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Tel Hashomer Medical Research, Infrastructure and Services Ltd.
Tel: +972-3-5305998 Fax: +972-3-5305944
Tel Hashomer Medical Research
Infrastructure and Services Ltd.
Novel Therapeutics for Human Diseases
Contact :
Sylvie Luria PhD.
Technology Transfer and Business & Development Manager
Tel Hashomer Medical Research, Infrastructure and Services Ltd.
Tel: +972-3-5305998 Fax: +972-3-5305944 Cell: 052-6667277
[email protected]
http://research.sheba.co.il/e/
1
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Cancer Therapy Targeting the SIL Regulator Gene
The STIL (SIL) gene, SCL (Stem Cell Leukemia TAL1 Interrupting Locus), located on
chromosome 1, was cloned from the most common chromosomal rearrangement in T cell
Acute Lymphoblastic Leukemia. It encodes a 150 kDa cytosolic and centrosomal protein
conserved in vertebrates. STIL is tightly regulated during the cell cycle. It is expressed only
in proliferating cells, reaching a peak in mitosis during which the protein is phosphorylated
and then degraded upon entrance to the next cell cycle. The critical requirement of STIL for
cell growth, proliferation and survival during embryonic development, and its regulation
during the cell cycle suggest that STIL might have a role in tumorogenesis.
We have demonstrated increased expression of STIL RNA and protein in a wide variety of
cancers. Further analysis revealed that genes with expression patterns similar to STIL were
mitotic checkpoint genes regulated by the transcription factor E2F1 known to be activated in
proliferating cells. The mitotic index is increased with expression of STIL and its
expression in epithelial cancers is associated with metastases and poor prognosis.
Inhibition of STIL expression by a specific anti STIL siRNA in colon cancer cells was
established and demonstrated - STIL blocks mitotic entry and causes apoptosis of this
colon cancer cells in-vitro and in-vivo. Similar results have been obtain on a variety of
cancer cells representing the most common types of human cancer (Cervical, lung, breast,
prostate, gliomas and renal cancer) using anti STIL siRNA oligonucleotides. The lethal
effect of the knockdown of the human SIL is completely rescued by transfection of the
murine STIL, demonstrating the specificity of the siRNA mediated approach. STIL is not
necessary, however, for survival of all normal proliferating cells as we have mouse
embryonic stem cells and fibroblasts that are negative for the STIL gene.
We conclude that STIL is a good target for treating abnormal proliferation and cancer.
Anti STIL activity was demonstrated to be effective in inhibiting tumor cell development in
vitro and in vivo models for variety of human cancer models.
2
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Methods of Predicting Clinical Course and Treating Multiple Sclerosis
Multiple sclerosis (MS) is the most common demyelinating disease of the central nervous
system (CNS) affecting young adults (disease onset between 20 to 40 years of age) and is
the third leading cause for disability after trauma and rheumatic diseases, with an estimated
annual cost 34,000 USD per patient (total life time cost of 2.2 million USD per patient).
The disease is characterized by destruction of myelin, associated with death of
oligodendrocytes and axonal loss. The main pathologic finding in MS is the presence of
infiltrating mononuclear cells, predominantly T lymphocytes and macrophages, which
surpass the blood brain barrier and induce an active inflammation within the brain and spinal
cord. The neurological symptoms that characterize MS include complete or partial vision
loss, diplopia, sensory symptoms, motor weakness that can worsen to complete paralysis,
bladder dysfunction and cognitive deficits, which eventually may lead to a significant
disability. The associated multiple inflammatory foci lead to myelin destruction, plaques of
demyelination, gliosis and axonal loss within the brain and spinal cord and are the reasons
contribute to the clinical manifestations of neurological disability.
The etiology of MS is not fully understood. The disease develops in genetically predisposed
subjects exposed to yet undefined environmental factors and the pathogenesis involves
autoimmune mechanisms associated with autoreactive T cells against myelin antigens. It is
well established that not one dominant gene determines genetic susceptibility to develop
MS, but rather many genes, each with different influence, are involved.
We have discovered a unique gene set for Prediction and Diagnosis of Benign Multiple
Sclerosis – New target for MS drug .
