SPOREs : Organ-Specific SPORE Programs : Pancreatic

UNIVERSITY OF NEBRASKA MEDICAL CENTER SPORE IN PANCREATIC CANCER

Michael A. Hollingsworth, Ph.D., Principal Investigator
Professor
University of Nebraska Medical Center
Eppley Institute for Research in Cancer and Allied Diseases
986805 Nebraska Medical Center
Omaha, NE 68198-6805
Tel: 402/559-8343
Fax: 402/559-3339

For more information on this specific SPORE’s institution, please visit http://www.unmc.edu/eppley/spore.

The Specialized Program of Research Excellence in Gastrointestinal (Pancreatic Cancer) will focus on translational studies that address basic and clinical issues of importance to improving the outcome of patients with pancreatic cancer. Specifically, the research projects in this program seek to: 1) develop and test novel diagnostic reagents and assays that will improve our ability to detect pancreatic cancer in its early stages; 2) develop and test novel therapeutic strategies including immunotherapy, chemotherapy, and chemoradiation therapy for patients with early and advanced pancreatic cancer; 3) undertake basic research studies in conjunction with clinical trials that will provide insight at the molecular level into the reasons for success and failure of the different strategies. The SPORE program is focused on 4 projects with the high potential translational impact, and there are 3 highly interactive cores that will continue to acquire, store, and make available a unique set of tissue samples, data (clinical, molecular genetic, biological, pathological), reagents and resources:

PROJECT 1: IMMUNOTHERAPY OF PANCREATIC ADENOCARCINOMA

Principal Investigators: Michael A. Hollingsworth, Ph.D. and Jean Grem, M.D.

We propose to further develop and test novel reagents that can be used for immunotherapy of human adenocarcinomas, particularly those of the pancreas. The reagents under development in this project include highly specific murine monoclonal antibodies to circulating tumor-associated antigens (TAA), which form immune complexes that are taken up by dendritic cells (DCs) and other antigen-presenting cells (APCs) and are efficiently presented to the immune system. As a result, humoral and cellular immune responses against TAA are activated. The fundamental hypothesis under investigation is that murine antibodies against circulating human tumor antigens will bind to those antigens when administered to patients, form immune complexes that will be bound to APCs either directly or subsequent to the development of human anti-mouse antibody (HAMA) responses that capture these complexes, and that antigen processing by the APCs will produce immune responses against the targeted antigen. We specifically hypothesize that the anti-MUC1 antibody BrevaRex® MAb-AR20.5, when combined with soluble and/or cell-bound MUC1 in patients, will induce humoral and cellular immune responses to MUC1 that will be protective against pancreatic cancer in patients with MUC1-expressing pancreatic and other tumors. The strategy has the unique capacity to provide a method of vaccinating each patient with their own tumor antigens through in vivo capture and presentation of circulating and cell associated tumor antigens. We will target the cell surface associated mucin MUC1 with BrevaRex® MAb-AR20.5, a murine IgG1κ specific for the tandem repeat region of MUC1, which should provide effective targets for cell mediated responses against the tumor cells that produced the circulating antigen. One important challenge of producing effective tumor vaccines is developing reagents that break immunological tolerance to tumor-associated antigens. For preclinical studies, will utilize an inbred mouse strain on the C57BL/6 background that expresses human MUC1 in the correct temporal and spatial pattern (MUC1.Tg), develops tolerance and is refractory to immunization with MUC1. This experimental model has enabled us to study the effect of endogenous expression of the MUC1 gene on the ability of mice to produce protective immune responses to tumors, and represents an improved model system for evaluating the efficacy of anti–MUC1 formulations in vivo within the context of existing tolerance. We have developed and investigated a model in which a murine pancreatic tumor (Panc02) syngeneic to C57BL/6 transfected with human MUC1 (Panc02.MUC1), can be transplanted subcutaneously and orthotopically. In the studies proposed here, we will evaluate the mechanism of action of BrevaRex® MAb-AR20.5 in the murine model, conduct preclinical studies to determine its mechanism of action, and investigate the utility of combining this therapy with other interventions in a clinical trial in humans with pancreatic cancer.

PROJECT 2: INHIBITORS OF N-CADHERIN IN THE TREATMENT OF PANCREATIC CANCER.

Principal Investigators: Keith Johnson, Ph.D., Margaret Wheelock, Ph.D., and Jean Grem, M.D.

Pancreatic adenocarcinomas are among the most fatal cancers because of their extensive invasion into surrounding tissues and metastasis to distant organs, even at an early stage of tumor progression. The poor prognosis of this malignancy also reflects a generally poor response to current therapies. Thus, a basic understanding of the biology of these tumors and the mechanisms that promote their invasion and metastasis will provide a basis for developing new methods for diagnosis and treatment.

