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MAYO CLINIC SPORE IN PANCREATIC CANCER
PRINCIPAL INVESTIGATOR: GLORIA M. PETERSEN, PH.D. Mayo Clinic has had a long tradition of research in pancreatic cancer, and the SPORE strengthens and extends this tradition. New avenues of investigation will include gene and environment interaction, and roles of two very interesting genes that have been implicated in pancreatic cancer. These studies should lead to new insights that will translate into strategies for prevention, early detection, and treatment. The SPORE includes four research projects supported by three scientific core resources, and an Administrative Core. Several innovative translational pilot projects will be awarded annually through a Developmental Research Program with matching funds from the institution and a Career Development Program will support one junior faculty member each year. Project 1: Molecular Epidemiology of Pancreatic Cancer Molecular epidemiologic risk factors for pancreatic cancer have not been well studied. Even carefully designed "population-based" studies suffer from a limited accrual of biological samples because of the late diagnosis and rapid demise of pancreatic cancer patients. Clinic based studies can overcome this problem because of the rapid access to patients around time of diagnosis. We propose a clinic-based case-control study on the molecular epidemiology of pancreatic cancer, including 1200 ultra-rapidly recruited pancreatic cancer cases and 1200 unrelated primary care clinic controls. Controls will be frequency matched to cases on age, race, gender, and state/region of residence. Questionnaires will be used to collect data on hypothesized exposures, including smoking, non-steroidal anti-inflammatory drugs (NSAIDs), dietary carcinogens from meat and meat preparation, and family history. Blood samples will be collected on all cases and controls. Candidate gene polymorphisms to be studied are genes related to smoking (NAT2, GSTM1, GSTT1, NQO1, CYP1A1, and CYP2E1). Genomic control genes, not hypothesized to be associated with pancreatic cancer, will also be examined in order to evaluate and control for population stratification. Other sources of bias related to recruitment intervals and referral will be evaluated. Risk of pancreatic adenocarcinoma associated with candidate genes, environmental exposures, and gene-environment interactions will be estimated. Increased risk associated with smoking and family history of pancreatic cancer will be confirmed, and odds ratios for pancreatic cancer risk with use of NSAIDS and potential dietary carcinogens from meat/preparation will be estimated. Odds ratios for each candidate gene with smoking and family history will also be estimated, with a secondary analysis of the odds ratios for each candidate gene with exposure to dietary carcinogens from meat/preparation. This study will have significant translational impact because it will identify risk factors for pancreatic cancer that should be meaningful and eliminate spurious associations that were based on inadequate samples. By clarifying the genes and risk factors that are relevant to study in pancreatic cancer, this study will inform intervention strategies and basic research by suggesting potential genetic and carcinogenic mechanisms for this cancer. Project 2: SDF-1a/CXCR4-EGFR Interactions in Pancreatic Cancer Morbidity and mortality from pancreatic cancer is strongly associated with the metastasis, or migration, of pancreatic cancer cells. Unfortunately, lack of knowledge about the basic molecular mechanisms responsible for pancreatic cancer metastasis and progression significantly impairs the development of more effective treatments for this painful and rapidly fatal disease. The chemokine, SDF-1alpha, and its receptor, CXCR4, normally function to promote the in vivo motility of many cell types, and SDF-1alpha/CXCR4 signal transduction has recently emerged as a promising molecular target for modulating the growth and metastasis of several epithelial cancers. Our preliminary results establish a likely role for SDF-1alpha/CXCR4 signal transduction via the Epidermal Growth Factor Receptor (EGFR) in pancreatic cancer cells triggering simultaneously pancreatic cancer cell motility and a paracrine angiogenic response, both phenomena which are critical for the development of metastasis. Here, we propose to investigate the role(s) of SDF-1alpha/CXCR4 signaling via the EGFR in these phenomena using state-of-the-art in vitro and in vivo approaches provided by a multidisciplinary team of basic and clinical cancer investigators. Moreover, we propose translational experiments that evaluate the potential of blocking the CXCR4/SDF-1alpha pathway alone or in combination with the EGFR to inhibit metastasis formation. Our central hypothesis is that the SDF-1alpha-CXCR4 pathway interacts with EGFR signaling to trigger pancreatic cancer cell motility (cellular response) and angiogenesis (paracrine response) and its blockage interferes with tumor growth and metastasis. Considering that inhibitors for several components of this signaling pathway, including inhibitors of CXCR4 and the EGFR themselves, are at different stages in clinical trials, the proposed study has significant translational potential. Specifically, the proposed studies have the potential to provide a novel initial justification for the clinical testing of these drugs in patients afflicted with pancreatic cancer. Project 3: Mechanism of VAV1-Mediated Pancreatic Cancer Cell Growth Pancreatic ductular adenocarcinoma is the most common pancreatic tumor accounting for more than 90% of all pancreatic cancers and ranks fifth as a cause of