Using high throughput gene expression analysis we have identified and characterized one of
the major junctions involved in the determination of disease course amongst MS patients.
Furthermore, we used the promoters and transcripts of these genes to generate reporter cell
lines for high throughput drug discovery system. The promoters and transcripts of genes that
participate in a certain pathway can serve as reporters for cellular drug discovery screening
system for that particular pathway. The cell based functional drug discovery system based
on analysis the signature map in human cells before and after drug treatment have two
laboratory applications: 1) Primary cultured PBMS from MS patients or test cell culture
treated with variety of drug candidates followed by multiplex assay to analyze expression of
related genes: 2) Mammalian cells stably transfected with plasmids containing the promoters
of BMS-specific genes coupled to variety of reporter genes for detection and drug
discovery. Both assays combined with simultaneous multiplex analysis of wide range of MS
related inflammatory related proteins such as cytokines, adhesion molecules and nerodegeneration factors in cultured media. Assay enables the discovery of potential drug to treat
MS and variety of Inflammatory diseases.
3
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Novel Therapy for anti-phospholipid syndrome - APS
Anti-phospholipid syndrome (APS) is an antibody-mediated autoimmune disease. APS, is a
life threatening blood-clotting disorder, also known as Hughes’ syndrome, is characterized
by the presence of anti-phospholipid (aPL) beta-2-glycoprotein-I (2GPI)–dependent
antibodies or lupus anticoagulant associated with thromboembolic phenomena,
thrombocytopenia, recurrent fetal loss, CNS, heart and other organs involvement. The
diverse clinical manifestations are associated with elevated titers of circulating anti-beta-2glycoprotein-I (b2GPI ) antibodies (Abs), and lupus anticoagulant. This autoimmune disease
afflicts up to two million patients, including many lupus patients, in the United States and
Europe. Unlike the typical stroke patient, these patients often experience their first stroke,
heart attack or miscarriage in their 20s and 30s and have twice the probability of a
recurrence.
We have identified unique peptides corresponding to 2GPI that were able to prevent anti2GPI related endothelial cell activation in-vitro and induction of experimental APS in an
animal model. Antibodies specific to these synthetic peptides, induced by common bacteria
and passively transferred into naïve mice, induced experimental APS. Previously, our
studies were based on neutralization of anti-2GPI Ab function by specific synthetic
peptides. And recently we have developed a methodology that addresses specific clone
death induce apoptosis of the B cells which express the anti-2GPI immunoglobulin or the
mature B cell population which secret the pathogenic autoantibody. There results were
further analyzed in -2GPI in-vitro system and in experimental APS animal model. In
addition,
The results suggest a novel therapeutic approach in which B cells that will secrete
pathogenic antibodies are targeted specifically. This strategy might also be applicable in the
treatment of other diseases that are humorally mediated by antigen-specific antigen
4
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Rejuvenation of Myocardial Scarring
The human heart has little or no capacity to regenerate. This is a major medical problem,
because inadequate regeneration contributes to myocardial scarring, heart failure,
arrhythmia, and death. As a single cause of death, ischemic heart disease is one of the
leading causes of mortality worldwide among people ages. This disease results from
occlusion of cardiac vessels leading to loss of cardiomyocytes causing million deaths every
year.
Scar thinning, enlargement and spherical deformation of the left ventricle with a
concomitant increase in wall stress are key elements in the pathogenesis of adverse
remodeling and heart failure after myocardial infarction (MI). It is suggested that left
ventricular (LV) remodeling may contribute independently to the progression of heart
failure and death. Thus, any intervention that will increase scar thickness and strength will
reduce LV remodeling.
Surgically, cardiac transplantation is a potential treatment for many patients. However, lack
of donors combined with an increasing number of patients has led to the search for other
surgical strategies. Patients with symptomatic large left ventricular aneurysms have been
treated with resection of the aneurysm and closure of the left ventricle either directly or by
implantation of a patch. Scar areas of the left ventricle have also been successfully treated
by the latter method. According to the law of Laplace, large dilated ventricles have
increased wall tension and thus increased oxygen consumption. Based on this fact, Batista
and coworkers have reduced the volume of enlarged left ventricles in patients in terminal
heart failure by removing a wedge of myocardium from the apex of the heart towards the
base of the left ventricular free wall. This method is currently not recommended for
treatment of heart failure because of high surgical failure rates.
