast cancers arise from the cells lining the ducts of the
gland. In light of the present disclosure, this is an extraordinary
clue to the cause of breast cancer. Microbes and possibly viruses
inhabit the milk ducts. Furthermore, there are strong recent clues
that cancers may be of microbial origin by virtue of their
metabolic products or secreted proteins. The causative organisms
will be sought from breast tissue specimens as well as human milk.
Conventional-type oral immunizations will be employed to kill the
culprit organisms in the ducts. This approach is consistent with
other time-tested methodologies, such as Sabin's oral vaccination
against the poliomyelitis virus, which is based on the same
secretory immune responses that, in the present case, are directed
against breast cancer causing bacteria. Alternatively, standard
inoculation immunizations with modified microorganisms or fragments
of the suspect microbes will be employed to elicit production of
natural immunoglobulin inhibitors of cancer cell growth. No matter
the general or technical approaches that are ultimately successful,
immunizations have been used worldwide to eradicate human diseases.
A similar attack is expected to bring an end to breast cancer for a
majority of women.
Part I: Local Treatments that Prevent Further Spread from a
Contained Breast Site
Iron Effects on Local Breast Tumors
[0031] Methods and compositions for treating localized breast
cancer involve iron effects on local breast tumors, and employ
nutritional information developed from previous studies of the
differential roles of iron metabolism in steroid hormone receptor
positive (ER.sup.+) and steroid hormone receptor negative
(ER.sup.-) breast cancer cells. These studies were done with
well-known cell lines (29,30,110) grown under serum-free defined
conditions (29,30,110) that permitted precise control of free iron
(FeIII) and diferric transferrin concentrations (21-30). In U.S.
patent application Ser. Nos. 09/852,958 and 09/852,547/PCT
Published Application Nos. WO 01/86307 and WO 01/85210 the need to
reduce free or soluble Fe(III) concentrations to very low levels to
achieve sex steroid hormone or thyroid hormone dependent epithelial
cell growth in culture is disclosed, and such disclosure is hereby
incorporated herein by reference.
[0032] For in vivo treatment of localized breast cancer, preferred
forms of iron used and the preferred compositions are described as
follows: The most cancer cell toxic forms of soluble Fe(III)
presently identified are complexes of ferric ammonium citrate
(about 16% iron by weight) and ferric ammonium sulfate. These solid
salt mixtures are dissolved in water at high concentrations
(1.0-250 mg/ml), and after filter sterilization, and preferably
used immediately. These mixtures are light sensitive. Mixtures
stored at 4 to 27.degree. C. under normal room light conditions
(for example, 3 to 5 days or up to 30 days) show an increase in
cytotoxicity of 2 to 10-fold. Higher intensity lights are also
effective, using wavelengths ranging from low ultraviolet to
visible. Addition of sodium chloride at 0.001 to 0.5 M facilitates
the light sensitivity. Addition of up to 0.01 M sodium phosphate,
pH 7.0, has a similar toxicity enhancing effect provided it does
not cause precipitation. More acidic pH is also effective. More
basic pH precipitates the iron complexes. Elevated temperatures to
60.degree. C. increased toxicity. Longer-term storage of up to 30
days increased toxicity in the presence of light or in light
shielded containers. It is preferred to use the iron composition
immediately after preparation for maximum consistency, particularly
when handled under surgical conditions, and to provide a more
uniform base for testing efficacy. However, in some situations of
use the long shelf-life of the iron compositions is advantageous,
and the amount administered can be adjusted for the increased
potency of the stored composition. Ferric chloride, ferric nitrate
and ferric sulfate were effective, but less so than the
iron-ammonium-citrate complexes. This is likely due to greater
solubility of the iron-ammonium-citrate complexes. Still other
suitable compounds or complexes of ferric iron may exist that also
have the desired cancer cell toxicity properties, and it is
expected that one or more of those forms of Fe (III) could be
substituted for the above-identified forms. In addition to the
ferric iron compositions, there are also ferrous iron compositions
(e.g., ferrous ammonium citrate) that also kill cancer cells when
applied immediately after preparation to a localized breast cancer
site, or a mastectomy or lumpectomy site. Preferably the ferrous
salt is dissolved fresh and used immediately, as air (oxygen)
converts Fe(II) to Fe(III) very quickly. Without wishing to be
limited to a particular theory, it is believed that the Fe(II)
(ferrous) compounds shower the cancer cells with oxidative products
that cause cell death as the Fe(II) is converted to the Fe(III)
form. The Fe(III) conversion product then operates to provide
further cell killing effects, as described above.
[0033] In an in vitro model cell culture system as described in
U.S. patent application Ser. Nos. 09/852,958 and 09/852,547/PCT
Published Application Nos. WO 01/86307 and WO 01/85210, exposure of
ER.sup.+ breast cancer cells to sufficient iron causes cell death
in .ltoreq.48 hours. ER.sup.- breast cancer cells are insensitive
to Fe(III) killing under similar in vitro test conditions. These in
vitro model systems are believed to be predictive of the in vivo
effect of iron treatment on ER.sup.+ breast cancer cells.
[0034] Another alternative form of iron administration for killing
localized cancer cells is prepared by adding .sup.59Fe(III) to one
of the unlabeled (non-isotope) ferric or ferrous salts described
above to increase effectiveness. The radioactive and
non-radioactive iron can be prepared as the same salt or the
admixture of two or more salts of Fe(III). The effectiveness of the
composition is increased because any tumor cells not reached by the
soluble iron will be exposed to DNA fragmenting .gamma. radiation
via .sup.59Fe that penetrates at greater distances.
[0035] One other polymeric form of Fe(III) has high
inhibitory/toxic activity. The combination of ferric nitrate and
bicarbonate is prepared chemically as described (75). After
permitting precipitation of ferric hydroxide, and its removal, the
polymeric form of ferric-nitrate-hydroxid- e remains in clear
solution indefinitely at room temperature. It is used in this clear
solution form.
