Funding Research

MOCA has provided more than $7 million to ovarian cancer research projects since we were founded.

In 2017, we awarded $775,000 to ovarian cancer research projects in Minnesota.

Our goal: an early detection test, better treatments for women, and one day, a cure.

Promoting Ovarian Cancer Research

When it comes to ovarian cancer research funding, MOCA is a national leader among private, non-profit organizations.

Since 2001, MOCA has awarded more than $7 million in grants to professional researchers investigating various aspects of ovarian, fallopian and primary peritoneal cancer.

Every dollar directed to research means we’re getting closer to an early detection test, better treatments and, one day, a cure for ovarian cancer.

A panel of national expert reviewers help advise MOCA on the scientific merit and importance of each research proposal, along with a group of consumer advisory review advocates, made up of survivors and caregivers.

If you would like to learn more about MOCA’s research funding efforts, contact the MOCA office at (612) 822-0500.

A financial gift to MOCA helps ensure we can continue this vital work that impacts women today and future generations.

MOCA National Early Detection Research Grant Awards

We are excited to announce news of a milestone for MOCA’s ovarian cancer research funding efforts. Extending our commitment to funding the development of an early detection test for ovarian cancer, MOCA will be providing $100,000 in funding directed at a national research focused on early detection. In 2018, MOCA intends to award $50,000 to two projects focused on early detection from U.S.-based researchers from outside of Minnesota.

Application details, eligibility requirements and timeline can be found below.

Purpose

The Minnesota Ovarian Cancer Alliance will award $100,000 in November 2017 to support research on early detection of ovarian cancer.

Eligibility

Research proposals may be for individual projects or part of a larger research project related to early detection of ovarian, primary peritoneal or fallopian tube cancer. Awards will be granted only to projects conducted outside of the state of Minnesota. Project funding is available for 1 year of activity. Grant requests may be for a maximum of $50,000.

Selection

Evaluation of proposed projects will be made by national experts in early detection research, and at large MOCA members. Recommendations will be voted on by the MOCA Board of Directors.

Timeline

August 18, 2017:  Deadline for submission. All proposals must be received in the MOCA office by 4:00 pm Central Time on this date.

October 18, 2017:  Notification of action by MOCA.

Application Requirements

Proposals must be paginated and include the following:

  1. Cover letter should include the grant title, the PI’s name and direct contact information, total amount of funding requested,
  2. Abstract of proposed research, including rationale of the study (you do not need to include an explanation of why ovarian cancer is worthy of being studied or basic statistics on ovarian cancer as these are very familiar to all our reviewers), significance of research, explanation as to why this is innovative (if it is), aims, and a brief overview of research design and methods.
  3. Summary of proposed research (one paragraph) which describes the proposed research in terms an educated layperson would understand and describes the significance of the research to the patient population.
  4. Research proposal, not to exceed 6 pages. Additional supporting information may be provided in addendum form if deemed essential.
  5. Budget. Should clearly state total amount requested. Budget should include category of expense and include a rationale for each budget item. MOCA funds can be used for PI salary, conference travel and publication costs as deemed necessary by PI. Salary costs should be detailed. MOCA funds cannot be used for used for indirect costs.
  6. Information about PI, including:
    • Biographical statement of career goals of PI (one page)
    • Contact information, including direct phone number and email address
    • Curriculum vitae (resume), including education, honors, professional experience, publications, grants
  7. Letter of recommendation from the relevant person overseeing the institution, department, or lab where the research will be conducted.
  8. Project timeline detailing the expected chronological progress of research goals..
  9. Review of similar studies, including similar studies underway or pending which might influence subject accrual or funding.

Electronic proposals should be directed to kgavin@mnovarian.org and must be received by the MOCA office by 4 p.m. Central Time on August 18, 2017.

Send one original hard copy of the proposal to:

Kathleen Gavin
Executive Director
Minnesota Ovarian Cancer Alliance
4604 Chicago Avenue
Minneapolis, MN  55407

History of MOCA Research Grants

Since 2001, the Minnesota Ovarian Cancer Alliance has awarded more than $7 million in grants to professional researchers investigating various aspects of ovarian cancer at Minnesota institutions, including the University of Minnesota and Mayo Clinic.

In choosing which research projects to fund, special consideration is given to proposals that involve clinical trials and those that may lead to improvements in treatment for women with ovarian cancer.

Below is a listing of the Minnesota-based research projects which MOCA has funded in recent years. In all, MOCA has awarded more than 70 research grants.

2017 Ovarian Cancer Research Funding

In May 2017, MOCA awarded $775,000 to ovarian cancer researchers from the University of Minnesota and Mayo Clinic.

The 2017 funded researchers included:

  • Rachel Hurley, M.D., Ph.D. Candidate, Mayo Clinic. “Identifying novel therapeutic options for women with PARP inhibitor resistant ovarian cancer.” $50,000 for one year.
  • Aminah Jatoi, M.D., Mayo Clinic. “A phase 2 trial of auranofin and sirolimus for recurrent ovarian cancer.” $150,000 over three years.
  • Amy Skubitz, Ph.D., University of Minnesota. “Validation of biomarkers for the early detection of ovarian cancer.” $200,000 over two years.
  • Swayam Prabha, Ph.D., MBA, University of Minnesota. “Translational studies on glycoengineered mesenchymal stem cells in ovarian cancer.”  $100,000 for one year.
  • Andrea Wahner-Hendrickson, M.D., Mayo Clinic. ” PLK4 inhibition as a treatment strategy in high grade serous ovarian cancer” $275,000 over three years.

 

2016 Ovarian Cancer Research Funding

In May 2016, MOCA awarded a record $1,035,000.00 to ovarian cancer researchers at the University of Minnesota and Mayo Clinic.

The 2016 funded researchers included:

  • Daniel Vallera, Ph.D., University of Minnesota. “Engaging the Immune System to Attack Ovarian Cancer.” $96,908.00
  • Evanthia Galanis, M.D., Mayo Clinic. “Mesenchymal Stem Cell Delivery of Measles Virus for Recurrent Ovarian Cancer.” $89,636.00
  • Amy Skubitz, Ph.D., University of Minnesota. “New Platform for Multiplexing Biomarkers for the Early Detection of Ovarian Cancer.” $100,000.00
  • Marina Walther-Antonio, Ph.D., Mayo Clinic. “Ovarian Cancer Microbiome Signatures.” $98,335.00
  • Melissa Geller, M.D., M.S. University of Minnesota. “IL-15 Superagonist ALT-803 for Treatment of Advanced Staged Ovarian Cancer.” $100,000.00
  • Swayam Prabha, Ph.D., MBA. University of Minnesota. “Glycoengineered Mesenchymal Stem Cells for Targeting Platinum Resistant Ovarian Tumors.” $100,000.00
  • Martina Bazzaro, Ph.D., University of Minnesota. “Breaking Off Cancer Cells’ Addictions to Prevent and Treat Recurrent Ovarian Cancer.” $99,786.00
  • Kimberly Kalli, Ph.D., Mayo Clinic. “Identification of Ovarian Cancer Stromal and Stem Cell Antigens as Targets for Immunotherapy Designed to Prevent Ovarian Cancer Recurrence.” $70,000.
  • William Cliby, M.D., Mayo Clinic. “Investigating the Stromal Contribution to the Mesenchymal Subtype of Ovarian Cancer for Therapeutic Potential.” $70,000.
  • Cheryl Conover, Ph.D. and John Weroha, M.D., Ph.D., Mayo Clinic. “PAPP-A: A Novel Target for Chemo-resistant Ovarian Cancer.” $99,959.00
  • Reuben Harris, Ph.D. and Daniel Harki, Ph.D. University of Minnesota. “Stopping Enzyme-Catalyzed Ovarian Cancer Evolution.” $81,309.00

2015 Ovarian Cancer Research Funding

At MOCA’s Annual Meeting in May 2015, our organization provided a record $660,000 to research projects based at the University of Minnesota and Mayo Clinic. The projects are listed in the graph. This was the most funding ever awarded by MOCA at one time.

MOCA also branched out to national research funding for the first time in 2015, with $25,000 directed towards the Stand Up To Cancer (SU2C) Ovarian Cancer Dream Team funding, which MOCA has made a $100,000 commitment to be paid out over four years. 