death by cancer in the USA. Therefore, efforts to understand the molecular mechanisms underlying the development of pancreatic cancer may lead to preventative and treatment strategies to improve the outcome of the disease. Although the underlying etiology and pathophysiology of pancreatic ductal cancer is poorly understood, there is an increasing body of published work, and preliminary data presented in this application suggesting that signaling pathways that control cell proliferation, differentiation, and apoptosis are dysregulated in pancreatic cancer. Our identification of the hematopoietically-expressed proto-oncogene Vav1 in greater than 50% of primary pancreatic ductular adenocarcinomas together with the preliminary data described in the proposal indicate that Vav1 is involved in the regulation of pancreatic tumoral cell growth. The mechanistic experiments outlined in this proposal will test the central hypothesis that Vav1 expression in certain pancreatic cancers can potentially control tumor cell growth. We hypothesize that: (a) Vav1 stimulates the growth of pancreatic cells in vitro; (b) Vav1 regulates the cell cycle in pancreatic cancer by up-regulating the expression of G 1 phase cyclins; (c) Vav1 regulates the clclin D1 promoter in pancreatic tumoral cell lines through the activation of Rho family GTP-binding proteins; (d) Vav1 synergizes with oncogenic Ras in the regulation of cyclin D1 activity; (e) Vav1 regulates cycline D1 activity, at least in part through its regulation of the transcription factor NF- k B, and (f) Vav1 expression is acquired during tumor development and influences the pathobiological behavior of the disease. In order to test these hypotheses we will (1) Determine the role of Vav1 in pancreatic tumoral growth; (2) Characterize Vav1-mediated signaling cascades that control cell growth in pancreatic cancer; (3) Implement Vav1-based translational efforts for pancreatic cancer. Together these studies will provide an experimental basis for understanding the molecular events that are involved in the regulation of pancreatic tumor cell growth by Vav1, and will, in a broader context, advance our understanding of fundamental processes involved in cell surface receptor-mediated signaling cascades controlling cell growth in epithelial tumors. In addition, the translational efforts put-forth in this proposal will provide the experimental basis for the development of diagnostic and therapeutic options for the treatment of this deadly disease. Project 4: Characterization of the Role of BRCA2 in Pancreatic Cancer Mutations in the BRCA2 gene account for up to 17% of familial pancreatic cancer families and play a role in as many as 10% of pancreatic ductal adenocarcinomas in the Ashkenazi Jewish population. While BRCA2 appears to have a significant role in the development of pancreatic cancer, the specific functions and functional domains of the modular BRCA2 protein that must be inactivated in order for tumor formation to occur have not yet been determined. Similarly, the interaction of BRCA2 with pathways that are regulated by well-characterized pancreatic tumor suppressors and oncogenes remain to be established. In addition, even though BRCA2 mutant pancreatic tumors appear to be significantly different from sporadic tumors at the genetic and molecular level, no specific therapies for treatment of these tumors have yet been identified. With these questions in mind, we propose a comprehensive study of the role of BRCA2 in pancreatic cancer. Initially, in Aim #1 we will determine the effect of disruption of independent functional domains and regions of conserved sequence of BRCA2 on pancreatic cell lines in an effort to identify those domains that mediate normal function of BRCA2. This study will inform about the mechanism of action of BRCA2 and will also allow us to identify BRCA2 missense mutations from cancer patients that contribute to cancer development. In Aim #2, we will use knockout mouse models to evaluate whether disruption of the p53 and K-Ras signaling pathways, that have been found to be mutated in BRCA2 mutant cell lines and tumors, co-operate with BRCA2 inactivation to promote pancreatic tumor formation. In Aim #3, we propose a preclinical study to evaluate the efficacy of mitomycin C and cisplatin as treatments for BRCA2 mutant pancreatic tumors. As BRCA2 mutant cells repair intra-strand DNA crosslinks through error prone pathways, we hypothesize that agents such as mitomycin C and cisplatin will induce high levels of DNA damage and apoptosis in these cells and will be particularly effective agents for treatment of BRCA2 mutant pancreatic tumors. We will use orthotopic pancreatic tumor xenografts of BRCA2 mutant cell lines and tumors for these experiments. Taken together, this comprehensive evaluation of the role of BRCA2 in pancreatic cancer will improve our understanding of familial pancreatic cancer, the mechanisms behind pancreatic tumor development, and the most appropriate therapies for this form of familial pancreatic cancer. Core 1: Administrative Core The Administrative Core will provide organizational support for the leadership of the SPORE and facilitate communication among the component activities of the SPORE and with non-SPORE parties. The Administrative Core will have the following functions. It will: 1) Provide leadership and coordination between the Research Projects and Cores of the SPORE; 2) Assure ongoing integration and participation of the Pancreatic Cancer SPORE in the activities of Mayo Clinic Cancer Center; 3) Organize quarterly meetings of the SPORE investigators; 4) Organize yearly meetings of the External Advisory Committee; 5) Organize meetings of the