[0036] A pharmaceutical (FDA approved) iron dextran product
(INFeD.RTM., Watson Pharmaceuticals, Inc., Los Angeles, Calif.) is
available today (137) that has the properties of one suitable
preparation for non-radioactive iron treatment of localized breast
cancer. It is supplied as a single injection 100 mg dose of iron in
2.0 ml. A series of warnings are supplied concerning the use of
INFeD and precautions to be taken with this preparation. There is
an indication that animals treated with high doses may develop
cancers after repeated injections, however the incidence of such
complications in humans is either not clear or very low. However,
it is clear that in the older literature sarcomata were found at
the injection sites of intramuscular iron (158-160). Evaluation of
the literature reveals that sarcomas at the site of iron dextran
injection are (i) species specific, with rodents most likely to
develop tumors (156), especially after repeated injections (157),
(ii) dose and threshold dependent, (iii) residual characteristics
at the site, and (iv) latent period relative to the life span of
the test species. As reviewed in 1977, introduction of
intramuscular iron therapy more than 22 years before then resulted
in only nine malignancy reports in man. Of these, only one appears
causally related to the iron injections (156). Care must be given
to various aspects such as methods of delivery, dilution of drug,
and overall iron status of the patients (161). Attention must be
given to anaphylactic reactions (161). Current use of intramuscular
iron preparations with proper techniques in humans does not appear
to bear a significant risk (161). The iron in the preparation
associates with serum ferritin or hemosiderin, and to a lesser
extent with apotransferrin (i.e. transferrin without bound iron).
Care is taken to monitor serum ferritin levels to determine
potential iron overload. With intramuscular injection, the majority
of the iron dextran is absorbed in 72 hours. The remaining iron
dextran is absorbed over the next 3 to 4 weeks. This rate of
absorption is compatible with use as a treatment for local breast
cancer. For additional cancer cell killing capability,
.sup.59Fe(III) can be incorporated into the iron dextran to
increase effectiveness, as discussed above with respect to it use
in iron salt solutions.
[0037] In still another way in which iron can be used to effect
tumor cell death, diferric transferrin is prepared radio-labeled
with .sup.59Fe(III) or .sup.55Fe(III) (132,133) and is used to
irradiate breast cancer cells to cause cell death. Each
apotransferrin molecule accepts two Fe(III). These bind at neutral
pH with high affinity (i.e. K.sub.a=10.sup.20) to similar "N" and
"C" lobes of the transferrin molecule (136). When breast cancer
cells are grown in serum-free cell culture medium containing 0.01
to 20 .mu.g/ml of .sup.59Fe-transferrin, the radioactive iron from
the transferrin becomes incorporated into many cellular components.
Subsequent radiation induced DNA damage leads to cell death within
24 to 168 hours. This effect was seen with both ER.sup.+ and
ER.sup.- breast cancer cells grown in serum-free defined culture
medium. Other radioisotopes such as .sup.131I-transferrin and
.sup.125I-transferrin (132,133) as well as several other
radioisotopes of metals that bind to apotransferrin can be expected
to serve an equivalent function. The use of .sup.59Fe is preferred
because of its 44.6-day half-life that is suitable for radiation
therapy effectiveness. Yttrium-90 is also a consideration because
of its high-energy .beta. emission and 64.1 hour half-life. Of the
two iodine isotopes, each has advantages. The .sup.125I has a
longer 60-day half-life compared to .sup.131I with an 8-day
half-life. The .sup.131I has higher energy. The advantages of the
different isotopes in killing of ER.sup.+ and ER.sup.- breast
cancer cells will be evaluated first in cell culture and then using
in vivo rat mammary tumor induction models as described herein.
[0038] Another alternative method and composition employs an
insoluble form of radio-labeled iron, which is prepared by methods
previously described (27). When it is necessary or desirable to
limit the amount of radiation to a restricted site, such as with
mastectomy or lumpectomy, or to a specific set of axillary nodes,
the .sup.59Fe can be delivered via a complex of
deferoxamine-Sepharose. Deferoxamine is covalently attached to
Sepharose (27), or to any other "activated" insoluble matrix (human
compatible) by the methods described or derivative methods.
Deferoxamine is a low molecular weight bacterial product that is
currently in use as DESFERAL.RTM. (Novartis Pharmaceuticals Corp.,
East Hanover, N.J.) to treat human iron overload patients (138).
Deferoxamine binds Fe(III) with a very high affinity (i.e.
10.sup.23). The complex of deferoxamine-Fe(III) does not dissociate
under body conditions. Thus, placement of insoluble deferoxamine
bound .sup.59Fe in any site will effectively expose a local area to
high-energy .gamma. radiation with escape of only minimal labeled
Fe(III).
[0039] It may be preferable to select a biodegradable matrix (e.g.
dextran, starch or insoluble protein or biodegradable polymer) in
some cases or a stable/non-degradable matrix (e.g. cellulose or
synthetic biomatrix) in others. Another application includes
attachment of the deferoxamine to non-degradable "biobeads" for
implantation directly into the local tissue for specific periods of
time. The visible non-immunogenic beads can be removed at times
deemed desirable or when the desired effect has been achieved, or
they can be removed at the time of mastectomy or lumpectomy.
[0040] Localized Breast Cancer Eradication. In earlier studies of
the role of nutrients, hormones and growth factors in hormone
responsive pituitary tumor cell growth in serum-free chemically
defined culture, it was observed that 1 .mu.M soluble iron in the
form of Fe(III) inhibited growth (21-28). Exposure to 10 .mu.M
Fe(III) killed these cells. The results were thought initially to
be applicable only to rat pituitary cells. However, they have
proven useful with ER.sup.+ breast cancer cells (29,30). To develop
in vivo confirmation that iron can be used to locally treat breast
tumors, a series of experimental animal models will be
investigated.