The 2015 research projects include:

2015 WEB Grid

 

2014 Ovarian Cancer Research Grant Funding

2014_OV_Grants

2013 Ovarian Cancer Research Grant Funding

2013_OV_Grants

2012 Ovarian Cancer Research Grant Funding

2012_OV_Grants

2011 Ovarian Cancer Research Grant Funding

2011_OV_Grants

Ritonavir Repositioning from HIV-AIDS to Treatment of Ovarian Cancer

HIV-AIDS patients have very high prevalence of cancers. Protease Inhibitors (PI) are a class of drugs widely used to treat HIV-AIDS. In last two decades, numerous small and large observational studies have shown that patients using PIs have a drastic reduction in HIV-AIDS related cancers. These observations raised a question-do PIs have anticancer properties?

We took a widely used and well tolerated PI, Ritonavir (already FDA approved for HIV-AIDS and used worldwide) and tested it against several ovarian cancer cell lines and published that ritonavir exhibits potent anticancer activity against ovarian cancer in vitro. These anticancer effects are brought about by cell cycle arrest, activating apoptosis and inhibiting invasion and migration of ovarian cancer cells. Meanwhile other groups of investigators have reported similar data in other cancers (breast, lung, prostate and sarcoma) and have started clinical trials utilizing PIs (references included in grant application text).

Based on our robust published in-vitro data, we propose to conduct in-vivo studies to treat ovarian cancer in mice with ritonavir in the present MOCA grant application. If funded, completion of the in-vivo mice studies will pave the way to start human trials of ritonavir in ovarian cancer.

Lymphodepleting Chemotherapy and T-Cell Suppression Followed by Allogeneic Natural Killer Cells and Interleukin-2 in Patients with Recurrent Ovarian Cancer

Our preliminary clinical Phase II study at the University of Minnesota using allogeneic (derived from healthy donors) NK cell infusions to treat ovarian cancer has shown exciting potential clinical efficacy with the ability to detect donor-derived NK cells up to 35 days following infusion. It is from this observation that stems our current clinical trial and ongoing basic science research which addresses how to optimize NK cell expansion, which we hypothesize, is required for therapeutic efficacy. To understand the mechanism of NK mediated tumor cell killing and NK cell expansion we will create Green Fluorescent Protein (GFP) expressing MA148 and OVCAR3 mouse models to test clinically relevant endpoints.

Two aims were chosen, each of which will provide better insight into how to proceed with our clinical trial

Aim 1.To determine whether intravenous (IV) or intraperitoneal (IP) delivery of activated NK cells is the optimal route of delivery to promote ovarian cancer cell kill.

Aim 2. To determine if extended dosing of bortezomib in combination with IL-2 activated allogeneic NK cells can potentiate cell kill in a mouse model of ovarian cancer by:

a.) sensitizing ovarian cancer to Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL) and Fas mediated NK cell killing
b.) decreasing tumor MHC-1 expression
c.) decreasing tumor volume

Sub Aim 2a. To determine the above in a mouse model of platinum sensitive ovarian cancer
Sub Aim 2b. To determine the above in a mouse model of platinum resistant ovarian cancer

Specifically, in both aims we will be testing tumor cell kill and tumor infiltration by NK cells with and without bortezomib treatment. The outcomes of this research will have significant implication because we expect the knowledge will suggest development of novel immunotherapy treatment strategies and therapeutic interventions needed for women with ovarian cancer. The research will be of additional significance, because what is learned will contribute to the broader understanding of how immune based therapies can be utilized as an approach to treatment in women with this devastating disease. My preliminary clinical data and in vitro and mouse model findings demonstrate the importance of integrating these entities to understand novel clinical strategies and to improve efficacy.

Targeting the Tumor Microenvironment in Ovarian Cancer

Following optimal cytoreductive surgery, ovarian cancer patients are commonly treated with carboplatin/cisplatin and paclitaxel [3], with only ~10 15% of >stage 3 patients remaining disease-free at five years. The rest ultimately develop chemotherapy-resistant disease, relapse, and eventually succumb – often within 1-2 years of relapse. Hence, there is considerable need to develop improved up-front and salvage treatment strategies for ovarian cancer. Although various targeted therapeutics have been explored that include monoclonal antibodies directed towards vascular endothelial growth factor , VEGF (bevacizumab) and folate receptor, small molecule tyrosine kinase inhibitors, PARP and mTOR inhibitors, none of them specifically target the tumor microenvironment and inhibit several pathways mediated by various growth factors simultaneously. Extracellular matrix (ECM) heparan sulfate (HS) glycosaminoglycans are complex polysaccharides that are ubiquitous in nature and play important roles in the regulation of several aspects of cancer biology, including angiogenesis, tumor progression and metastasis. We are proposing to test the efficacy of a novel compound, a HS mimetic, PG545 that has never been tested in ovarian cancer that targets heparanase and heparin binding growth factors overexpressed in the tumor microenvironment. PG545 has been shown to inhibit solid tumor progression of breast, prostate and liver cancer xenografts. This selective inhibition of the heparanase enzyme influences cellular extravasation through the basement membranes and remodels tissues during tumor growth. More importantly, heparan sulfate proteoglycans, the substrate for heparanase act as coreceptors for pro-angiogenic and pro-metastatic growth factors such as FGF2 and VEGF.

Therefore targeting HSPG- mediated signaling using heparan sulfate (HS) mimetic is a very innovative approach.

Hypothesis and Objectives: We hypothesize that HS mimetics that target heparanase and multiple growth factors in the tumor microenvironment will function as an inhibitor of multiple signaling pathways simultaneously resulting in inhibition of tumor growth, angiogenesis and metastatic spread. Based on our in vitro preliminary data presented in figure 2 that indicate increased cisplatin -induced cytotoxicity when combined with PG545, while PG545 alone had minimal anti-proliferative effects indicating synergy, we further hypothesize thatPG545 might represent a candidate ovarian cancer therapeutic either as a single agent or in combination with cytotoxic chemotherapy. The objective of the proposed study is to test the efficacy of the HS mimetic PG545 in inhibiting angiogenesis and peritoneal metastasis in vivo both alone and in combination with cisplatin and/or paclitaxel using two different ovarian cancer models.

Specific Aims and Study Design: In order to test our hypotheses, we propose three specific aims.

In Aim 1, we will assess the in vitro effects of PG545 in potentiating cisplatin and or paclitaxel induced cytotoxicity in several ovarian cell lines using colony formation assays (CFA), and cell cycle analysis by FACS.

In Aim 2, we will examine of the in vivo efficacy of PG545 in potentiating cisplatin and or paclitaxel induced cytotoxicity of ovarian xenografts in athymic immunodeficient mice and examine the effect of PG545 on angiogenesis by Matrigel Plug Assay. While experiments in immune-compromised mice bearing human cancers have merit, such models ignore effects of PG545 that may require either i) an intact immune systems or ii) syngeneic tumor-stroma interactions.

Therefore in Aim 3, we will assess the therapeutic efficacy of PG545 in a syngeneic ID8 murine ovarian cancer model.

Innovation: Due to the highly refractory nature of recurrent ovarian tumors, standard treatment options lead to short term survival rates. Thus, the pursuit of novel therapeutics is of utmost importance if treatment advancements are to be made to improve overall survival and disease free survival of patients with ovarian cancer. With this award, we will establish whether one of the novel agents, HS mimetics, that is known to inhibit heparanase and growth factor mediated signaling are also involved in modulating chemoresponse and or metastatic spread of cancer. Despite advances of testing multiple targeting agents in ovarian cancer, there are limitations with each one of the targeting agent due to the lack of understanding of redundant pathways that exist in cell signaling and the mechanism of resistance. There are no preclinical studies of PG545 in ovarian cancer. Testing the effect of PG545 in cell lines both in vitro and in vivo is innovative and will provide important preclinical data on the potential use of PG545 in future clinical trials.

Impact: Our preliminary data suggests that PG545 targets multiple pathways simultaneously and thus may have a better advantage over other targeting agents since the target also includes heparanase, an extracellular enzyme that is over expressed in ovarian cancer and is associated with poor prognosis. Based on the report that HS mimetics sensitize cells to chemotherapy induced cytotoxicity in patients with prostate cancer lends additional support to testing this agent in preclinical models of ovarian cancer. If HS mimetics are successful in inhibiting carcinomatosis and sensitize cells to drug induced cytotoxicity, the impact of this research to patients with ovarian cancer will be substantial.

Novel Bispecific Ligand-Directed Toxins for the Treatment of Ovarian Cancer

Ovarian cancer is the most lethal malignancy of the female reproductive tract. While most women will achieve complete remission after treatment (surgery and chemotherapy), the majority will relapse within two years, highlighting the need for novel therapies. Cancer stem cells (CSC) have been identified in a number of different types of solid tumors (including ovarian cancer) as a small population of cells that are relatively quiescent, can self-renew, form “spheroids” or multicellular aggregates when grown on a nonadhesive surface, and maintain the tumor by differentiating into cells that make up the tumor bulk. Since conventional chemotherapies target only the rapidly dividing “bulk tumor” cells, CSC may be the primary source of tumor recurrence.