Pancreatic adenocarcinomas are characterized by extensive deposition of extracellular matrix, which can have profound effects on cell behavior. We have preliminary studies showing that cells derived from pancreatic adenocarcinomas respond in vitro to exogenous collagen type I by undergoing a transformation from a non-motile epithelial cell to a highly motile and invasive mesenchymal cell. A hallmark of epithelial to mesenchymal transitions is increased expression of N-cadherin, a protein we and others have shown promotes tumor cell invasion. Of particular significance to the current proposal, N-cadherin is expressed by more than 50% of invasive pancreatic tumors.

Recent studies have shown that the N-cadherin antagonist, ADH-1, developed by Adherex Technologies, Inc. Durham, NC, inhibits the activity of N-cadherin in vitro and in vivo. In addition, our data and that from the literature show that ADH-1 induces apoptosis in an N-cadherin-dependent manner. We hypothesize that ADH-1 is capable of inhibiting tumor growth and metastasis in N-cadherin-expressing pancreatic cancer cells, and propose to test this hypothesis in N-cadherin-expressing pancreatic cancer cells in a mouse model of invasive pancreatic cancer. In addition, we propose to characterize the signaling pathways downstream of collagen I that promote up-regulation of N-cadherin and invasion, for the purpose of identifying potential inhibitors that could be used in combination therapy with ADH-1. Finally, we will test the efficacy of ADH-1 as a treatment for human pancreatic cancer in clinical trials.

PROJECT 3: BIOLOGICAL MARKER(S) IN THE DIAGNOSIS OF PANCREATIC CANCER

Principal Investigators: Surinder Batra, Ph.D., Randall Brand, M.D., and Aaron Sasson, M.D.

Pancreatic cancer (PC) is a disease of insidious progression and high lethality, with the majority of tumors having extended beyond the confines of the pancreas at the time of diagnosis. The early diagnosis of PC remains a challenging endeavor that will require multidisciplinary approaches, including imaging studies, tissue sampling, and analysis of body fluids from patients. There is no specific tumor marker available for diagnosing pancreatic cancer. The principal reason for this grim prognosis is our inability to diagnose the disease at an early, localized, and potentially curable stage. In an extensive study on the expression analysis of various mucin (MUC) genes, we demonstrated specific and differential expression of MUC4 in pancreatic adenocarcinomas as compared to the normal pancreas or chronic pancreatitis tissues.

The central hypothesis of this proposal is that the MUC4 is detected in peripheral blood mononuclear cells (PBMCs) and fine needle aspirates (FNAs) of pancreatic cancer patients but not of healthy individuals or those with non-neoplastic pancreatic diseases.

Three specific aims are proposed.

• Aim I will compare the expression of MUC4 by real-time RT-PCR in PBMCs of patients with pancreatic cancer, pre-malignant pancreatic diseases, pancreatitis and aged-matched healthy subjects. This aim will establish the sensitivity, specificity, accuracy and diagnostic potential of MUC4 mRNA in PBMCs. Additionally, it will determine whether the detection of MUC4 mRNA in blood correlates with tumor stage, tumor progression, recurrence or response to therapy.
• Aim II will identify the cell types in PBMCs that are positive for MUC4. A subset of patient’s PBMCs will be FACS-analyzed to investigate the nature of the cell type(s) that express MUC4 mRNA. We will investigate the possibilities that MUC4 in PBMCs is from: i) circulating pancreatic tumor cells; ii) circulating dendritic cells or macrophages that have acquired MUC4 mRNA through phagocytosis or pinocytosis of primary or metastatic tumor cells; or iii) PBMCs that have turned on the expression of MUC4.
• Aim III will investigate the expression of MUC4 and nanostructural cytological changes in fine needle aspirates (FNAs) from PC patients and attempt to distinguish them from chronic pancreatitis and other benign diseases using immunocytochemical and partial wave spectroscopic (PWS) analyses. MUC4 protein will be measured in FNAs and will be correlated with the sensitivity and specificity of diagnosis. In addition, partial wave spectroscopy (PWS) will be utilized to improve the ability of cytological examination on FNAs to distinguish PC cells from non-neoplastic cells. Associating PWS signatures with MUC4 expression will further assist in disease classification and planning/formulating management strategies.

Taken together, these aims will establish the utility of MUC4 in the diagnosis of pancreatic cancer

PROJECT 4: INHIBITORS OF TELOMERASE IN TREATMENT OF PANCREATIC ADENOCARCINOMA

Principal Investigators: Michel Ouellette, Ph.D., Jerry Shay, Ph.D., and Jean Grem, M.D.