[0041] The use of iron for the treatment of cancer is a clear
departure from the widely held belief or paradigm that Fe(III)
cannot be (or should not be) administered locally in vivo. It is
commonly cited (87) that Fe(III) released from cellular ferritin
induces (.cndot.OH) free radical formation and that this reactive
species modifies proteins, lipids and nucleic acids (120). Thus,
investigators generally view iron as cancer initiator or promotor
(87,121-125). That paradigm is not pertinent to the present
therapeutic forms of Fe(III), however, because in the present case
the metal will act only short term. The Fe(III) applied to the
tumors is extracellular and has little or nothing to do with the
complex models developed for explaining the putative role of
intracellular ferritin H chain in oxidative damage to cells
(120,121). Notably, as is indicated (121), much of the ferritin
oxidative model is presumptive and unsubstantiated. Furthermore,
ER.sup.+ breast cancer cells appear to be exquisitely sensitive to
a putative burst of extracellular oxidative products. These cells
die very quickly when non-protein bound Fe(III) is added to culture
medium. It should be noted that free Fe(III) does not support
epithelial growth. Diferric transferrin is required. For cancer
treatment, the period of exposure in vivo will be limited by the
fact that within a few days the Fe(III) will be converted to the
inactive but metabolically useful forms of monoferric and diferric
transferring and ferritin. Free/soluble Fe(III) is expected to bind
to apoferritin and apotransferrin under physiologic conditions.
Plasma contains about 2 mg/ml of apotransferrin and 1 mg/ml
diferric transferrin (i.e. transferrin is 66% unsaturated with iron
in plasma). Since Fe(III) cell killing happens in less than a few
days, the risk of other adverse effects of the iron are minimized.
Certainly long-term mutagenic effects are minimized. The time of
exposure and dose schedule of free Fe(III) will be kept to the
minimum needed to achieve therapeutic results. This is the same
principal used with short doses of .gamma. radiation and short-term
applications of chemotherapy used today to treat breast cancer. In
fact, the very same argument can be made against the radiation
protocols used today to treat localized breast cancer, which run
contrary to the paradigm that excess radiation can induce tumors.
Likewise, several of the current chemotherapy chemicals are
actually mutagenic. Therefore, they are used in regimens that kill
tumor cells but stop short of causing a substantial increase in
other cancers.
[0042] Further support for the value of this approach comes from
the Physician's Desk Reference information (137) discussed above,
in which humans are treated with intramuscular injections of iron
dextran (INFeD) to correct iron deficiencies that are not treatable
by oral therapy. While there have been individual reports of the
appearance of sarcoma tumors at the injection site in humans (137),
such reports could not be confirmed by the manufacturer at the
present time (personal communication with Watson Pharmaceuticals,
Inc.). The frequency of such tumors as a proportion of the total
injections per year or patients treated per year is not available,
but is presumed to be very low.
[0043] The use of systemic iron or orally administered iron causes
an increase in the body content of this metal and in plasma
ferritin and diferric transferrin levels. One report states that
increased dietary iron facilitates carcinogen induction of rat
mammary tumors and estrogen induction of Syrian hamster kidney
tumors (139). Another report (140) states that excess iron again
appeared to facilitate carcinogen induced rat mammary tumors, but
there was more care given to control the effects of various iron
status states on body weight gain and hematocrit. The effects of
excess iron were only apparent later in that study. In another,
study, support for a critical role of iron was not found with the
rat mammary tumor models (141). In the present case, it is
concluded that increased saturation of apotransferrin by dietary
iron results in greater growth rates in carcinogen induced rat
mammary tumor cells. This is consistent with a previous showing
with a carcinogen induced rat mammary tumor cell model in culture
that diferric transferrin is absolutely required for growth
(142,143). Apparently, the systemic elevation of plasma iron is
conducive to growth of breast cancer cells. Any therapy with
Fe(III) for treatment of breast cancer is therefore, preferably
local and is subject to natural elimination within a period of a
month. Preferably the doses are managed such that they do not
substantially elevate plasma ferritin or the iron saturation
percent of transferrin.
[0044] Today, women with localized breast cancer have two initial
surgical options: mastectomy or breast conserving surgery as known
as "lumpectomy". With increasing frequency, pretreatment is done to
shrink primary or nodal tumors before surgery. According to the
present plan, an animal model will be used to test whether iron in
the form of soluble ferric ammonium complexes can destroy existing
tumors, or can eliminate undetected cells within
mastectomy/lumpectomy sites. Initially, this program will include
testing the direct effect of Fe(III) on estrogen growth responsive
tumors developed from rat mammary MTW9/PL2 cells in W/Fu female
rats (34,35). Studies will test treatment by application directly
into tumors or into their immediate blood supply. Additionally,
after tumors have developed they will be resected and the surgical
site treated with soluble Fe(III) to determine effect on
recurrence. It is known that without any treatment, there is a 40
to 60% recurrence rate in four months in rats.
[0045] With a different model based on CD-rats, environmental
carcinogens will be used to induce rat mammary primary tumors as
described (36) before initiating localized Fe(III) treatment. The
primary carcinogen induced rat model selected has many
characteristics of human breast cancer (37) and therefore is
considered relevant.
[0046] Another model also has special relevance. It is now clearly
established and almost universally accepted that estrogens promote
target tissue cell growth (109). There is still a question about
the exact DNA and functional sequence of the receptor that mediates
this response (29,30). However, these steroids may have a second
function. Investigators have long proposed that estrogens (or their
metabolites) are genotoxic and cause mutations (107,108). Estrogens
are considered central to human female breast cancer development
even beyond their growth promoting function. There is a rat mammary
tumor model that mimics this dual effect. Estrogen
(17.beta.-estradiol) treatment of female ACI rats induces mammary
tumors in 100% of the population within 197 days (49). The tumors
are estrogen growth responsive. This model will also be used to
induce tumors and determine the effects of iron therapy on the
primary neoplasms. Positive results with this model will have
special applicability to human cancers. Local treatment with
Fe(III) provides an entirely new first line of eradication of
breast cancer.
[0047] In parallel studies, the animal tumor models described above
will be injected or otherwise treated with the
.sup.59Fe-deferoxamine-Sepharos- e complex and the effects on tumor
mass monitored. This same procedure will be assessed for its effect
on recurrence of resected tumors. This is believed to be a
completely new approach to local breast cancer eradication. The
effects of soluble Fe(III) versus those of immobilized .sup.59Fe
will be compared for tumor regression, survival of the hosts and
effects of both treatments on the physiological health of the
animals. Confirmation obtained in these in vivo rodent studies will
indicate the applicability of, and will supply partial evidence for
FDA approval for, human trials. Today, the only other direct local
breast cancer treatment without systemic effects is radiation,
which causes healing problems post-surgery and other chest wall and
organ complications.