The hypothesis driving this study is that ovarian cancer tumors can be eradicated by use of liganddirected toxins that specifically target each of the two populations of cancer cells within the tumor simultaneously; in effect eliminating both the CSC and the bulk tumor cells and thereby preventing recurrence of the disease. To this end, several ovarian cancer cell lines will be evaluated for the presence of CD133, a CSC marker, on their cell surface. A novel specific CD133-directed toxin will be tested for its efficacy in killing ovarian cancer cells in vitro and in vivo, alone and in combination with other monospecific and bispecific ligand-directed toxins that target the more differentiated tumor cells.

Aim #1 is to characterize the expression levels of CD133 on the surface of several ovarian cancer cell lines and normal surface epithelial cell lines by flow cytometry. CD133+ and CD133- cells will be segregated and collected by fluorescent activated cell sorting.

Aim #2 is to correlate the expression of CD133 with the CSC phenotype by perform in vitro functional assays on the CD133+ and CD133- cell populations. In addition, CD133+ and CD133- cell populations will be tested for their tumorigenicity in nude mice.

Aim #3 is to test the sensitivity of CD133+ and CD133- cells to our novel CD133-directed toxin (dCD133KDEL), as well as other monospecific and bispecific ligand-directed toxins vs. EGFR, IL-4R, IL-13R, and uPAR that we developed and found to be successful at eradicating other solid tumors.

Aim #4 is to test the CD133+ and CD133- cells for their sensitivity to the toxins from Aim #3 in an in vivo mouse model.

The results of this study will shift the paradigm for ovarian cancer treatment from one that treats patients solely with chemotherapy that targets the rapidly proliferating cells to one that targets both the rapidly proliferating “bulk tumor” cells and the quiescent CSC. This will have a significant impact on disease recurrence and thereby increase the rate of survival for ovarian cancer patients.

2010 Ovarian Cancer Research Grant Funding

2010_OV_Grants

Generation of Clinically Relevant Ovarian Cancer Xenograft Models

Ovarian cancer is the most lethal malignancy afflicting women in the U.S.  One of the main barriers to improving the survival of patients with ovarian cancer is indentifying novel agents that will improve existing therapy.  This barrier is the result of the heterogeneity of epithelial ovarian cancers (EOC) and the lack of models that accurately predict the sensitivity of ovarian cancers so systemic therapies.  Currently, the researchers are developing a living tumor bank of intraperitoneal models of EOC, allowing them to generate tumor models in severely immunocompromised (SCID) mice from individual EOC patients having their primary debulking surgery performed at the Mayo Clinic.  The researchers hope to expand the models to generate sufficient tumor mass from each founder line.  This expansion will establish sufficient materials to compare the expression profiles of established models to the source patient and re-establish tumor materials for mice for testing novel and existing therapies in vivo.  The researchers will attempt to validate their tumor models by comparing the benefits of chemotherapy with and without insulin-like growth factor (IGF) pathway targeting in SCID mouse models and the source patients who are receiving IGF-targeted therapy as part of their front-line phase II clinical trial, TRIO 014.  The clinical efficacy of treatment on TRIO 014 will ultimately be compared to the efficacy of IGF targeting in models from the source patient.  Specifically their aims are: (1) expand in vivo founder models of ovarian cancer/primary peritoneal carcinoma derived from patient tissue; (2) compare the expression profile of tumor from expanded models to source patient tumor; and (3) explore the efficacy of insulin-like growth factor targeting between matched patient and in vivo models.

Maintenance Treatment with Metformin in an In Vivo Ovarian Cancer Model

In their study, the researchers purpose to investigate metformin as a drug to limit recurrence of ovarian cancer and prolong survival.  Metformin is a well tolerated FDA-approved drug and is currently the single most prescribed oral anti-diabetic agent for type2 diabetes.  Preliminary data done by the researchers suggests that: (1) metformin inhibits ovarian cancer cell proliferation by activating LKB1-AMPK pathway; (2) induces cell cycle arrest accompanied by decreased cyclin D1 and increased p21 expression; (3) inhibits mTOR-S6SP –protein translation and (4) lipid biosynthesis; (5) significantly inhibited the growth of A2780-induced ovarian cancer tumors in nude mice; (6) significantly enhance the cytoxic effect of cisplatin to attenuate clonogenic survival in A2780 and its resistant derivative cell line, C200; and (7) significantly enhanced cytotoxicity of cisplatin in vivo.  The specific aim of this research study is as follows: to determine the preclinical efficacy of metformin in maintenance therapy to prevent or delay the recurrence and prolong survival of ovarian cancer in vivo.

Activated Natural Killer Cells and Proteasome Inhibition in Ovarian Cancer

Exciting new research is beginning to elucidate the connections and interactions between the elements of the innate immune system and to describe their role in tumor surveillance and killing.  The ability to manipulate and direct the innate immune system to improve the specificity and intensity of its anti-tumor activity is an urgent goal of immunotherapy research.  Preliminary clinical Phase II studies done by the researchers at the University of Minnesota using allogeneic (derived from healthy donors) natural killer (NK) cell infusions to treat ovarian cancer has shown exciting potential clinical efficacy.  Prior research shows that NK cells derived from ovarian cancer patients are defective in killing tumor.  The researchers’ data shows that IL-2 activated allogeneic NK cells can kill ovarian cancer cells in vitro and that in a mouse model of ovarian cancer the addition of both the Proteasome inhibitor bortezomib and extended IL-2 are needed for decreased tumor growth and upregulation of the biomarker fas.  It is from these observations that stem their current clinical trial and ongoing basic science research.

There are two specific aims of this research study:

  1. to determine if extended dosing of bortezomib in combination with IL-2 can increase infiltration of IL-2 expanded allogeneic NK cells into ovarian cancer tumors; and
  2. to determine if extending dosing of bortezomib in combination with IL-2 expanded allogeneic NK cells can (a) sensitize ovarian cancer to Tumor Necrosis Factor-Related Apoptosis-inducing Ligand (TRAIL) and Fas mediated NK cell killing and (b) decrease tumor MHC-1 expression.

2009 Ovarian Cancer Research Grant Funding

2009_OV_Grants

Characterizing the Effects of Sulfatase Inhibition on Ovarian Cancer in Vitro and in Vivo

Recently, the researchers demonstrated that the steroid sulfatase, the enzyme necessary to convert ES to low potency estrone (E1) and some forms of 17 beta hydroxysteroid dehydrogenase (17β-HSD) which convert E1 to biologically potent estradiol E2, were present in virtually all of 100 ovarian cancer specimens surveyed.  The researchers were able to further demonstrate that patients with high sulfatase activity tumors, that is those most capable of generating an active growth factor, had significantly worse progression-free and overall survival despite controlling for al other known risk factors (age, stage, grade, histology, surgical outcome, and platinum sensitivity).

In their current study, the researchers aim to evaluate in vitro and in vivo the effects of inhibiting the sulfatase and aromatase pathways in ovarian cancer cells.  Ultimately the goal of these experiments is to gain the insight necessary to optimally apply evolving, low-toxicity endocrine therapies to women with ovarian cancer.  It is possible that insight gained herein may ultimately be used to develop a more directed prevention strategy than estrogen-based oral contraceptives, which are the dominate paradigm today.  The researchers have two specific aims for their study; (1) to characterize the effect of modulating the sulfatase pathway on ovarian cancer cells in vitro; and (2) to characterize the molecular and clinical effects of sulfatase inhibition on established ovarian cancer tumors in vivo using a nude mouse model.

Blockade of PD-1 during Vaccination against Ovarian Cancer

Ovarian cancer is associated with high mortality rate in women and it was estimated that 15,210 women died of ovarian cancer in 2008.  Even though there were strategies such as surgery and chemotherapy using platinum based paclitaxel for the treatment of ovarian cancer, their success is minimal, with the majority of ovarian cancer patients being at risk of recurrence of disease.  It is well known that cancer patients have pre-existing immunity against different tumor associated antigen.  To supplement surgical and chemotherapeutic strategies, immunotherapy using tumor associated antigen based vaccines to boost anti-tumor immunity was also attempted.  Limited success was achieved and the role of complex immune suppressive network of ovarian cancer was attributed to this failure.  Thus, it would be ideal to design a strategy targeting the immune suppressive network and using TAA based peptide vaccine simultaneously for the treatment of ovarian cancer.