After MI, failure of extracellular (ECM) support has been associated with LV wall thinning
and slippage of myocyte fascicles. This adverse remodeling process has been termed "infarct
expansion" and occurs in the absence of additional myocyte injury or alterations in LV
loading conditions. This post-MI remodeling process is a clinically significant problem in
that it can lead to LV dilation and dysfunction, the progression to heart failure. Indeed, this
post-MI remodeling process, which includes changes in ECM structure and composition, is
an independent predictor of morbidity and mortality. It has been postulated that an
acceleration of ECM degradation occurs within the scar and the myocardium surrounding
the MI (border zone) and facilitates the infarct expansion process in this later phase of postMI remodeling.
We have developed a unique approach to rejuvenate and strength an old scar after MI. The
rational for this is that collagen matrix play a central role in the remodeling of the infarct
zone and the ultimate transition to cardiac decompensation. Our experiments suggest the
stimulation of new collagen and elastic fibers growth remodeling of the failing myocardium.
Our preliminary results also suggest that this approach is safe and might recruit cardiac stem
cells to the site having a functional synergistic positive effect.
5
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Elimination of Clonal Responses to Biological therapeutics
Biological drugs have opened new therapeutic horizons for treating human diseases, but
have also brought with them issues related to immunogenicity, long-term efficacy, safety
and cost. During treatment with biopharmaceutical proteins, including insulin, growth
hormone, granulocyte-macrophage colony-stimulating factor, factor VIII, erythropoietin,
antibodies and interferons, there is an immune response to the biopharmaceutical agent,
including the generation of anti-drug antibodies (ADAs) and hypersensitivity reactions.
ADAs may reduce the clinical efficacy of these agents by blocking or neutralizing their
biologic activity and may have other biologic effects. The potential to induce ADAs after
treatment with biologics is a safety issue that has become an important consideration in the
development of biologics and a critical aspect of regulatory filings.
For example, antibodies develop in 20 % to 40% of patients with severe hemophilia treated
with human factor VIII; the presence of these antibodies can result in tolerance to the
clotting effects of this agent. Similarly, a proportion of patients treated with interferon alpha
develop antibodies, which inhibit its therapeutic effects. Therefore, it is important to test for
neutralizing antibodies during treatment with these agents, particularly in patients who are
unresponsive to treatment or have breakthrough disease.
Concurrent immunosuppressive drugs and pre-infusion steroids can decrease antibody
formation to biological drugs. We have discovered a composition and methods for reducing
undesired immune response to biological drugs and autoimmune antigens using unique
combination therapy. Our initial studies demonstrated their efficacy in several applications.
The global market for biopharmaceuticals, which is currently valued over US$ 50 billion, has been
growing at an impressive compound annual growth rate of 19% over the previous five years. With
over one third of all pipe-line products in active development are biopharmaceuticals, this segment is
set to continue outperforming the total pharmaceutical market and could easily reach US$100 billion
by the end of the decade.
6
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Novel Treatment for Neuro-developmental Disorders
Neurodevelopment disorders are now recognized to be the result of abnormalities in brain
development due to both genetic and environmental/biological causes. In total, these
conditions affect approximately 1-3% of the population, frequently resulting in substantial
economic, physical and emotional burdens to individuals, families, and society. Therefore,
research into the pathogenesis and treatment of these disorders, aimed at the cure and
amelioration of their effects on individuals, families, and society, is of considerable
importance.
Because of the brain disruption which is at the root of all neurodevelopmental disorders,
there are several common features across the variety of disorders in this group. Some of
these include difficulties with motor development, sensory integration difficulties, speech
and language delays and a range of cognitive difficulties including learning disabilities, poor
organizational skills, poor self regulation and behavioral difficulties.