Part II: Treatment of Disseminated (Metastatic) Breast Cancer
Methods of Treating Disseminated Breast Cancer
[0048] The problem of eliminating disseminated or metastatic breast
cancer is profoundly different than eradication of primary
localized breast disease. These forms of breast cancer are most
often chemotherapy resistant and nearly always fatal. Today there
is no satisfactory chemotherapy or any other therapy for these
cancers. In a marked departure from conventional theories and
methodologies, effective treatments for both ER.sup.+ and ER.sup.-
disseminated breast cancers have been devised.
[0049] Iron Metabolism and Disseminated Breast Cancer. It has been
found that diferric transferrin is unconditionally required for
both ER.sup.+ and ER.sup.- human breast cancer cell growth. It is
clear that ER.sup.+ cells are under both hormone and growth factor
control. However, ER.sup.- cancer cells no longer require growth
factors or hormones for proliferation (29,30). Only nutrients are
required. It is a common observation that hormone autonomous breast
cancer cells have also escaped the requirement for exogenous growth
factors. However, without an adequate supply of iron delivered by
transferrin, both ER.sup.+ and ER.sup.- breast cancer cells fail to
survive. Iron is required for DNA synthesis and other key metabolic
processes (38). Of the known types of chelators, the present
studies indicate that only those that remove iron from diferric
transferrin and serum ferritin will be useful for iron deprivation
in tumors. An example of a class of chelators that are able to
successfully withdraw iron from serum ferritin and diferric
transferrin are the .alpha.-ketohydroxypyridine chelators (144).
Other classes are also known, and some of these may also be
suitable for use as described herein for combined treatment
modalities. Deferoxamine does not remove iron that is already bound
to transferrin.
[0050] A second or alternative approach is to use monoclonal
antibodies against the transferrin receptor to prevent iron uptake.
This has yet to be developed into an effective treatment (42-44).
Monoclonal antibody therapies alone are often ineffective because
(i) there is a large supply of the competing natural ligand
available that competes with the antibody for receptor binding,
(ii) the natural ligands often have higher affinity for the
receptor than the blocking monoclonal antibody, (iii) antibodies
often do not escape the blood readily, and (iv) humanized
antibodies are required for repeated/prolonged treatments.
[0051] Another approach that can be employed as part of the breast
cancer eradication program is to block transferrin directly in the
plasma so that it will not be a source of iron for cancer cells.
Current results support the view that diferric transferrin can bind
to cellular transferrin receptors via either the "N" or "C" lobes
(118) or that binding is primarily via the "C" lobe (119). Because
the lobes have similar amino acid sequences, it is likely that the
same receptor recognition sequence is present in both. Most notably
however, the amino acid sequence of diferric transferrin that codes
for receptor binding is not known. This sequence will be identified
using techniques that are well known in the art (e.g. by phage
display technology) and specific monoclonal antibodies will be
raised to block transferrin binding to cellular receptors,
employing standard techniques. This will prevent the use of
serum-borne diferric transferrin by cancer cells and thus starve
cancer cells for iron. This approach does not suffer from several
of the problems of receptor binding monoclonal antibodies just
cited above. The anti-receptor recognition monoclonal antibody is
not required to leave the general circulation to be effective.
[0052] Iron Chelation Depletion and Combined Modalities. To deplete
iron from the diet, deferoxamine can be given intravenously or
other chelators used orally. This will further lower the blood
plasma content of diferric transferrin and increase the
effectiveness of the receptor sequence monoclonal antibody just
described above. This treatment should be combined with a low iron
diet. In addition, many other oral drugs are available to reduce
the effective body load of iron. These can be used in combination
therapies to deprive the cancer cells of necessary iron
sources.
[0053] Genetically and Chemically Modified Transferrin. In
addition, genetically and chemically modified transferrin can be
used to introduce lethal doses of specific toxins and cytolytic
enzymes that kill cancer cells. For example, RNAse can be
genetically engineered or chemically attached to transferrin, using
techniques that are well known in the art. Delivery of this
cytotoxic enzyme via the transferrin receptor can be expected to
cause cell death. Most importantly for this disclosure, the most
dangerous (i.e. rapidly growing and spreading) ER.sup.- breast
cancers over-express the transferrin receptor (129). Genetically
modified transferrin will be developed that is cytotoxic and/or
unable to act as iron donor to cells. This strategy focusing
directly on transferrin has the advantage of acting systemically
without regard to the issue of tissue penetration by receptor
blocking monoclonal antibodies or the necessity of developing
"humanized" monoclonal antibodies. In some instances it will be
preferred to combine modalities that interfere with iron metabolism
in order to achieve the most satisfactory and effective results.
The above-described rat mammary tumor models will be employed to
confirm the suitability of this treatment modality for human
trials.
[0054] Immunotherapy of ER.sup.+ Breast Cancer. As discussed above,
studies in cell culture with ER.sup.+ breast cancer cells have
shown that contact with IgA and IgM causes cell death within three
weeks (29,30). These results will be employed in immunotherapy for
breast cancer based on the smaller, more tissue penetrating, Fc
domains of polymeric IgA and IgM. The use of Fc fragments is
planned because of data indicating their importance in causing cell
growth inhibition. The rat mammary tumor models described above
will be used to continue testing in vivo.