We identified that PD-1+ DC, a novel component in tumor microenvironment, is inhibitory to tumor specific immune responses and can be targeted using anti-PD-1 monoclonal antibody.  We also indentified that ovarian cancer patients have endogenous immunity against a tumor associated antigen called folate receptor alpha (FRα) and used the immunogenic epitopes of FRα as peptide vaccine in pre-clinical models.  In this proposal, we would like to determine the characteristics of PD-1+ DCs and improve the therapeutic efficacy of FRα peptide vaccine by targeting the suppressor of PD-1+ DCs in a preclinical mouse model (ID8 model).  The researcher’s long term goal is to test current approaches in human clinical trails after achieving encouraging results in the proposed study.  The specific aims  of this research are (1) To determine to role of PD-1+ DC in the ovarian cancer immune microenvironment; and (2) to determine the effect of combination therapy using anit-PD-1 antibody and folate receptor alpha (FRα) peptides as a vaccine in an immunocompetent mouse model of human ovarian cancer.

Defining the Nuclear Progesterone Receptor as an Anti-Proliferative, Pro-Apoptotic Tumor Suppressor of Ovarian Cancer

Despite advancements in screening, detection, surgery, and chemotherapy, ovarian cancer remains the most lethal form of gynecological malignancy.  While its etiology is unknown, at least 20-40% of ovarian cancers express steroid hormone receptors and hormone-mediated signaling has been shown to regulate ovarian cancer cell proliferation and survival.  In particular, published reports repeatedly suggest that the ovarian steroid hormone, progesterone, may be an inhibitor of proliferation and/or pro-apoptotic to ovarian surface epithelial cells. Additionally, our group and others have shown low to undetectable levels of the nuclear progesterone receptor (PR) in cell line models of advanced ovarian cancer, suggesting that progesterone’s tumor-suppressive properties are selected against during the process of malignant transformation.  Progesterone is clearly mitogenic in breast cancer models.  However, forced expression of the PR induced p21 expression and ovarian cancer cell senescence in the presence of progestin; cells were growth-inhibited but very long-lived.  Notably, these long-lived senescent cells were sensitized to undergo apoptosis induced by chemotherapeutic protein kinase inhibitors.

Thus, the researcher hypothesized that the nuclear PR is a novel ovarian cancer tumor suppressor.  Re-expression of the PR in ovarian cancer cells confers anti-proliferative effects and greatly increased sensitively to pro-apoptotic agents.  To test this hypothesis, the researchers propose two aims to (1) define how PR activity mediates the senescent phenotype in ovarian cancer cells, and (2) uncover reversible molecular signaling mechanisms that lead to the repression/re-expression of the PR in advanced ovarian cancer cells.  As a result, the researchers hope to gain a more thorough understanding of the molecular mechanisms underlying ovarian cancer progression.

Pilot Study of Hyperthermic Intraperitoneal Chemotherapy Utilizing Carboplatin in First Recurrence Ovarian Cancer

Standard intraperitoneal chemotherapy (IP) has revolutionized the treatment of advanced stage ovarian cancer in recent years.  This is likely due to the increased tumor to plasma chemotherapy ratio and exposure time.  The addition of hyperthermia (heat) to the process of IP has been shown to further enhance the permeability of chemotherapy within tumor cells as well as reverse some acquired resistance in the recurrent setting.  The researchers therefore seek to apply this theoretical benefit of delivering hyperthermic intraperitoneal chemotherapy (HIPC) to patients with recurrent ovarian cancer.  Through this study, the researchers seek to determine the feasibility and safety of delivering HIPC in this setting.  They will further determine the impact of HIPC on progression free survival, clinical response, and overall survival in women with recurrent ovarian cancer.  The researchers propose three specific aims in their research; (1) to determine the feasibility and safety of delivering hyperthermic intraperitoneal chemotherapy (HIPC) in the recurrent setting; (2) determine the clinical response, disease free survival, and overall survival in patients treated with adjuvant HIPC at the time of first clinical recurrence of ovarian, fallopian tube, or primary peritoneal carcinoma in platin sensitive patients; and (3) to monitor for toxicities, complications and changes in quality of life measures associated with HIPC procedures.

2008 Ovarian Cancer Research Grant Funding

2008_OV_Grants

Targeting a Novel Membrane Progesterone Receptor (mb-PR) Overexpressed in Human Ovarian Cancer

Ovarian cancer remains the most lethal form of gynecological cancer.  Published reports suggest that the ovarian cancer steroid  hormone, progesterone (P4), may be growth inhibitory and proapoptotic in ovarian surface epithelium, and thereby protect women from ovarian cancer development.  The actions of P4 are thought to e mediated by the nuclear progesterone receptor (n-PR) A and B isoforms, which act as ligand-activated transcription factors capable of regulating gene expression by direct or indirect binding to DNA promoter elements.

Recently, however, a novel cell membrane-localized progesterone receptor (mb-PR) was cloned from spotted sea trout ovaries, and mb-PRs encode distinct G-protein coupled receptors capable of regulating multiple intracellular signaling pathways in response to P4, including mitogen-activated protein kinases (MAPKs) and cyclic AMP (cAMP)-dependent protein kinases (PKA).  This study aims to (1) Define the expression and regulation of n-PR relative to novel mb-PR in ovarian cancer cell lines and tissues; (2) Define the signaling pathways activated in response to mb-PR; and (3) Define the mechanism(s) and biology of mb-PR apoptotic signaling.  AS a result, we hope that newly defined mb-PR-dependent signaling pathways may be exploited in ovarian cancer treatment as a means to sensitize ovarian cancer cells to cell death-including chemotherapeutics, perhaps as part of combination therapies aimed at reducing toxicities and improving clinical outcome.

Regulation of Hsulf-1 Expression by vHNF(TCF2): A Marker for Ovarian Clear Cell Carcinoma

Of the approximately 27,000 new cases of ovarian cancer diagnosed annually in the U.S., 14,000 will ultimately prove fatal. An improved understanding of the alterations associated with the development and progression of this cancer could conceivably allow the development of early detection markers and alternative therapeutic approaches.  A better understanding of genetic alterations underlying the development of ovarian cancer may provide vNHFs for critical understandings of the disease process.  To search for these alternations, we generated suppression subtraction hybridization (SSH) cDNA libraries from 4 primary tumors.  One of the differentially expressed genes identified from this screen was HSulf-1, a human homolog of QSulf-1.

This study plans to define the role of HSulf-1 downregulation in the behavior of ovarian cancer and initiate a search for agents that might target vHNF to reverse the effects of HSulf-1 downregulation.  If successful, these experiments will characterize a novel mechanism of drug resistance in cancer.  To further evaluate the biological effects of HSulf-1 loss regulated by vHNF, the research proposes to (1) Validate HSulf-1 as a vHNF target gene; (2) Determine if downregulation of vHNF contributes to cisplatin induced cytotoxicity; and (3) Determine whether HSulf-1 expression correlates with loss of vNHF in patient tumors and correlate with clinical behavior of ovarian cancer.

Ovarian Cancer Biomarker Discovery by Proteomic Techniques

This study is focused on improving the detection of ovarian cancer by identifying novel biomarkers for ovarian cancer.  To do this, the researchers will analyze the protein profiles of two new sources of ovarian cancer biomarkers which they believe to be more enriched for ovarian cancer-specific proteins.  Namely, they propose to analyze the spent media of ovarian cancer cell lines and the ascites fluid of ovarian cancer patients.  There are four aims of this research (1) To grow ovarian cancer cell lines and normal ovarian surface epithelial (NOSE) cell lines in serum-free media and collect the spent media, which will contain the proteins secreted by the cells; (2) Isolate the “low abundance” proteins from ovarian cancer ascites samples and “non-ovarian cancer” ascites samples by affinity chromatography on a ProteomeLab ™ IgY12 HPLC colum, follwed by a GenWay SuperMix HPLC column; (3) Identify new biomarkers for ovarian cancer by Differential In Gel Electrophoresis (DIGE), in which two-dimensional gel electrophoresis will be used to simultaneously analyze proteins from ovarian cancer samples and “non-ovarian cancer” samples in order to indentify proteins that are different between the sample sets.  Proteins will then be indentified by mass spectrometry; and (4) Confirm and/or expand upon the differentially expressed biomarkers by iTRAQ™ methodology in which liquid chromatography coupled with mass spectrometry will be used to simultaneously analyze proteins.