Telomeres progressively shorten in almost all normal human tissues with increased age. In pre-neoplasia, telomeres are exceptionally short. Almost all human malignant tumors express telomerase to maintain these short telomeres. This activity is absent or is at lower levels in normal tissues. This has led to approaches for inhibiting telomerase as a target for cancer therapeutics. Cancer stem cells have been shown to be both telomerase expressing and containing short telomeres. This suggests that telomerase inhibitors are likely to target both the bulk of the more differentiated tumor cells as well as the stem cells that provide the unlimited growth of most advanced cancers. This project will conduct pre-clinical orthotopic xenograft animal experiments followed by clinical trials in patients with pancreatic cancer. The preclinical experiments will evaluate the in vivo effectiveness of a novel telomerase inhibitor just entering clinical trials, GRN163L, on human pancreatic cancer cells in mice bearing orthotopic tumors in the pancreas. GRN163L has enhanced stability and displays extremely specific and high-affinity binding to telomerase. These properties increase the chances of effectively inhibiting telomerase in cancer cells throughout the body with relatively low doses of the drug. We have already demonstrated that our lead compound, GRN163L, is an effective inhibitor of telomerase in several types of cancer cell lines. The proposed experiments will use bioluminescence imaging to follow human pancreatic tumor cell lines treated in vivo with GRN163L alone and in combination with chemotherapy. The goal is to monitor the spatio-temporal distribution of pancreatic tumor cells during the disease course from minimal residual disease to metastasis with and without GRN163L and combinations of chemotherapy. The experiments should unambiguously prove or disprove the link between the inhibition of telomerase and decreased proliferation of tumor cells in vivo. In this proposal, we will exploit this knowledge to conduct early stage clinical trials on pancreatic cancer patients with locally advanced, unresectable, or metastatic disease. For these patients, a standard treatment option is the combination of gemcitabine with erlotinib (www.nccn.org). In a phase I/II trial, prolonged exposure to GRN163L will be administered in combination with gemcitabine/erlotinib chemotherapy, and the safety and efficacy of GRN163L will be determined. The primary endpoints will be telomerase inhibition, survival and progression-free survival. From a subset of patients, pancreatic juice will be obtained to measure the impact of GRN163L on telomerase activity at the tumor site. From patients that agree to participate in an ongoing rapid autopsy program, measurements of telomere size in tumors will also be obtained at time of death. This trial is likely to the first clinical trials testing a telomerase inhibitor in patients with pancreatic cancer.

CORE A: ADMINISTRATION

Core Director: Michael A. Hollingsworth, Ph.D.
Co-Directors: Michael Brattain, Ph.D. and Jean Grem, M.D.

The administrative core will provide support for personnel involved in the administration, scientific oversight and coordination, communication, budgetary oversight, and clerical functions associated with the overall research program proposed in this SPORE application. The specific functions of the core include:

1. Monitoring progress of the translational research objectives of the SPORE program.
2. Facilitation of internal and external communication among SPORE investigators, and communicating reports of progress from SPORE projects with other SPORE programs, the NIH, and the scientific and public communities.
3. Coordination and scheduling of all external and internal SPORE meetings.
4. Administration of the Developmental Research Program.
5. Administration of the Career Development Program.
6. Providing support for management of financial accounts related to SPORE activities and providing fiscal oversight of SPORE Projects.

CORE B: PANCREAS TUMOR SPORE TISSUE BANK

Core Director: Julia Bridge, M.D.

All projects in this SPORE will use human specimens for translational research directed at reducing the incidence and mortality of pancreatic cancer. In order to provide the necessary specimens, a Pancreas Tumor SPORE Tissue Bank has been developed in cooperation with and under the auspices of the University of Nebraska Medical Center (UNMC) Tumor Bank. There are two components to this resource. The first component is a conventional tissue bank, which captures and stores all excess surgical material from patients seen at UNMC and affiliated institutions, related to pancreatic and GI malignancies. This tissue bank also captures and stores samples related to clinical trials that are ongoing as part of SPORE activities. The second component is an organ harvest/rapid autopsy program in which patients who die with pancreatic cancer donate their organs for research purposes. This program captures and processes entire internal organs from these patients, including all available samples of primary tumor and metastases. The guidelines and protocols for collecting specimens established by the UNMC Tumor Bank and the Institutional Review Board are followed for the SPORE proposal. This core facility stores normal, benign (i.e. acute and/or chronic fibrosing pancreatitis) and malignant pancreatic tissues (including both primary and metastatic pancreatic carcinomas) and peripheral blood lymphocytes, plasma and serum from patients with pancreatic malignancies. The Bank also coordinates collection and storage of pancreatic ductal secretions and peritoneal washings. Cytogenetic analysis is performed on malignant lesions when possible. The core includes a mechanism for database management and specimen distribution. A uniform system of prioritization of requested materials has been defined and used by the Pancreas Tumor SPORE Tissue Bank Oversight Committee. This core facility is intended to benefit the specific research activities of the SPORE, as well as the research activities of other scientists within and outside of UNMC who are concentrating on translational research issues. Additionally, tissues are available for distribution through NCI supported tissue networks in national prioritization. Only specimens obtained from clinically indicated surgeries after all other diagnostic procedures have been performed are submitted to the Pancreas Tumor SPORE Tissue Bank for translational research. The specimens would otherwise be discarded or disposed of. Eligible patients have the opportunity to participate by submitting written informed consent. There is no risk to the patient or compromise to the patient's care, since all of the procedures performed would be performed for diagnostic reasons regardless of the SPORE. Members of the pathology department and the clinical departments are participants in the individual research projects and thus, also contribute to the Core for maximal and effective accumulation of satisfactory specimens.