[0055] The natural forms of IgA and IgM, as well as the Fc
fragments, can be administered intravenously. Several
immunoglobulins including IgA and IgM are already FDA approved for
human use. Those preparations, as well as other secretory
immunoglobulin preparations, are expected to be useful for
inhibiting cancer cell growth when administered in various
pharmacologic amounts. A suitable pharmaceutical composition must
provide IgA and/or IgM in a form that is capable of producing the
above-described inhibitory effect on estrogen dependent breast
cancer cell growth, and the immunoglobulin component must be able
to bind to the poly-Ig (Fc) receptor or poly-Ig-like (Fc) receptor
that mediates such inhibition. The preferred active compositions
(i.e., cell growth inhibitory) contain dimeric/polymeric IgA and
pentameric IgM and an activity-stabilizing medium (e.g., a steroid
hormone stripped non-heat inactivated serum or purified
compositions containing calcium), as described in U.S. patent
application Ser. Nos. 09/852,958 and 09/852,547 (PCT Published
Application Nos. WO 01/86307 and WO 01/85210), incorporated herein
by reference. To treat localized breast cancer, oral immunization
challenge will also be used to increase the number and function of
IgA and IgM secreting B immunocytes in the breast tissue and
thereby provide more inhibitory/killing immunoglobulins. This
treatment has the advantage of not requiring a clinical setting for
administration and being applicable to all women regardless of age
or physical condition. This immunotherapy can be combined with
tamoxifen anti-estrogen or combined with other immune modulating
drugs that increase the function of the secretory immune system in
the breast. Preventative and risk reduction methods and
compositions are described in co-pending U.S. patent application
Ser. No.______ (Atty. Dkt. No. 1944-01301) entitled "Anti-estrogen
and Immune Modulator Combinations for Treating Breast Cancer," the
disclosure of which is incorporated herein by reference. This
treatment mode may have special application to breast cancer in
situ, a form of the disease that has not left the ducts and often
recurs in the same breast after lumpectomy or in the other
breast.
[0056] Gene Therapy and Anti-estrogen Therapy Combined for ER.sup.+
Breast Cancer. A new function for the well-known anti-estrogen
tamoxifen has been discovered (29,30). Tamoxifen mimics the
estrogen reversible inhibitory properties of immunoglobulins IgA
and IgM to inhibit ER.sup.+ breast cancer cell growth. Tamoxifen
acts in the complete absence of estrogens. This indicates a
function unrelated to its classical interaction with the ER.sup.-
(79). In related studies (29,30) it has been recognized that the
cell surface receptor mediating the growth inhibitory effects of
IgA and IgM is a member of the immunoglobulin superfamily of
receptors and shares the properties of the poly-Ig receptor and Fc
family receptors. The receptor properties have been described and
discussed (29,30). Data indicate that tamoxifen only inhibits the
growth of cells containing this immunoglobulin receptor.
Furthermore, the immunoglobulin mediating receptor appears to be
under the control of either sex steroid hormones or thyroid
hormones. Hence, loss of the sex steroid hormone receptor (or the
appropriate thyroid hormone receptor from among several thyroid
hormone receptors) causes an equal loss of the superfamily
receptor. This information can be combined to develop a new, more
effective gene therapy for breast cancer. ER.sup.- cells lacking
the superfamily receptor can be restored to immunoglobulin control
by infection with an immunoglobulin binding receptor DNA bearing
virus vector, using conventional DNA transfection techniques. As a
result, tamoxifen will then become effective even with the
transfected ER.sup.- breast cancer cells, and can be used to kill
these cancers as is done today with ER.sup.+ cancers. This is an
entirely new combination of gene therapy and anti-estrogen
therapy.
[0057] Gene Therapy and Immunotherapy Combined for ER.sup.- Breast
Cancer. Another approach to eradication of ER.sup.- disseminated
breast cancer has been devised. Gene therapy can be targeted to
human breast cancers via the over-expressed transferrin receptor.
Gene therapy using the poly-Ig-like Fc receptor will be used to
reestablish immune negative control of autonomous breast cancers
that would otherwise grow uncontrolled. In related studies (29, 30)
it has been established that exposure of breast cancer cells
bearing this receptor to IgA or IgM leads to cell death by
arresting cancer cell growth. By developing gene therapy and
coupling it with either oral "immunization" to boost systemic
immunoglobulins or direct intravenous use of suitable human
immunoglobulins, new treatments for disseminated breast cancer can
be established. The rat mammary tumor models described above will
also be used to establish the first applications in vivo. Use of
athymic nude mice and human breast cancer xenografts will be used
to establish human relevance.
[0058] Disseminated or Metastatic Breast Cancer Eradication using
Radioactive .sup.59Fe. The radioisotope .sup.59Fe is a
high-energy.gamma. emitter that disrupts breast cancer cells in
culture by fragmenting DNA. This proposal plans the use of
transferrin as a .sup.59Fe delivery system to both disseminated
ER.sup.+ and ER.sup.- breast cancers. Because these cancers have
such a high requirement for iron bound to transferrin, the delivery
system may only need low concentrations of this high-energy
isotope. Rat mammary tumor models are available to investigate this
therapy, as described above. It is expected to be effective because
growing cells concentrate more iron than static normal cells. This
modality may be especially effective with blood replacement
therapy, described as follows.
[0059] Blood Replacement Therapy and Cancer Eradication. A number
of companies are now well advanced in the development of
"artificial blood". The FDA expects that within a year or two a
blood substitute will become available. Many treatments, such as
gene therapy and the above-described iron therapies may be
enhanced. Others such as standard adjuvant chemotherapy are
expected to be more selective and effective in substitute blood
where the levels of interfering substances can be regulated. This
approach may also increase the effectiveness of tamoxifen and the
newer "pure" antiestrogens. Since each treatment round is expected
to last only a few weeks to a month, a substitute blood product may
be employed in one of the above-described therapies, as part of the
present breast cancer eradication program.
[0060] This approach may have special application to the treatment
of disseminated breast and other cancers because artificial blood
can be prepared free of diferric transferrin, which is usually
present in physiological human blood at about 1.0 gram per Liter.
Addition of low concentrations of radiolabled diferric transferrin
to artificial blood will avoid the dilution caused by the natural
blood diferric transferrin and therefore increase the effective
dose of radioisotope to cancer cells without exposing the body to
high levels of radiation.
Part III: Reduction of Risk of Developing Breast Cancer
[0061] Overview. The risk of developing breast cancer for women in
the United States has been rising steadily for the past several
decades. It will soon approach one in eight. It is fortunate that
new treatments and more effective screening methods have kept
mortality rates from rising dramatically. Nonetheless, more than
one hundred women are lost per day to breast cancer in the United
States. Researchers and health care providers understand that the
first line of defense against this disease is prevention. If the
long term outlook for all women is to be improved, and especially
if our daughters to be free of this threat, the focus must now be
on finding a prevention.