Genomic, Genetic, and Functional Studies in Ovarian Cancer

Epithelial ovarian cancer (EOC) is the most lethal gynecological malignancy in the United States.  Major challenges in ovarian cancer are the need for improved understandings of early and late genetic alternations so that biomarkers and novel therapeutic targets can be discovered in ovarian cancer.  Identification of these genetic alterations is essential in our effort to discover genetic markers for early-stage ovarian cancer and to overcome the challenge of chemoresistant ovarian cancer.

The objective of this study is to identify altered transcripts and pathways that are associated with early and late-stage serous ovarian cancer, and to indentify novel therapeutic targets in chemoresistant tumors.  To accomplish this, the study aims to (1) Characterize transcripts in serous ovarian tumors by high-throughput sequencing; (2) Indentify transcripts associated with early-stage ovarian tumors; and (3) Identify transcripts associated with chemoresistant phenotype.

Targeting the Mullerian Inhibiting Substance Receptor for Therapy and Imaging in Ovarian Cancer

Most patients with epithelial ovarian cancer (EOC) will not be cured by conventional treatment.  Primary obstacles to cure are advanced stage, emergence of chemo-resistance and dose-limiting toxicity from normal tissues. Specific targets for EOC are desperately needed.  In the present application we evaluate the feasibility of using the Mullerian inhibiting substance (MIS) type II receptor as one such target.  MIS is a member of the TGF-β superfamily whose biological actions require both type 1 (T1R) and type II (T2R) receptor. While the T1R for MIS is controversial, the MIS T2R (MISRII) is expressed on the majority of EOC as we have published utilizing funding from a prior grant from MOCA.  The general hypothesis to test is that MISRII is a tissue-specific receptor on the surface of most EOC which, additionally, is fundamentally linked to a signaling pathway(s) resulting in growth inhibition and/or cell death.  The aims of this study are (1) Assess the ability to target MISRII expressing ovarian cancer cells in vitro; and (2) Demonstrate effective and specific targeting of MISRII expressing human EOC xenografts in the mouse model of EOC.

2007 Ovarian Cancer Research Grant Funding

2007_OV_Grants

Validation of Potential Biomarkers for Ovarian Cancer discovered by DIGE and iTRAQ ™

The studies outlined in this proposal are focused on improving the detection of ovarian cancer by validating novel biomarkers for ovarian cancer.  Early detection of ovarian cancer results in improved survival rates for patients.  Since current screening techniques are neither adequately sensitive nor specific, ovarian cancer is usually not detected in its early stages.  In addition, novel biomarkers are needed for detection of recurrence of ovarian cancer.  Advances in technology and state-of-the-art instrumentation have allowed us to combine the highly sensitive instrumentation of mass spectrometry with conventional gel electrophoresis and liquid chromatography.  During the past year, we have successfully used to complementary proteomics techniques to discover five novel biomarkers for ovarian cancer.

Our long term goal is to develop a more highly sensitive and specific diagnostic assay for ovarian cancer.  During the course of this one-year study, we will conduct the following experiments: (1) We will validate the presence of the five protein biomarkers in the sera of ovarian cancer patients; (2) We will quantitative the levels of expression of the five protein biomarkers in the sera of ovarian cancer patients; and (3) We will determine whether the five protein biomarkers are expressed and secreted by ovarian cancer cell lines, ovarian cancer tissues, and/or ovarian cancer cells from ascites.

Candidate Gene Approach for Identification of Novel Ovarian Cancer Susceptibility Genes

Despite profound increase in our understanding of the genetic basis of ovarian cancer in the decade since mapping of BRCA1, it has become evident that additional genes remain to be found.  Although BRCA1 and BRCA2 account for the majority of site-specific ovarian and breast-ovarian cancer families, approximately 45-70% of ovarian cancer cases within significant family history are not attributable to these genes.  Given that at most 20% of familial risk is attributable to BRCA1 and BRCA2 and that the majority of women referred to family cancer clinics do not carry a mutation in either gene, identification of the remaining ovarian cancer susceptibility genes is clearly a problem of primary importance in ovarian cancer research.  The researchers purpose to apply direct analysis of biologically plausible candidates for genes for mutations in high-risk non-BRCA1/BRCA2 ovarian cancer families in order to overcome the intrinsic limitations of conventional gene identification efforts.

Through the use of empirical observation, it will be possible to indentify genes in which mutations contribute to only a minor fraction of hereditary ovarian cancer.  There are two specific aims of the research; (1) conduct a comprehensive mutation screen of familial ovarian cancer cases for mutations in the BRCA1, BRCA2 or the MSH2, MLH1 and MSH6 mismatch repair (MMR) genes to verify that genetic predisposition of these patients I not attributable  to any of the known susceptibility  genes (non-B1/B2/MMR).  Familial ovarian cancer cases are defined as having a family history in first and second degree relatives (proband inclusive) of 2 or more ovarian cancer cases; and (2) mutation screening of non-B1/B2/MMR familial ovarian cancer patients of 30 candidate genes, selected on the basis of biological evidence supporting a role in the same biological pathways as BRCA1, BRCA2 or the mismatch repair genes.

Pre-clinical Evaluation of a Folate Peptide Vaccine for Ovarian Cancer

The long-term goal of this research program is to determine whether a combination approach of overcoming tumor-induced immunosuppression and boosting immunity against the small volume disease that remains in some women after surgery and chemotherapy can delay or prevent disease recurrence.  Initially, we purpose to deplete cells that suppress immune responses prior to immunizing women with a peptide vaccine targeting FRα.  In order to obtain approval form the Food and Drug Administration (FDA) to perform this Phase I clinical trial, we must provide evidence that the peptides chosen for the vaccine are found as naturally processed peptides in humans and show safety and efficacy data in an animal model.  Accordingly, our specific aims are: 1) to determine if human FRα-derived HLA class II peptides are naturally processed; and 2) to determine if murine FRα peptide vaccines are therapeutically effective against mouse FRα-expressing ovarian tumors.  With the data obtained in this proposal, we will submit the Investigational New Drug application describing the human multi-peptide vaccine to the FDA.  The Phase I trail to determine whether the vaccine is safe and effective at inducing immune responses against FRα is the critical first step towards immunotherapeutic approaches to delaying or preventing ovarian cancer recurrence.

Role of Serine Protease HtrA1 in Targeting sCLU implicated in Chemoresistance

Most women with advanced ovarian cancer ultimately relapse and die of disease progression despite an initial response to first line platinum-paclitaxel; and only a small minority (~20%) is cured with the standard combination.  Identification of genes that confer chemoresistance and/or chemosensitivity will aid in the identification of patients responsive to specific chemotherapy regimens.  A deeper understanding of the underlying biology of these tumors, and the development of new “biological” therapeutics that allow a more efficient and durable response may lead to tailored therapies in patients with ovarian cancer.

This study aims to (1) Determine if Clusterin is a substrate of HtrA1 in ovarian cancer; (2) To evaluate the effect of sCLU expression on ovarian cancer chemoresistance in vitro; and (3) Correlate the expression of sCLU to chmeoresponse, disease free survival and overall survival in patient samples.

Neoadjuvant Chemotherapy in Ovarian Cancer: Predicting Tumor Response

Epithelial ovarian cancer has the highest mortality rate of women’s cancers with approximately 16,210 deaths estimated in the U.S. last year.  Despite aggressive surgical approaches and combination chemotherapy, most women with advanced disease ultimately relapse and die of their disease.  Unlike other cancers, 75% of women presenting with advanced ovarian cancer respond to paclitaxel and carboplatin but only 20-30% will be alive at 5 years.  Surgical cytoreduction followed by paclitaxel and carboplatin chemotherapy is the standard of care for treatment in this disease.  Patients who present with extensive tumor burden not amenable to surgical cytoreduction or those who are too ill to undergo a radical cytoreductive surgery have only neoadjuvant chemotherapy or palliative care as options.  Neoadjuvant chemotherapy allows for a short course of initial chemotherapy with an opportunity to take the majority of these women to the operating room to assess response to chemotherapy and disease remaining.

The population of women addressed in this study will give us the opportunity to analyze tumors before and after their first line of chemotherapy, a population in which the tumor burden is so extensive or because of existing co-morbidities, surgery is not an option.  Examining gene expression profiles, biomarkers and tumor resistance patterns before and following chemotherapy in this population of women will lead to identification of prognostic indicators to determine who is at highest risk for not responding to neoadjuvant chemotherapy and to explore changes in biomarkers and gene expression that have occurred during that short time of chemotherapy exposure allowing the tumor to persist.  The objective of this application is to clinically explore the relationship between tumor biomarkers, gene profiles and drug resistance testing in women with advanced ovarian cancer who are not able to undergo an initial surgical cytoreductive surgery.