We are studying the Rett syndrome, a genetic neurodevelopmental disorder affecting
mainly females. The genetic cause for most of the cases are mutations in the X-linked
MECP2 gene. Mecp2 protein is a transcription regulator of several known and still unknown
genes. One of the main affected genes is the BDNF gene .The level of BDNF is decreased in
MECP2 mutated and deficient mice. It was already shown that BDNF null mice have
clinical features which resemble the MECP2 deficient mice and over expression of BDNF in
MECP2 deficient mice causes delay and partial rescue of the clinical phenotype of these
mice. Copaxone is an immune modulator but lately one of his major effects was found to be
elevation of BDNF in EAE mice and in MS patients. It is a safe drug to use with very mild
side effects.
We have found a compound to be given to Rett syndrome girls and cause elevation of
BDNF protein expression in their brains, and potentially relief Rett symptoms.
Neurodevelopmental disorders are thought of as beginning before birth, and many such as
Down syndrome clearly do. Clinically, however, other such disorders may unfold over
months such as Mental Retardation, Autism, Rett Syndrome or years as Asperger
Syndrome, Fragile X and Dyslexia. These applications are candidate to be analyzed with
this compound.
7
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Novel Pharmaceutical composition to treat Breast Cancer.
Breast cancer is the most common cancer among women. Despite recent advances,
treatment of breast cancer is still unsatisfactory and the median survival of patients with
metastatic disease is about two years. Moreover, current chemotherapies used to treat breast
cancer are associated with severe side effects. The identification of novel and less toxic
therapies is of major importance. We have identified a peptide hormone GLP-1, which is
secreted from endocrine L cells, located in the distal ileum and colon. The hormone is
secreted in response to food intake, stimulates insulin secretion and sensitivity and inhibits
glucagon secretion. In addition, GLP-1 promotes pancreatic β-cell proliferation. GLP-1 also
appears to be a physiological regulator of appetite and food intake. These actions are
mediated mainly through GLP-1 receptor (GLP-1R), a seven transmembrane G-protein
coupled receptor. Due to these actions, GLP-1 or GLP-1 receptor are targets for the therapy
of type 2 diabetes. GLP-1 is a 31-amino acids long peptide, with a short half-life in the
circulation (less than 2 minutes), which is rapidly inactivated by the ubiquitous proteolytic
enzyme dipeptidyl peptidase-4 (DPP-4). Therefore, the use of GLP-1, and GLP-1R
analogues with a prolonged half life are currently being evaluated.
As both diabetes and obesity are associated with breast cancer, and as GLP-1 induces insulin
sensitivity, we studied the effects of the analogues on breast cancer cells.
Our results show that the analogues, which are already approved to treat type 2 diabetes,
are potent and selective inhibitors of breast cancer cell proliferation, and can enhance the
activity of doxorubicin.
8
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Novel Pharmaceutical composition in the treatment of pancreatic cancer.
Klotho is a transmembrane protein, which is expressed in the brain and kidneys, as well as
in the pancreas. Klotho can be shed and act as a hormone, and has been identified previously
as an inhibitor of the insulin growth factor (IGF)-1 pathway in hepatocytes and muscle cells.
The IGF-1 pathway is known to play a major role in the development of pancreatic cancer.
Pancreatic cancer is a fatal disease and the 5 years survival of patients who suffer from
advanced disease is less than 10%. Current treatment for metastatic pancreas cancer consists
of chemotherapy and is unsatisfactory, with a median survival following chemotherapy of
about 6 months.
We studied the effects of klotho on pancreatic cancer cell lines and identified, for the first
time, growth inhibitory activity of klotho on the pancreatic cancer cell lines PANC-1,
miapaca and Colo-357. Growth was inhibited by either over-expression of klotho or
treatment of the cells with soluble klotho, where soluble klotho inhibited PANC-1 cells
growth up to 50%. We also found additive effects of klotho and various chemotherapy
agents and noted more than 70% inhibition of PANC-1 cells by the combination of klotho
and low dose of chemotherapy.
Thus, our in vitro and in vivo data suggest klotho, for the first time, as a novel therapy for
pancreatic and breast cancer and indicate a possible additive effect for klotho when
administered with standard chemotherapy.