[0062] As described above, a recent breakthrough in understanding
how breast cancer grows reveals that, in its initial stages, breast
cancer cells are inhibited and even killed by the secretory immune
system (29,30). That means a part of our immune system can stop
early cancer cells from growing. During adult life, breasts produce
milk or milk-like fluids. These fluids contain high concentrations
of three immunoglobulins, IgA, IgM and IgG1. These are passed from
mother to child during breast-feeding, and protect the child from
bacterial infections. As a result of this discovery, and
considering the fact that long duration breast-feeding is known to
reduce the risk of breast cancer, it is proposed that these
immunoglobulins are likely to protect the mother against breast
cancer via this newly discovered inhibitory mechanism. This
presents an unexpected opportunity to rethink the problem of
prevention and to apply new, unconventional approaches.
[0063] The present plans for a new type of oral immunization for
breast cancer were advanced by another remarkable fact about the
secretory immune system. The immunocytes of this system permit
mothers to protect their suckling offspring from infectious
pathogens. Because both mother and young child are exposed to the
same infectious agents at the same time, but only the mother can
develop immunity (or already has it), how does the young offspring
fight off disease? The answer is that infectious agents orally
entering the mother's body cause an antibody response in areas of
the intestinal mucosa called Peyer's patches. These lymphoid
structures cause the production of B immunocytes that ultimately
populate breast tissue. Once there, the B cells secrete milk-borne
immunoglobulins with specificity against the offending infectious
agents.
[0064] It is proposed that oral "immunization" is expected to be
most effective during a first susceptibility age range "window"
(e.g., during puberty, or 9 and 19 years of age), and/or during a
second "window," after menopause, when secretory immune system
function decreases sharply. If the secretory immune system of the
breast is stimulated at times when women are known to be most
susceptible to breast cancer, it might be prevented or at least the
risk of occurrence considerably reduced.
[0065] Finally, recent developments in mucosal cancers other than
breast cancer suggest another application of this discovery. As
discussed above, it is now understood that at least one mucosal
cancer is of bacterial origin. The bacterium Helicobacter pylori is
a Class I carcinogen thought to cause gastric cancer. This fact,
coupled with the discovery of new secretory immune functions in the
breast, supports the proposal that breast cancer might arise from
an infectious agent. Plainly, there is no known cause for 70% or
possibly more of breast cancers, although environmental carcinogens
are most often named as culprits. Nonetheless, bacterial
participation or a bacterial origin remains entirely possible. If
such was proven, the development of pathogen specific breast
immunity via oral challenge would offer a unique approach to
immunization.
[0066] In the history of cancer research, oral immunization has
been investigated mostly from the point of view of treatment.
However, there has been no serious application to either the
treatment or prevention of breast cancer, which is surprising since
oral immunization is so readily adaptable to mass populations of
women of all ages and all circumstances. Applying oral immunization
as a new means of preventing breast cancer is a preferred part of
the breast cancer total eradication program.
Oral Immunization to Reduce the Risk of Breast Cancer
[0067] During adult life, breasts produce milk or milk-like fluids
that contain high levels of three immunoglobulins, IgA, IgM and
IgG1. In cell culture these immunoglobulins not only block early
breast cancer cell growth, but if elevated for a period of time,
will kill breast cancer cells. This discovery presents the
unexpected opportunity for a new oral immunization for breast
cancer, taking advantage of a basic function of the secretory
immune system: the natural production of IgA and IgM.
[0068] It is proposed that oral immunization be administered when
it might be most effective, i.e., during the same susceptibility
age ranges that were identified in data collected from survivors of
the atomic bomb blasts during World War II, which showed an
unexpected pattern of breast cancer development. Those exposed to
the radiation between 9 and 19 years of age developed breast cancer
at much higher rates than survivors who were 30 years or older
(45). This meant there was a "window" early in life when breast
tissue was highly susceptible to DNA modifying mutations. This same
pattern became clear again when women survivors of Hodgkin's
disease were studied. Treatment of Hodgkin's requires agents that
cause DNA damage. Women treated between 11 and 19 develop breast
cancer at higher rates than women whose treatment began at later
ages (46). Again, young women appeared to have a "window" of
susceptibility. This same "window" is well known in experimental
animals exposed to carcinogens (47,48).
[0069] Young women might not be alone in experiencing increased
susceptibility, however, as secretory immune system function
decreases sharply after menopause. This coincides with the time
when breast cancer rates achieve near their highest levels (145),
and may represent a second "window" not previously recognized. If
the secretory immune system of the breast is stimulated at times
when women are known to be most susceptible to breast cancer, it
might be prevented or at least the risk of occurrence considerably
reduced.
[0070] Rat mammary tumor models are being used to define the
conditions for increasing breast tissue IgA/IgM secreting
immunocytes and determining if this protects against the DNA
synthesis dependent damage effects of estrogens as carcinogens (49)
as well as the effects of environmental carcinogens (36).
Conditions and appropriate oral bacterial challenges (i.e.
non-pathogenic and pathogenic E. coli) to increase breast cell B
immunocytes and therefore increased secretion of IgA/IgM are being
defined. It is known that women from the non-Western world have
high levels of antibodies to fecal E. coli in their milk. An
example study compares women from the United Kingdom and Sri Lanka
(146,148). These women also have the lowest risks of any worldwide.
In the present disclosure, it is now proposed that these
microorganisms, live or attenuated, or fragments or molecules from
them, will be potential agents for inducing breast cancer immunity
(153). Oral immunization is not the only administration route to be
considered. Nasal, rectal and vaginal administration must also be
considered (150,154). Antigen challenge may be required on a
routine schedule since oral immunity may not be permanent. Multiple
challenges are most likely necessary to maintain full immunity.
This means placing the challenge in a delivery form suitable to the
site of challenge. For orally administered treatments a tablet, a
food product, a food drink, or the like may be most useful and
readily accepted by women. When increased secretory immune function
has been established, the effect of the immunoglobulin cell-growth
inhibitors on attenuation of pre-malignant changes in breast ductal
cells will be evaluated as described (56-58).
[0071] Another route for presenting the bacterial antigens to women
of all ages on a routine schedule is in genetically engineered
food. Potatoes, tomatoes and bananas can be genetically engineered
to express foreign antigens such as those from a virus or bacteria
(151,152). Oral immunity expressed by mucosal tissues can then be
achieved on a routine basis by consumption of the generically
developed food product (151-155). This route for administration has
worldwide applications. This technology has not been previously
applied to immunization of breast, prostate, colon, kidney, ovary
or endometrial cancer.