The central scientific hypothesis is that exposure to chemotherapy selects out chemotherapy resistant cells that overexpress certain biomarkers and contain newly acquired DNA mutations which can translate into altered gene expression and mutated proteins.  Three trial aims were chosen, each of which will provide unique training opportunities in translational research in support of my research objectives.  1. To determine if biomarkers predictive of poor outcome will be up-regulated following exposure to chemotherapy in patients who respond poorly to chemotherapy.  2. To determine if chemotherapy selects for tumor cells with newly acquired DNA mutations which translate into altered gene expression, therefore changing tumor gene expression profiles following treatment with chemotherapy.  3. To determine if the presence of biomarkers, previously associated with poor outcome, and extreme drug resistance (EDR) assays can predict the ability to perform optimal cytoreduction in this patient population.

2006 Ovarian Cancer Research Grant Funding

2006_OV_Grants

Complementary Proteomic Techniques to Identify Ovarian Cancer Biomarkers

The studies outlined in this proposal are focused on improving the detection of ovarian cancer by identifying novel biomarkers for ovarian cancer. Early detection of ovarian cancer results in improved survival rates for patients. Since current screening techniques are neither adequately sensitive nor specific, ovarian cancer is usually not detected in its early stages. In addition, novel biomarkers are needed for detection of recurrence of ovarian cancer.

The field of proteomics encompasses the next generation of discovery tools for those seeking biomarkers of disease. Advances in technology and state-of-the-art instrumentation have combined the highly sensitive instrumentation of mass spectrometry with conventional gel electrophoresis and liquid chromatography.

A major roadblock in the discovery of protein biomarkers that are disease-specific are the abundant proteins of serum. Just 12 proteins make up approximately 90% of the serum. Thus, unique proteins that may be specifically found in the sera of women with ovarian cancer are masked by these highly abundant proteins.

To overcome this problem, we propose first to remove these 12 highly abundant proteins from all sera samples, and then to analyze the samples by three different techniques in order to identify ovarian cancer-specific proteins. The three mass spectrometry-based techniques that we propose to use are highly complementary, such that the deficits of one technique are overcome by the strengths of the others. Furthermore, the data generated by the three techniques will permit better adjustment of the sample preparation and analysis protocols to optimize the ability to discover specific protein biomarkers in the sera of patients with ovarian cancer.

Defining the Mechanism of Drug Resistance in Ovarian Cancer

Most women with advanced ovarian cancer ultimately relapse and die of disease progression despite an initial response to first line platinum-paclitaxel; and only a small minority (~20%) is cured with the standard combination. If those women who will drive little benefit from standard platinum-paclitaxel could be identified prior to therapy, namely those with early recurrence, then early intervention with alternative approaches, including novel agents targeted to molecular pathways (that are aberrantly activated), may lead to tailored therapies in patients with ovarian cancer.

In this context, a recent report from Tweari et al that analyzed 6,990 malignant tissues from primary and metastatic sites using the extreme drug resistance assay suggested that the molecular alterations that lead to drug resistance may occur early in the carcinogenic process and persist in the metastatic sites. Therefore, the identification of genetic changes in the tumor at the time of diagnosis after the debulking surgery may help clinicians in choosing a targeted therapy.

Toward this goal, we utilized cDNA microassay technology containing 30,721 transcripts to profile gene expression patterns from 79 ovarian cancer specimens to identify a predictive model for those tumors with early recurrence. These analyses resulted in the identification of 14-gene model that accurately predicted early or late recurrence in 86% of patients in the test set (p<0.05). Interestingly, the list did not include genes known to be involved in drug resistance such as ATP binding cassette transporters, metallothionine or DNA repair components. Instead, our analysis identified a unique set of nuclear factors that are implicated in survival and programmed cell death.

The anti-proliferative effects of cisplatin and other genotoxic chemotherapeutics are thought to be exerted by engagement of apoptotic regulators. The dysregulation of these pathways in cancer cells not only confers and intrinsic survival advantage, but also renders cells chemoresistant. In ovarian cancer, drug tailored regimens are hindered by a lack of understanding as to exactly how drug treatments engage apoptotic and non-apoptotic regulators in cell-death pathways.

The transcription factor ND-kB is a key regulator of a survival pathway in cancer cells and play a critical role in ongogenesis and tumor progression. Furthermore inhibition of ND-kB augments sensitivity to cancer therapies in various types of cancer. Mabuchi et al. recently linked NF-kB inhibition to an increase in cisplatin efficacy in in vitro and in vivo ovarian cancer models. Since NF-kB signaling is a focus of intense research in novel drug discovery, a better understanding of how NF-kB signaling is regulated would provide novel therapeutic targets to overcome drug resistance. In one of the genes identified, PLJ20241 showed higher expression in early recurrence in our study, and is a nuclear factor-B (NF-kB)-activating protein that was also identified as a gene that activates NF-kB and MAPK in a large-scale experimental study.

To further investigate the role of putative NF-kB activator in ovarian cancer chemoresistance we have generated expression constructs of FLJ20241 in both prokaryotic (PGEX) and Eukaryotic expression (pcDNA3.1) vectors. Also using the purified PLJ20241-GST fusion protein we have generated polyclonal antibodies to both the N- and C-terminal region of this protein. Using these antibodies, we have shown higher levels of FLH20231 expression in cisplatin resistant cell lines A2780/cp70 and A2780/C200 compared to the parental !2780 cisplatin sensitive line.

Based on the above results, we hypothesize that FLJ20241 could b used as a marker to predict chemoresistant disease. To further evaluate the biological effects of FLJ20241, we propose the following:

  1. To evaluate the effect of FLJ20241 expression on ovarian cancer resistance in vitro.
  2. To correlate the stable expression of FLJ20241 to chemoresponse, disease-free survival and overall survival in patient samples.
  3. To identify interacting proteins and initially map the pathways mediated by FLJ20241.

Investigation of the Contribution of the Fanconi Anemia Pathway to Ovarian Cancer Treatment

Ovarian cancer is the leading cause of death in women with gynecological tumors. Studies have been conducted on ovarian cancer treatment for over three decades, but no significant improvements have been made in the absolute cure rate of advanced-stage ovarian cancer. DNA-crosslinking agents, including Cisplatin/Carboplatin, have been extensively used in the chemotherapy for ovarian cancer. However, ovarian cancer patients have been found to develop resistance to these agents. The mechanisms underlying this resistance are not fully understood. It is known that the Fanconi Anemia (FA) pathway has been involved in the repair processes of DNA damage caused especially by these agents. This pathway is constituted by a group of FA proteins, and monoubiquitinated FANCD2 is a measure of this pathway activation. My preliminary studies and those of others showed that the impaired FA pathway conferred the cell sensitivity to DNA crosslinking agents. This let us hypothesize that the sensitivity/resistance of ovarian tumor to Cisplatin/Carboplatin may be also relevant to the integrity of FA pathway.

To test this hypothesis, we will pursue studies with two aims:

  1. To investigate the platinum sensitivity of ovarian cancer cell xenografts with different status of FA pathway in mice.
  2. To reveal the potential association between the status of FA pathway and outcomes of ovarian cancer patients treated with Cisplatin/Carboplatin.

The experiments in aim 1 will be conducted mainly through using a bioluminescent system to monitor the growth of ovarian cancer cell xenografts in mice treated with Cisplatin. The xenografts will be established from the ovarian cancer cells with an intact FA pathway and an impaired FA pathway, respectively. Thus the different outcomes of treatment directly result from the different status of the FA pathway. The experiments in aim 2 will be performed through detecting the status of FA pathway in ovarian cancer specimens available in the Mayo Clinic Ovarian Cancer Resources. This will give an indicative association between the FA pathway and the outcomes of chemo-resistance in ovarian cancer patients.

The results of this proposal will provide first-hand information regarding the implications of FA pathway in ovarian cancer treatment. The expected results will be in favor of manipulating the status of FA pathway during chemotherapy for increasing tumor sensitivity, and will suggest utilizing other chemotherapeutic agents if a given ovarian tumor harbors an intact FA pathway. Both will presumably be useful tools to improve the survival rate of patients with ovarian cancer.

Modeling Ovarian Cancer TCEAL-deficient Mice

Ovarian cancer etiology is not well understood because of the complexity of genetic alterations and the specific roles these alterations play in ovarian cancer initiation, progression and the development of chemoresistant disease. A better understanding of the earliest genetic changes that lead to a transformed phenotype may contribute to deciphering the underlying etiology of ovarian cancer. In contrast to other tumor types, efforts to model human ovarian cancer were delayed due to a lack of understanding of the disease process.