Our data suggest high levels of klotho protein expression in normal breast samples and
normal pancreatic cells, but very low expression in cancerous breast cells or cancerous
pancreatic cells. In addition, noted inhibition of breast cancer cell growth following overexpression of klotho protein, and growth enhancement of klotho protein-expressing cells
following klotho protein knock-down; and revealed modulation of the IGF-1 and the insulin
pathways by klotho protein. Taken together, the results suggest klotho protein as a novel
breast cancer tumor suppressor. Moreovere, we have found additive effects of klotho and
various chemotherapy agents and noted more than 70% inhibition of PANC-1 cells by the
combination of klotho and low dose of chemotherapy. Animal studies demonstrate a
therapeutic effect of Klotho. Treatment of mice with Klotho protein resulted in a significant
dose response inhibition of tumor growth. The present invention relates to the use of the
klotho protein for the treatment and diagnosis of cancer, such as breast cancer and
pancreatic cancer as well as other IGF-1 dependent cancers.
9
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New approach for gene silencing for treatment of psoriasis
microRNAs (miRNA), are a group of hairpin shaped gene transcripts whose function and
mechanism of action is being recently revealed. These molecules are transcribed in the
nucleus, transported to the cytoplasm, transformed by a series of enzymes to the active short
double stranded RNA duplexes which exert their suppressive effect on the translation of
multiple mRNAs. As compared to artificial short interfering RNAs (siRNA), miRNAs exert
their effects with less than full sequence homology, thus they are able to inhibit the
translation of multiple downstream genes, thus acting as master genes. Few hundreds of
such genes were discovered, and their essential role in the control of cell growth and
differentiation and their aberrant expression in acquired and genetic disorders is being
revealed. The therapeutic implication of miRNA discovery resides in the understanding of
their downstream targets and the possibility to predict siRNA sequences that will be
effective in interfering with the expression of the miRNA or it target. In addition, we are
developing a novel platform technology approach for gene silencing (psiRNA). The study
involves the synthesis of unique siRNAs conjugated to target promoter sequences of genes
that are essential for cell survival and/or proliferation.
We have developed a biological system for psoriasis, including animal models bearing
human psoriatic affected skin. We have discovered several miRNAs, differs significantly
between psoriatic uninvolved a normal skin for their expression. Analysis of the role and
downstream possible targets of these miRNAs showed the involvement in controlling the
expression of specific genes. Among those genes, bFGFR is associated with keratinocyte
hyperproliferation and was found to be overexpressed in psoriasis and IRAK2 which is part
of the IL-1 pathway that is strongly related to psoriasis. The platform psiRNA approace is
currently validated in cell models of cancer and psoriasis.
10
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FZD7 for targeting tumor stem cells
FZD7 is a member of the 'frizzled' gene family, encoding 7-transmembrane domain receptors for
Wnt signaling proteins. Binding of Wnt proteins to the FZD7 receptor results in activation of the
canonical Wnt pathway which usually leads to cell proliferation and survival. FZD7 is known to be
over expressed in several malignancies including Colon cancer, Esophageal cancer, Gastric cancer,
Wilm's tumor and more, compared to the normal tissue from which each tumor develops. It is
becoming clear that many, if not most, malignancies arise from a population of cells that exclusively
maintain the ability to self-renew and sustain the tumor via the expression of tumor-progenitor
genes. These ‘‘cancer stem cells’’ are often biologically distinct from the differentiated cancer cells
that comprise most of the tumor bulk. Since cancer stem cells are believed to be primarily
responsible for tumor initiation as well as resistance to chemo- and radiotherapy, their persistence
after such treatments may account for relapsing disease and metastasis in many malignancies.
Taking into account the Wnt pathway established role in enabling stem cell properties ('stemness') as
well as FZD7 over expression in multiple cancers in which the Wnt pathway is deranged, FZD7 is a
potential therapeutic target for numerous malignancies both as a biomarker for cancer stem cells and
as a target for attenuating Wnt signaling in these cells which in turn is likely to lead to cancer cell
death.
Wilms tumor (WT), a pediatric kidney cancer can serve as an excellent model for studying malignant
renal stem cells leading to tumor initiation and progression. An important link between aberrant Wnt
signaling and WT formation has been established. We have observed frizzled7 (FZD7) up regulation
in WT compared to normal kidney and showed FZD7 protein expression on a subset of WT cells.