PART IV: ERADICATION BY CONVENTIONAL IMMUNIZATION AGAINST THE
INFECTIOUS AGENTS THAT CAUSE BREAST CANCER
Infectious Origin of Breast Cancer and Mass Immunization
[0072] A bacterial origin of cancer is a growing theme, however
this has not always been the case. The work of Marshall &
Warren in 1982-1984 was a milestone, in which their identification
of a genus Campylobacter organism in gastric ulcers from biopsy
specimens changed our thinking completely. Notably, H. pylori was
unheard of 20 years ago. The notion that gastric ulcers were caused
by a bacterium was unprecedented. Initially, the skeptical
scientific community rejected this idea. Today it is a recognized
etiology of a major human disease (15).
[0073] Finding a clue to the origin of breast cancer comes from the
fact that 75% of breast cancer is invasive ductal carcinoma, it is
now proposed that the agent causing ductal carcinoma exists in the
ducts. As with all mucosal tissues, breast ducts are open to the
exterior; it is likely that infectious agents enter the ducts. The
question is can these organisms cause breast cancer?
[0074] Today it is recognized that bacteria are long-term
participants in cancer development (14,18,19,103) including gastric
(15,16), colon (50,51), cervical (17), and very likely prostate
(104) cancer. The concept that bacteria are involved in colon
cancer is more applicable to breast cancer than the H. pylori
model. Investigators studying the colon model offer some important
insights. The bacteria involved in colon cancer are not obvious at
histologic examination and do not cause ulcers or severe
inflammation as does H. pylori. Instead, Bacteroides in colon
produce fecapentaenes that are potent mitogens (i.e. they cause
cells to grow and divide) (51). By way of comparison, breast
atypical hyperplasia (i.e. increased growth rates above normal) is
known to be a pre-cancerous condition that also does not show
severe ulceration or major inflammation. It is expected that breast
bacteria also cause DNA synthesis and cell growth necessary to make
cancer-causing mutations permanent.
[0075] Human milk contains many microorganisms (52-55). They are
believed to be skin and nipple contamination of expressed breast
milk. Then again, in some samples, the organisms were not usual
skin flora (52). In one larger study, several pathogenic organisms
were found along with non-pathogens (53). No virus particles were
identified in the milk. The fact that bacteria are present on the
nipple and surrounding skin certainly leaves open the possibility
that they might migrate into the ducts. To date, no microbes have
been investigated from the perspective of causative agents of
breast cancer.
[0076] Data supporting the likely presence of at least low levels
of bacteria in ducts comes from two different sources. First, human
milk lipids have been modified by chemical reactions to generate
genotoxic agents (56-58). The presence of these agents is measured
directly by DNA damage of breast cells obtained from milk. Because
genotoxic components are in freshly expressed milk, the products
are thought to be endogenous to the breast. In another study,
samples of freshly expressed human milk contained
N-nitroso-dimethylamine, nitrate and nitrate reducing
microorganisms (59). This study concluded that the compounds arose
endogenously. These reactive nitrogen compounds are likely cancer
causing agents in other tissues. It highly probable that reactive
lipids and nitrogen compounds are formed endogenously in human milk
ducts.
[0077] Considering that the development of breast cancer is a
multi-step process that often leads to tumor cell heterogeneity
(111), it is now proposed that the steps leading from normal duct
epithelium to hyperplasia to pre-malignant changes and finally to
in situ carcinoma and invasive ductal carcinoma (112) are caused by
a relatively continuous source of mutagenic agents that are present
over a number of years. Furthermore, because each change at each
step requires DNA replication in order to become permanent, it is
likely that both estrogens and the causative bacteria induce cell
proliferation. Since bacterially induced mammalian cell
proliferation related to cancer development is known, the proposed
model for bacterial causation takes into account the very well
known (112) progression of normal breast epithelium as it
transitions to invasive ductal carcinoma (i.e. the form in 75% of
breast cancers).
[0078] In further studies, human milk, breast cancer samples and
normal breast tissue (reduction mammoplasty) will be examined for
bacterial content. They will be analyzed for bacterial content by
culture (aerobic and anaerobic) and PCR methods (62-65). Example
techniques to be applied include (i) use of specific PCR primers
for known and new bacteria, (ii) PCR amplification of conserved 16S
rRNA sequences, and (iii) RDA-PCR which is also called "reverse
PCR". These can be used to identify unique infectious agents in
tissues, even in paraffin embedded specimens (61). PCR technology
pertinent to the identification of most microbes that this study
might encounter is now being applied.
[0079] Next, colony-derived bacteria will be used in the "Ames
test" to identify mutagen production (66). Culture medium from the
bacteria isolated can be tested directly for mutagenic activity
using any of several strains of Salmonella. Candidate organisms can
be grown plus and minus human milk components to determine the
source of the mutagenic agents. The different types of existing
screening methods have been reviewed (67). Improvements in the Ames
test have been introduced to provide more quantitative evidence
that the assay is providing significant results with respect to
cancer bioassays (68,69). The results of this test will establish
which bacterial isolates produce mutagenic metabolites. The Ames
test can also be applied to demonstrate that the bacteria cause an
"oxidative burst" mediated by neurophils and macrophages. In this
case, the leukocytes are incubated with the bacteria to generate
the active mutagenic species. This approach resolves the issue of
whether the products of the bacteria are the mutagens themselves or
if the activation of leukocytes is required.
[0080] Bacteria that meet the criteria described above will be
cultured and the medium tested with non-tumorigenic human breast
epithelial cells (Clonetics, San Diego, Calif.) or epithelial cells
derived from human milk (56,58) for transformation activity. The
human milk derived HBL-100 non-tumorigenic cells are also
candidates for this assay. The cells will be tested for colony
formation in soft agar. Tumor or transformed cells will form
colonies. There is a strong correlation between colony formation in
soft agar and tumorgenicity in animals. This approach will confirm
that the Ames test translates to transformation of human breast
cancer cells. Candidate cell lines will be analyzed for tumor
formation in athymic nude mice.