However, over the past five years a few genetically engineered mouse models of ovarian cancer have been generated. While each of these models has unique strengths, relevance of each model to human ovarian cancer is limited by the prevalence of these genetic alterations in ovarian cancer. The utility of some of these models is further limited by low frequency of tumor induction. These results suggest that additional genetic alterations are necessary for induction of ovarian cancer.

Recently, we have identified a novel tumor suppressor, TCEAL7, to be down-regulated in 90% of human ovarian cancer cell lines and human primary ovarian cancer. TCEAL7 is down-regulated in ovarian cancer of different histology, indicating that it may represent a common genetic alteration in the etiology of ovarian cancer. Specifically, down-regulation of TCEAL7 in the background of compromised p53 and Rb function was sufficient to transform nonmalignant human surface epithelial cells. This is the very first report of a novel tumor suppressor gene with the ability to transform nonmalignant ovarian surface epithelial cells in the absence of activated Ras and small t antigen. More importantly, since the expression of TCEAL7 is lost in 90% of ovarian cancer cell lines and primary ovarian tumors, we surmise that the loss of TCEAL7 expression could represent one of the earliest genetic alterations associated with the etiology of ovarian cancer.

Therefore, we hypothesize that TCEAL7 down-regulation is an initiating event in the transformation of normal ovarian surface epithelium concurrent with p53 inactivation. To test the role of TCEAL7 in ovarian cancer, we propose the following:

  1. To generate a TCEAL7 knockout muse model and investigate ovarian surface epithelium homeostasis.
  2. To investigate the cooperation between loss of function of TCEAL7 and p53 in the development of an ovarian cancer model.

2005 Ovarian Cancer Research Grant Funding

2005_OV_Grants

Folate-Receptor Targeted Measles Virotherapy for Ovarian Cancer

A Phase I clinical trial using live attenuated measles virus as a novel therapeutic for recurrent ovarian cancer is in progress at Mayo Clinic Rochester. This study will develop a new generation of targeted measles virus that exclusively infects ovarian cancer cells, thereby enhancing anti-tumor efficacy and minimizing toxicity.

Our goal is to develop replication-competent attenuated measles virus (MV) for targeted therapy of ovarian cancer (virotherapy). Attenuated MV is potently oncolytic for ovarian cancer cells and human tumor xenografts in mice. The virus uses its hemagglutinin (H) attachment protein to bind to CD46 receptors on ovarian cancer cells and enters cells using its fusion (F) protein. The virus then exploits the host cell machinery to express the viral proteins and for production of viral progeny. Subsequently, expression of MV-H and F proteins on surfaces of infected cancer cells results in extensive fusion with CD46 positive neighboring cells, leading to tumor cell destruction.

Encouraged by the promising anti-tumor activity of MV, we are currently testing intraperitoneal delivery of a recombinant MV (MV-CEA) in a phase I clinical trial for patients with recurrent ovarian cancer. The MV-CEA virus has been genetically engineered to express an inert soluble marker peptide to facilitate noninvasive monitoring of the profile of viral gene expression in patients.

Ovarian cancer is highly amenable to intraperitoneal measles virotherapy because the cancer is localized mainly in the peritoneal cavity. Direct delivery of the virus into the peritoneal space maximizes contact with tumor cells and minimizes potential neutralization by anti-measles antibodies in the blood. We are now designing the next generation of MV with enhanced performance for clinical testing. We hypothesize that the therapeutic index (efficacy versus toxicity) of oncolytic MV will be enhanced by developing viruses that infect exclusively ovarian cancer cells.

To test this hypothesis, we will generate targeted measles viruses ablated for binding to its two native cellular receptors, CD46, that is ubiquitously expressed on all human cells (except for erythrocytes) and SLAM (signaling lymphocyte activation molecule), which is expressed on activated B cells, T cells and monocyts/macrophages. To confer specificity, we will redirect MV binding and cell fusion exclusively to the alpha-folate receptor (aFR) via display of an anti-folate receptor single-chain antibody (scFv). Alpha-folate receptor is a highly desirable target in ovarian cancer therapy as it is overexpressed in more than 90% of epithelial carcinomas and the Mov18 antibody from which the scFv is derived has been tested in various clinical trials.

Mullerian Inhibiting Substance Type II Receptor: A Novel Ovarian Cancer Target

A major obstacle in the management of ovarian cancer is the inability to specifically distinguish the ovarian cancer cells, which would allow targeted therapy. Treatment strategies rely on a therapeutic ratio derived primarily from subtle differences between most cancers and normal cells in the ability to respond to DNA damage caused by anti-cancer agents. Current imaging strategies rely on nonspecific characteristics to differentiate tumor from normal tissues: recent advances such as PET-scanning have improved for some malignancies. The identification of tumor-specific characteristics would greatly enhance our ability to target ovarian cancer for therapy and imaging.

The regulation controlling the growth of the male or female reproductive structures during embryologic development is complex and depends upon tissue specific regulation of receptors. MISIIR is a receptor specifically expressed in the progenitor tissues of the female reproductive tract during embryology. In a male fetus the testis expresses MIS, the ligand for MISII, and this signals for regression of the mullerian tissues. In the female fetus, MIS is not expressed and the structures persist and develop further. MISIIR remains expressed in adult tissues apparently in a tissue-specific manner. Expression of MISIIR in a mullerian-specific manner may represent a great opportunity to selectively target ovarian cancer. We hypothesize that MISIIR expression is specific to the female reproductive tract and therefore an attractive target for therapy. We propose:

  1. To determine the tissue pattern of expression of MISIIR in adult females
  2. To analyze the expression of MISIIR in a stage-and histology-specific manner in primary and recurrent ovarian cancers
  3. To determine the fetal and adult patterns of MISIIR expression in mouse to explore the suitability of such a model for future development of MISIIR
  4. To improve the specificity of viral-mediated therapy for ovarian cancer using MISIIR as a target

The information obtained from this Minnesota Ovarian Cancer Alliance grant will provide preliminary data to a) determine the feasibility of pursuing MISIIR as an ovarian-specific target, and b) support an independent NIH grant application.

Novel Effect of a Mutant Endostatin in Inhibiting Peritoneal Dissemination of Ovarian Cancer

Cancer patients are currently treated by surgery and chemotherapy. While Taxol and platinum based drugs are the front line treatment choices, newer combinations with drugs such as Gemcitabine, 17-AAG, proteosome inhibitors are being introduced to aggressively eliminate the tumor cells. The ultimate goal of these combination regimens is to selectively inhibit the growth of cancer cells, and if possible, minimize toxicity. By understanding the biology of cancer cells it is possible to rationally design novel drugs in the future. But we have almost reached the threshold of chemotherapy beyond which the quality of life of cancer patients will be compromised.

In this context, it is important to investigate alternate targets, especially outside the tumor cells to complement existing methods of treatment. Four potential targets are amenable for therapeutic intervention in addition to the tumor cells.

  1. Vascular endothelium that provides nutrients and oxygen to the growing tumor,
  2. Tumor infiltrating lymphocytes,
  3. Stromal cells and
  4. Extracellular matrix that provides the scaffold for tumor cells to attach and remodel. Extracellular matrix also sequesters growth factors and provides them to cells.

Our work focuses on the vascular endothelium since it is the lifeline for the tumor tissue. Each endothelial cell in a blood vessel supports the growth of about 500 cancer cells. Therefore, inhibition of angiogenesis should profoundly affect cancer growth.

We have recently identified and characterized a mutant endostatin which was genetically modified to target tumor vasculature. The mutant endostatin bound to not only the endothelium but also ovarian cancer cells. Other antiangiogenic molecules such as angiostatin did not bind ovarian cancer cells. Based on this observation, we investigated whether the mutant endostatin can be used to block peritoneal dissemination of ovarian cancer cells. Ovarian cancer cells when shed into the peritoneum needs to seek out preexisting vasculature and co-opt with the vessels for survival. Attachment of cancer cells to a proximal site near a blood vessel is the first step in metastasis. Seeding is followed by angiogenic response which results in tumor growth. Our preliminary studies show that the mutant endostatin can indeed block the attachment of ovarian cancer cells to the peritoneal wall and prevent peritoneal dissemination.

Ovarian cancer is a unique target for the mutant endostatin which can be used to prevent cell attachment to the peritoneal wall in addition to inhibiting angiogenesis. We would like to expand on this novel effect of mutant endostatin so that we can develop a secondary prevention strategy to reduce recurrence rate of ovarian cancer.