Unexpectedly, incubation of WT cells with anti-FZD7 Ab resulted in apoptosis and cancer cell
death, suggesting possible manipulation of the tumor with antibody therapy. Further analysis of
additional WTs has identified tumours from which a viable FZD7(+) cell population can be isolated,
which enabled characterization of these cells as potential Wilm’s tumor stem cells. These novel
observations highlight the Wnt pathway and especially FZD7 as a therapeutic target in Wilm’s tumor
and possibly in additional FZD7 over-expressing cancers.
11
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Exendin-4 and related active pharmaceutical ingredients such as glucagonlike peptide-1 hormone, derivatives or analogues, in the treatment of cancer
Breast cancer is the most common cancer among women. Despite recent advances,
treatment of breast cancer is still unsatisfactory and the median survival of patients with
metastatic disease is about two years. Moreover, current chemotherapies used to treat breast
cancer are associated with severe side effects. Glucagon-like peptide 1 (GLP-1) is a
hormone secreted from endocrine L cells in the distal ileum and colon in response to food
intake. It stimulates insulin secretion and sensitivity and inhibits glucagon secretion. In
addition, GLP-1 promotes pancreatic β-cell proliferation and differentiation. These actions
are mediated mainly through GLP-1 receptor (GLP-1R), a seven-pass transmembrane Gprotein coupled receptor. Due to these actions, GLP-1 or GLP-1 receptor are targets for the
therapy of type 2 diabetes. GLP-1 is a 31-amino acids long peptide, with a very short halflife in the circulation, and it is rapidly inactivated by the proteolytic enzyme dipeptidyl
peptidase-4. Therefore, GLP-1 analogs with a prolonged half life were sought. One of these
analogs is exendin-4 (Ex4), which is isolated from Heloderma suspectum (Gila monster
lizard) venom and is a potent GLP-1R agonist. Ex4 has been approved by the FDA for type
2 diabetes treatment.
As both diabetes and obesity are associated with breast cancer, and as GLP-1 induces insulin
sensitivity and is a differentiation factor, we studied extensively the effects and mechanism
of action of Ex4 on breast cancer cells. Exendin-4 almost completely inhibited proliferation
of human breast cancer cells but not of human primary liver cells. Moreover, Ex4 enhanced
the activity of doxorubicin, a commonly used chemotherapy for breast cancer, on breast
cancer cells. Ex4 treatment induced elevation of the tumor suppressors p53 and p21 in breast
cancer cells, but down-regulated it in human primary liver cultures. Also, Ex4 treatment
increased apoptosis of breast cancer cells, as evidenced by increased PARP cleavage,
whereas it decreased apoptosis in primary liver cells.
Our detailed cellular and biochemical analysis and results indicate Ex4 is a potent and
selective inhibitor of breast cancer cell proliferation. Moreover, Ex4 can enhance the activity
of standard chemotherapy drugs.
12
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Anti-Microbial and Anti-Adherent polypeptides from Sea Species
Different aquatic species, such as sea anemones, are natural targets for microbial
colonization. To prevent that, they developed defensive polypeptides, which prevent the
adhesion of a wide range of biofilms. Biofilms consist of diverse microorganisms linked to
a surface by substances they secrete. Biofilms formed on edible plants, water supply and
medical equipment (especially catheters and other indwelling devices) are a major source of
infection and contamination. Bacteria in biofilms often outlive antibiotic treatment.
Our invention comprises a large set of proteins and polypeptides derived from sea species
which prevent bacterial adhesion. Such compounds provide innovative ways for the
prevention of microbial biofilm development on a variety of surfaces, such as orthopaedic
implants, stents, catheters. In addition they may be used for sanitation of food preparation
areas, plant protection againt yeast and moulds, and targeted therapy for preventing or
reducing biofilm formation in various human diseases such as infectious kidney stones,
cystitis, dental caries, chronic otitis media, bacterial endocarditis, osteomyelitis, wounds,
and acne. These polypeptides are resistant to organic solvents, lyophilization and high
ambient temperatures.
Currently, we have identified and characterised several proteins and peptides that tested for
their ability to prevent and reduce biofilm formation. The know-how includes the isolation
methods as well as methods for assessing the beneficial properties of the compounds
obtained.