[0081] In addition to the above analyses, the bacterial isolates
are expected to have an additional immunoprotective mechanism.
Breast secretions contain high concentrations of secretory IgA that
kill bacteria by the known antigen-antibody recognition function.
This is a first line of protection against breast duct infections
by many strains of bacteria. However, some bacteria can escape IgA
killing by secreting proteases that cleave the IgA into inactive
Fab and Fc fragments.
[0082] In a final test, an animal model will be sought to determine
if infection leads to breast cancer development. Several strains of
inbred and outbred rats are highly susceptible to breast cancer
induction. Candidate bacteria will be introduced into the breast by
milk pump (60) and tumorigenesis monitored.
[0083] The final step will be to use the attenuated organism, or
entities derived from the organism, to test for oral induced
immunity in human breast milk as has been described (148,149). Once
immunity against the mutagenic bacteria is established in human
milk, studies can move forward to determine if this method reduces
the risk of developing breast cancer.
[0084] Surrogate end points will be used to estimate the
effectiveness of oral route administration of immunogen. Because a
full clinical trial of an oral immunization may require five or
more years to establish efficacy, DNA changes in cells isolated
from milk or breast fluids of women being administered the
treatment will be studied. By using this surrogate end point
approach, a reduction in genotoxic (i.e. mutagenic) events can be
identified within months. This will provide data to support more
expensive long-term clinical studies.
[0085] The same antigen preparation protocols can be used with
direct immunizations by standard methods such as intramuscular
injection. Many approaches to standard immunizations are known and
commonly employed in worldwide programs to eradicate infectious
diseases. The oral route of immunization utilizing the secretory
immune system is considered preferable because the antibody is
delivered directly into the intraductal space where the causative
bacteria are then neutralized.
Conclusions
[0086] The program and procedures described above advance directly
to the core problem of eradication of breast cancer. Today there
are women battling breast cancer and others already developing the
pre-malignant changes leading to the disease. For them eradication
means availability of effective treatments, especially for
chemotherapy resistant metastatic cancer. Treatment will likely
remain an important issue for several years to come, and must
therefore be given serious continuing consideration. Very
definitely, this means developing new methods of eradicating
metastatic breast cancer. One aspect of the present eradication
program departs from the usual chemotherapy regimens to exploit the
nutritional requirements for growth of breast cancer. Cancer cells
grown in culture invariably require iron in the form of diferric
transferrin for growth. With the advent of the very powerful tool
of serum-free defined cell culture (113-118), this requirement
could be established conclusively (29,30,113-117,135). Variations
in this requirement usually result from differences in the level of
storage of iron as ferritin (126) in the cells. When ferritin
stores are depleted, iron must be acquired from the outside of the
cells via the specific iron carrier transferrin (136) and
internalization via specific cell surface transferrin receptors
(127,128,134). The same logic/hypothesis employed in the breast
cancer eradication program is extendable to all epithelial cancers
(80% of the total cancers in humans) and for lymphoid origin
cancers, sarcomas (i.e. cancer of the connective tissues and
muscle) and cancers of the bone and nervous system. Indeed, it is
absolutely clear that iron is required for the growth of all cells
because of its involvement in metabolic processes and DNA synthesis
(124,130,131). It can be readily appreciated that the disclosed
concepts and procedures involving the exploitation and/or
interruption of iron metabolism required for cancer cell growth
apply to many if not most cancers of humans.
[0087] The eradication of breast cancer will likely come from a
different direction than the common themes of genes and cell
signaling pursued by so many investigators today. The work leading
to the new eradication program comes from the discovery of the role
of the secretory immune system in estrogen responsive breast cancer
cell growth, described in the two U.S. patent applications cited
above. Based on this discovery, the plan for an "oral immunization"
to protect breast cells from DNA synthesis dependent mutagenic
events has been devised. The goal is to reduce risk from the
current 1 in 8 to a level of 1 in 40 or 50. This plan is consistent
with the growing, but still widely ignored, concept that the
population of the Western world is placed at higher risk for
mucosal cancers because we are "too clean." Our immune systems are
not challenged sufficiently to protect us.
[0088] An additional immune based eradication plan finds support in
an unexpected source. It is known that bacterium Helicobacter
pylori is the most common cause of gastric ulcers. It also is the
first bacterium to be a definite cause (Class I) of a human cancer
as rated by the International Agency for Cancer Research (14,15). A
number of infectious agents have been documented to cause or
contribute to human cancers (14-19). However, this information has
had little or no impact on the search for the origin of breast
cancer. More commonly, it has been proposed that environmental
chemicals cause the possible majority of human cancers. Indeed,
despite advances in defining various breast cancer risks due to
chemical exposure (20), the perplexing element of randomness
without clear indications of chemical exposure has not been
explained. An infectious origin is random, and therefore a
reasonable alternative. Breast cancer risk is sometimes familial
but most often is not genetic (i.e. inherited) for most women.
Indeed, this is a characteristic of the incidence pattern of ulcers
in H. pylori infected families (16). While not all infected
cohabitants develop gastric ulcers or gastric cancer, members of a
family tend to have higher incidences than average. By seeking
infectious agents as either the cause or as major contributors to
breast cancer development, and then using that information to
develop appropriate immunizations, it is expected that the
incidence of breast cancer will be reduced or eliminated.
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[0251] While the preferred embodiments of the invention have been
shown and described, modifications thereof can be made by one
skilled in the art without departing from the spirit and teachings
of the invention. The embodiments described herein are exemplary
only, and are not intended to be limiting. Many variations and
modifications of the invention disclosed herein are possible and
are within the scope of the invention. For example, the foregoing
discussion specifically focuses on the eradication of breast
cancer, however the same or similar approaches can be employed to
eradicate other types of cancers of mucosal tissues, including
prostate, ovary, endometrium, cervix, vagina, colon, kidney, lung
and nasopharynx. Cancers of those tissues, together with breast
cancer, account for 80% of all human cancer. The disclosures of all
patents, patent applications and publications cited above are
hereby incorporated herein by reference. The discussion of certain
references in the Description of Related Art, above, is not an
admission that they are prior art to the present invention,
especially any references that may have a publication date after
the priority date of this application.
* * * * *