Genetic Variation in Inflammation Genes and Associations with Ovarian Cancer Risk and Prognosis

Although it is clear that inherited genetic susceptibility plays a role in ovarian cancer, much remains unknown about the etiology and progression of this deadly disease. One promising area of research is the role of inflammation pathways in ovarian cancer. It is thought that the NFk B pathway (nuclear factor of kappa light polypeptide gene enhancer in B-cells) is a central mediator between inflammatory and carcinogenic processes.

We hypothesize that inherited variation in genes inhibiting or activating NFk B may modify risk of ovarian cancer and may also play a role in prognosis. We intend to examine these hypotheses in a population study using a comprehensive approach for evaluation of inherited variation in ~ 90 NFk B-related genes. Over 320 women with ovarian cancer and 395 healthy controls have been previously recruited into an ongoing case-control study at the Mayo Clinic and have provided risk factor information and genomic DNA; clinical follow-up data is available on the vast majority of cases.

To evaluate variation in approximately 90 candidate genes, we will take advantage of the recently-announced, new generation of genetic maps to identify the single nucleotide polymorphisms (SNPs) which are most informative for identifying or “tagging” underlying genetic differences across individuals. We expect ~ 480 “tagSNPs” to be identified through a bioinformatics screen of publicly-available databases and in-house statistical analysis. We will utilize cutting-edge technologies at the Mayo Clinic Advanced Genomic Technology Center for efficient and robust genotyping of each of these tagSNPs in our case-control study population.

These genotyping data from 328 cases and 395 controls will be statistically analyzed to assess whether any particular tagSNP is associated with increased or decreased risk of ovarian cancer. We will also consider joint effects of multiple tagSNPs within each candidate gene in a haplotype-based analysis. Among cases only, these genotype data will also be used to assess whether any particular tagSNP, or tagSNPs combination, is associated with longer or shorter time to disease recurrence. Our hope is that this comprehensive, yet efficient, approach to analysis of genetic variation in the NFk B pathway will highlight particular genes worthy of follow-up with additional molecular and population studies.

To our knowledge, this project will be the first to apply the novel genetic tagSNP approach in a comprehensive pathway-specific manner in ovarian cancer research. We will capitalize upon a vast amount of existing resources including previously-recruited ovarian cancer cases and controls with attendant biospecimens, state-of-the-art laboratory technology, and an active, multidisciplinary team of basic, clinical, and population scientists with proven expertise relevant to the discovery of genetic components for ovarian cancer. Increased understanding of the genetics of this disease at the population level will hopefully lead to targeted, more effective, prevention and therapeutic strategies.

2004 Ovarian Cancer Research Grant Funding

2004_OV_Grants

Diagnostic Markers for Ovarian Cancer: Detection of Novel Cell Adhesion Proteins in Patients’ Specimens

The study focuses on cell adhesion molecules present on the surface of ovarian cancer cells that appear to be unique to ovarian carcinoma. Once shed, these molecules will be present in the ascites fluid of the patient or will make their way into the bloodstream of the women. The study hypothesizes that some of these molecules are specifically expressed by ovarian carcinoma cells, but not expressed by normal or cancerous tissues. It is these molecules, used alone or in conjunction with CA-125, that can be used to develop a sensitive and specific diagnosis assay for the early detection and recurrence of ovarian carcinoma.

Modulation of Growth Factor Signaling by Hsulf-1: Implications on Ovarian Cancer Cell Survival and Chemo-resistance

Hsulf-1, an amino acid polypeptide, is decreased to less than 20% of normal levels in ovarian cancers and is undetectable in clear cell cancers, a particularly resistant subtype. Its downregulation appears to represent a mechanism by which ovarian cancer cells enhance growth factor signaling and diminished apoptosis (programmed cell death). The proposed research will define the role of Hsulf-1 downregulation in ovarian cancer and initiate a search for agents that might reverse such effects. If successful, the experiments will characterize a novel mechanism of drug resistance in cancer.

The Regulation of the Pro-Apoptotic Bcl2 Family Bok in Ovarian Carcinoma

The proposed research is based on the hypothesis that Bcl-2 family protein, Bok, is important for the survival and death of ovarian cells and defects in its regulation may underlie both tumorigensis and the resistance to drugs associated with ovarian cancer. The primary objective of this study is to investigate the regulation of the Bok protein in ovarian cells and carcinomas.

Functional Significance of Insulin Receptor Isoform A Expression in Ovarian Epithelial Cancer

While many ongoing studies attempt to identify the genetic changes present in ovarian cancer cells, more effort is needed in determining the functional significance of these changes. This information is required in order to select the most promising targets for new antioneoplastic therapies. Both treatment efficacy and reduction of adverse side effects would be aided by selection of tumor-specific proteins as targets. Recent findings about the insulin receptor may provide a unique opportunity for tumor-specific therapies. InsR-A is expressed at high levels in malignant ovarian cells. This study seeks to define its role in ovarian cancer with the potential aim of developing a therapy to inhibit InsR-A signaling that may impact cell proliferation, survival, and transformation.

2003 Ovarian Cancer Research Grant Funding

2003_OV_Grants

Anti-angiogenic Gene Therapy of Ovarian Cancer

Antiogenic therapies will be very useful in preventing recurrence of ovarian cancer after surgical debulking and chemotherapy. Lack of side effects from angiogenesis inhibitors allows its long term use. The study’s specific goal is, therefore, to develop a gene therapy method to express angiogenesis inhibitors in the peritoneum so that recurrence of ovarian cancer can be prevented.

Improving the Treatment of Ovarian Cancer: Understanding the Role of Spheroids in the Spread of Ovarian Carcinoma

This study will identify cells adhesion proteins on spheroids that allow them to adhere to the mesothelial cell layer, a common site of secondary tumor growth in ovarian cancer. Individual cells will be examined to establish their ability to migrate out of spheroids as well as their ability to invade through extracellular matrix molecules.

Functional Significances of Loss of Pro-apoptotic Protein HtrA1 in Epithelial Ovarian Cancer: Implications of Platinum Resistance

This study will define the role of HtrA1 down regulation in the clinical behavior of ovarian cancers in response to platinum based therapy. If successful, these experiments will not only characterize a novel mechanism of drug resistance, but also provide a basis for exploring novel approaches towards reversal of platinum resistance in ovarian cancer.

Hunger Pain in Ovarian Cancer Patients with Malignant Bowel Obstruction

This study would be a therapeutic trail to test whether a nicotine oral vapor inhaler might play a role in palliating hunger pain for women with malignant bowel obstruction who cannot eat. It would also explore the role of hormones, ghrelin [a hunger enhancer] and leptin [a hunger suppressor] in hunger pain.

2002 Ovarian Cancer Research Grant Funding

2002_OV_Grants

Continuous Ambulatory Infusion of Docetaxel: Anti-Angiogenic Chemotherapy for the Treatment of Recurrent Ovarian Cancer

The central hypothesis of this study is that continuous infusion of very low dose chemotherapy can be administered without significant toxicity while inhibiting tumor growth and enhancing patient quality of life. In the Phase I study, women with recurrent epithelial ovarian cancer will receive continuous infusion of docetaxel through a micro-infusion medication system. It is postulated that inhibition of tumor growth will be more profound in a continuous infusion model than in the traditional maximum tolerated dose model.

PARK2, A Mutational Target in Ovarian Cancer

Emerging evidence from the Mayo Clinic Cancer Center suggests that the large common fragile site gene PARK2 may play a role in the phenotype and/or the development of ovarian epithelial cancer. The study will analyze PARK2 alterations to determine at what point during ovarian cancer development these alterations may occur. The study will also examine the relationship between alterations in PARK2 and the resulting clinical phenotype of the ovarian tumor specimens.

The Use of a Spiritual Intervention to Enhance Mood States, Spiritual Well-Being and Quality of Life in Women with Ovarian Cancer

This pilot study will examine the effect of a spiritual intervention – centering prayer, a silent focused style of contemplative prayer – on mood status, spiritual well being and quality of life in women with recurrent ovarian cancer.  Women receiving the intervention will be compared to a group of women receiving standard medical care on measures of mood, spiritual well-being and quality of live.  A spiritual intervention with proven effectiveness that is transportable would be a potentially valuable coping resource for many patients.

Evaluation of Risk Factors Leading to Ovarian Cancer Metastasis at Laparoscopic Surgical Wound Sites

Laparoscopic procedures have been introduced in the diagnosis and management of patients with known malignancy. Unfortunately, tumor metastases at the port-site have been documented in ovarian cancer. A specific aim, using laboratory mice, is to evaluate the presence of growth factors present in laparoscopic vs. laparotomy surgical wounds and to decrease tumor recurrence at laparoscopic wound sites.

2001 Ovarian Cancer Research Grant Funding

2001 Research Funding Total $91,000