CORE C: BIOSTATISTICS

Core Director: Jane Meza, Ph.D.

The Biostatistics Core will develop statistical designs, develop statistical analysis plans, perform data acquisition, develop databases, archive data, develop protocol specific clinical research forms, perform data monitoring for clinical trials involving human subjects, perform data analysis and interpretation and aid in the development of manuscripts, abstracts and presentations. The specific aims of the Biostatistics Core are:

• Aim 1: Collaborate with SPORE investigators on study design to develop projects with appropriate statistical operating characteristics.
• Aim 2: Provide a statistical analysis plan for each SPORE project.
• Aim 3: Perform statistical analyses of project results as specified in the statistical analysis plan and collaborate with SPORE investigators on the interpretation and reporting of research findings.
• Aim 3: Monitor the conduct of SPORE clinical research projects to ensure patient safety and the scientific integrity of the projects.
• Aim 4: Collaborate with SPORE investigators on interpretation of data analysis results and aid in the development of manuscripts, abstracts and presentations.
• The Biostatistics Core will provide investigators with the biostatistical and computing expertise that is essential to the design of laboratory-based and clinical studies because it ensures the efficient use of financial, animal and patient resources while minimizing the likelihood of false conclusions. The projects of this SPORE will generate complex data that will require substantial biostatistical and computing input to be effectively analyzed and presented.

CAREER DEVELOPMENT PROGRAM

Directors: Michael A. Hollingsworth, Ph.D., Michael Brattain, Ph.D. and Jean Grem, M.D.

Career Development Awards will be used to support the development of careers in translational pancreatic cancer research by individuals from one of the following categories:

1. Clinical fellows in medical oncology, gastroenterology, pathology, or surgery;
2. Junior faculty members at the instructor or assistant professor level, either clinical faculty who seek training and an opportunity to establish a career that includes basic science research, or basic science faculty who want additional training or an opportunity to undertake translational research;
3. Senior postdoctoral fellows who are at the point of entering the academic ranks and establishing independent research programs;
4. Established clinical faculty who require additional training in basic or translational research in order to establish a translational research program.

We will support one new candidate in each year of the grant for two years. Individuals in this program are required to commit substantial effort to SPORE-related translational research. Funds can be used for salaries or any other aspect of development (research costs) that are deemed necessary for the candidate to succeed in establishing a translational pancreatic cancer research program. The candidate is required to submit an NIH G or R series grant application during the second year of the award that is primarily focused on pancreatic cancer research.

DEVELOPMENTAL RESEARCH PROGRAM

Principal Investigator: Michael A. Hollingsworth, Ph.D.
Directors: Michael Brattain, Ph.D. and Jean Grem, M.D.

The principal goal of the Developmental Research Program is to fund promising early stage projects that address important translational objectives in prevention, early detection, and therapy of pancreatic carcinoma. Our principal intent for the Developmental Research Program is to bring novel translational research projects to the SPORE program. Key elements that determine priority in this program are innovation, novelty, and potential for success in translational pancreatic cancer research. All Developmental Funds will be awarded by competition based on submission of a NIH-style pilot project application. There is a requirement for clear evidence that the proposed project shows potential of developing into a larger research project that will involve human intervention for pancreatic cancer (diagnostic or therapeutic). Proposals are received and processed by the Administrative Core. Once a year, a request for proposals (RFP) is developed and distributed by the Principal Investigator. Two types of applications will be solicited: applications from single investigators for focused projects, or larger collaborative translational projects involving two or more investigators. Applications are reviewed by a study section comprised of the Internal Advisory Board, members of the SPORE program who are not in conflict, selected scientists and clinicians from the UNMC Eppley Cancer Center. Final selection of the funded applications is made by the External Advisory Board, who acts as an Advisory Council, in consultation with Dr. Brattain and Dr. Grem, who are the administrative leaders of the Developmental Research Program. Funded projects must have a set of quantifiable milestones for achievement that represent acceptable progress by the end of the first year.

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