We are developing several medical and cleantech applications using these
peptides.
13
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Melanoma Diagnosis and Treatment by CEACAM-1 Antibodies
Carcinoembryonic Antigen Cell Adhesion Molecule -1 (CEACAM1) is a transmembranal
glycoprotein that belongs to the carcinoembryonic antigen family and to the larger
immunoglobulin superfamily. CEACAM1 is expressed on various normal cells of epithelial
and endothelial origins, as well as on activated lymphocytes. Various functions are
attributed to CEACAM1, including regulation of cell proliferation, immune modulation,
angiogenesis, insulin clearance and organization of 3D tissue structure.
A retrospective study revealed a very strong association between the presence of
CEACAM1 on primary skin melanoma lesions and the development of metastatic disease
with poor prognosis. Over a time course of 10 years, almost 80% of the patients with
CEACAM1-positive primary tumor died, as compared to only 25% of the patients with
CEACAM1-negative primary tumors. The predictive value of CEACAM1 as a single
prognostic marker surpassed the most widely accepted marker, and is regarded as the
strongest predictive marker in melanoma. A similar prognostic association was reported in
non small-cell lung cancer, particularly lung adenocarcinoma. CEACAM1 is not expressed
by normal melanocytes.
We have demonstrated that CEACAM1 protects melanoma cells from elimination by NK
cells and by antigen-restricted T cells in vitro. Melanoma cells employ CEACAM1 to inhibit
effector functions such as killing and secretion of interferon-gamma. Blocking of
CEACAM1 did not induce non-specific killing of HLA-mismatched melanoma cells by
antigen-restricted T cells. Furthermore, we showed that melanoma cells are able to actively
elevate CEACAM1 expression, in response to interferon-gamma secreted from attacking
lymphocytes, to enhance their resistance to subsequent immune attack.
Our novel observations support the immune protective role of CEACAM1 in melanoma
cells and implies that the majority of metastatic melanoma patients might benefit from
CEACAM1-targeted therapy. We have generated a monoclonal antibody to CEACAM1 and
demonstrated in vitro and in vivo its potential therapeutic effect in various assays.
14
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Restoration of p53 anti-tumor activity by Zinc
IPK2 is a nuclear serine/threonine kinase that belong to a family of co-repressors of
homeodomain transcription factors and is considered as a central switch in the targeting of
cells toward apoptosis upon genotoxic stress by phosphorylating the oncosuppressor p53 at
Ser46. We have shown recently that hypoxia-driven HIPK2 downregulation can be
reversible. We found that zinc supplementation stabilizes and reactivates the hypoxiainhibited HIPK2, leading to repression of the HIF-1 pathway and restored p53-Ser46
phosphorylation and apoptotic activity.
Our study demonstrated that HIPK2-mediated transcription regulation includes inhibition of
HIF-1 whereas HIPK2 knockdown induced upregulation of HIF-1 pathway leading to
angiogenesis, chemoresistance and tumor growth in vivo. Additionally, it has been shown
that hypoxia-driven mechanisms inhibit HIPK2, creates a negative regulatory loop between
HIPK2 and HIF-1 that affects the multiple pathways regulated by both proteins, including
p53 and VEGF pathways.
Zinc supplementation rescues p53Ser46 phosphorylation and apoptotic activity as well as
HIPK2 activity. Our finding suggest a novel and unexpected way to reactivate hypoxiainhibited HIPK2 in cancer cells, and to exert its dual role in restraining tumor growth
through (i) inhibition of HIF-1 pathway, and (ii) through p53 activation in response to drug.
Furthermore, we provided evidence that this function may have an in vivo relevance by
improving chemotherapy response in treatment of tumor xenografts.
We have demonstrated that Intragastical Zinc chloride (ZnCl2) decreased tumor volume in
nude mice xenografts. Zinc restored p53 phosphorylation, conformation, and switches its
recruitment from oncogene to oncosuppressor genes in cell cultures.
The invention relates to methods for treating cancer by zinc compositions. Zinc
supplementation in vitro and in vivo reverses p53 phosphorylation and restores its tumor
suppressor activity.
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