SPORE Scientific Advisory Committee; 6) Organize meetings of the SPORE Executive Committee; 7) Provide administrative support to the Developmental Research Directors; 8) Provide administrative support to the Career Development Program Directors; 9) Facilitate investigator trips to the annual SPORE meetings in Washington D.C. ; 10) Organize a debriefing and planning session following the return from the annual SPORE meeting in Washington D.C. ; 11) Prepare the yearly non-competing SPORE application; 12) Serve as a liaison between the Mayo Clinic SPORE and the NCI SPORE Program, other SPOREs, and collateral resources, such as Rochester Epidemiology Project, PACGENE, PANCAN, and the North Central Cancer Treatment Group; 13) Create and maintain a Mayo Pancreatic Cancer SPORE website that will be useful to investigators inside and outside the SPORE, as well as patients; 14) Coordinate information and communication about SPORE-related research developments to and among the Mayo Clinic SPORE investigators, to the scientific community at large, and to the public. The Core leader, Dr. Petersen, will direct this Core so that maximum potential for translational objectives can be achieved. Core 2: Patient Registry and Biospecimens Core The goal of this core is to provide investigators in the SPORE high quality patient data, DNA, RNA, serum, and pancreas tissues from consented pancreatic cancer patients and to make these resources available for future studies. The activities of the CORE will be conducted in a way that does not compromise patient confidentiality, yet will be as comprehensive as possible (within its scope) in the materials that are provided. The acquisition of human tissue and subsequent cellular/molecular analysis of that tissue within the context of patient data are key to many laboratory-based studies of cancer. The Mayo Clinic has a strong tradition of tissue acquisition and patient data records. Paraffin embedded tissues, histological slides, and associated patient charts from surgeries performed since the first decade of the 1900's are maintained in Mayo’s Tissue Registry and the Mayo Archives. Today, the Tissue Acquisition and Cellular/Molecular Analysis (TACMA) Shared Resource of the Mayo Clinic Cancer Center provides normal and neoplastic human tissues for cancer research at Mayo, and is a resource of expertise, collaborative support, and service for pathology, immunohistochemistry, in situ hybridization, laser capture microdissection, tissue microarray preparation, and RT-PCR. The Patient Registry and Biospecimens Core of this SPORE will be integrated with the existing TACMA Shared Resource in order to provide a coordinated, centralized, dedicated program for procuring, processing, and assessing biospecimens and patient data from pancreatic cancer patients. To achieve our goals, we have assembled a clinical research team of experts in pancreatic cancer, consisting of Drs. S. Chari (gastroenterologist), M. Farnell (GI surgeon), S. Alberts (medical oncologist), G. Kim (oncologist, Jacksonville), G. Petersen (epidemiologist), L. Miller (GI research, Scottsdale), and M. Sarr (GI surgeon), to support the activities of the Patient Registry and Biospecimens Core under the combined leadership of Dr. W. Lingle, Co-Director of the TACMA Shared Resource of the Mayo Clinic Cancer Center, and Dr. T. Smyrk, anatomic pathologist. The Patient Registry and Biospecimens Core activities will be closely coordinated with the Biostatistics and Bioinformatics Core, for data management and analyses. We will install new infrastructures at Mayo Clinic Jacksonville to recruit pancreatic cancer patients and their biospecimens into a unified registry under this Core. Core 3: Proteomics Core The Proteomics Core (PC) has been an important shared resource of the Mayo Clinic Cancer Center (MCCC) since 1992. In the past decade, the PC has continued its rapid growth and expansion of core services, and has been an essential resource to Mayo investigators through its support of a broad diversity of projects related to cancer including pancreatic cancer. Usage of this core has grown substantially from 21 members in 1992, to more than 74 current members in 2004. The PC provides protein analysis and identification by MALDI-TOF mass spectrometry and MS/MS methods. In addition, the PC will develop an integrated biomarker analysis and discovery platform to support the longer-term goals of several pancreatic SPORE members; this platform is based on Fourier Transform Ion Cyclotron Resonance (FT-ICR) mass spectrometry. The PC resource will also continue to provide basic services in protein chemistry, which include solid phase peptide synthesis, protein purification, Edman chemical protein sequencing, and 2-D polyacrylamide gel electrophoresis. Usage of these services by pancreatic SPORE members will be given high priority compared to other requests of the core. A list of current services provided to members of the pancreatic SPORE include: (1) Solid phase peptide synthesis by Fmoc methods; (2) Edman N-terminal protein sequencing; (3) Protein purification by reverse phase HPLC and low pressure FPLC; (4) Computer-aided analysis of protein sequences for synthetic peptide design; (5) 1-D and 2-D PAGE separation of complex protein mixtures; (6) MALDI-TOF mass spectrometry analysis of proteins and peptides; and (7) Protein identification by Ion-Trap LC-MS/MS mass spectrometry. Core 4: Biostatistics and Bioinformatics Core The Biostatistics Core provides statistical collaboration and data management support for each of the SPORE projects, the developmental projects, and the Cores. Each of the projects presented in this application reflects input from members of the Biostatistics Core on study design and analysis plan. The Biostatistics Core will provide statistical support across many different fields, including epidemiological studies, basic sciences including gene array, and clinical trials. This comprehensive nature of the Biostatistics Core assures each SPORE investigator access to statistical expertise that includes collaborative development of study designs and analysis plans, state of the art data analysis and interpretation, data management resources, and abstract and manuscript preparation. The Biostatistics Core also provides a mechanism for the management and integration of both existing and newly collected data through consistent and compatible data handling. Areas of support include database development, data form development and processing, quality control, data collection and entry, and data archiving. This Core complements and assists the efforts of other Cores such as the Patient Registry and Biospecimens and Animal Models and Xenograft Cores with superior data management and experience with the Pancreatic Cancer Registry. The Biostatistics Core builds upon the innovative and time-tested procedures and systems developed by one of the largest statistical groups in the country whose members have collaborated on more than 8,000 clinical and basic science research studies since 1966. Developmental Research Program The Mayo Clinic SPORE in Pancreatic Cancer Research will make every effort to maximize the number of innovative and high-quality projects in the Developmental Research Program. The goal of this Program is to support innovative, scientifically sound research projects from which findings can be translated into clinically relevant applications that will impact screening, diagnosis, and management of pancreatic cancer. This Program will: (1) encourage and solicit innovative translationally-relevant laboratory, population and clinical study proposals; (2) encourage and support interdisciplinary collaboration in translational research in pancreatic cancer; and (3) generate new hypotheses that can be tested in larger-scale research projects or clinical trials that can impact pancreatic cancer. The availability of this support provides a stimulus for creativity in the research community, a vehicle for encouraging the interaction of basic scientists and translational investigators, and an opportunity for expanding the research spectrum of the SPORE by pursuing new leads based on discoveries and/or opportunities that arise. The Developmental Research Program will provide pilot funds for one year to three to five projects. A process is in place to call for applications and to formally peer review submissions utilizing the expertise of the Scientific Advisory Committee and other experienced investigators. Criteria will be based upon scientific merit, originality, qualifications of the key personnel and interactions, and translational potential. It is anticipated that support of developmental research projects will result in the generation of new hypotheses that can potentially be addressed in existing SPORE-sponsored projects, or through peer reviewed external grant support. It is the intent of the SPORE leadership to encourage and help the investigators to use the data generated by these projects to design either R01 or Program Project Grant proposals before the end of the five years funding period covered by this grant. Career Development Program The Mayo Clinic SPORE in Pancreatic Cancer will focus its Career Development Program on mentoring talented junior faculty who can develop independent research programs in translational pancreatic cancer research. One of the starkest realities facing the contemporary research community is the paucity of investigators involved in translational pancreatic cancer research. We recognize the importance of training and inspiring individuals who are at an important junction of committing to a career research program. At Mayo Clinic, we are uniquely qualified to make our proposed Career Development Program for junior faculty the most fruitful strategy to address the problem. These strategies include our existing core of committed and experienced pancreatic cancer researchers, combined with a highly productive team of investigators in other areas of cancer research who can serve as mentors. Mayo Clinic has, by its seamless blend of patient care and basic and applied research facilities, an environment conducive to translational research. There exists a continuous pool of outstanding scientists and clinicians (including talented female and minority investigators) who are early in their careers and who need exactly an impetus such as that offered by our SPORE's proposed Career Development Program to engage in translational research with a focus on pancreatic cancer. The Career Development Program is designed to increase the translational investigator base in the Mayo Clinic SPORE in Pancreatic Cancer by identifying and fostering the early careers of junior investigators. In this SPORE, we have adopted the concept of interdisciplinary mentoring teams for the junior faculty, because there are talented senior scientists who can co-mentor in their area of expertise, along with those mentors whose expertise is in pancreatic cancer. We propose formal mechanisms for recruiting, selecting and evaluating awardees, and will ensure that awardees are integrated into the SPORE research environment. In all cases, we expect that recipients in the career development program would build upon the resources allocated to them to develop independent funding in pancreatic cancer research. |
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