Developing a Screen for Novel Therapies with Reprogrammed Pancreatic Cancer Cells

描述（由申请人提供）：我们提案的目标是开发一种创新的检测方法，使用来自原发性胰腺导管腺癌（pda）的重编程干细胞来筛选新的个性化治疗。我们的长期目标是发现有效的抗癌药物并治愈PDA。我们的申请与NIH胰腺癌试点项目倡议高度相关，因为我们建议：1)开发一种新的基于细胞的检测方法，使用直接来自患者肿瘤的重编程细胞，2)利用重编程细胞筛选有效的个性化治疗方法，使用独特的临床就绪药物库。我们创新的检测方法是基于最近的发现，即肿瘤和非恶性细胞从患者活组织检查中获得，可以通过在类似胚胎干细胞的条件下培养细胞来扩增。这种策略在正常上皮细胞或癌细胞中诱导成体干细胞表型。具体来说，Rho激酶抑制剂（Y-27632）与小鼠喂养细胞的结合将上皮细胞重编程为干细胞样细胞

DESCRIPTION (provided by applicant): The goal of our proposal is to develop an innovative assay using reprogrammed, stem-like cells derived from primary pancreatic ductal adenocarcinomas (PDAs) to screen for new, personalized therapy. Our long- term goal is to discover effective anticancer drugs and cure PDA. Our application is highly relevant to the NIH Pancreatic Cancer Pilot Project initiative because we propose to: 1) Develop a novel cell-based assay with reprogrammed cells derived directly from patient tumors, and, 2) Harness the reprogrammed cells to screen for effective, personalized therapy with a unique library of clinic ready agents. Our innovative assay is based on the recent discovery that tumor and nonmalignant cells from patient biopsies can be expanded in culture by growing the cells under conditions similar to those used for embryonic stem cells. This strategy induces an adult stem cell phenotype in normal epithelial or carcinoma cells. Specifically, the combination of Rho kinase inhibitor (Y-27632) with mouse feeder cells reprograms epithelial cells into stem-like cells with the following properties: 1) indefinite self-renewal, 2) the ability to differentiate into mor mature cells from the tissue of origin when removed from the feeder cells and Y-27632. The cells (called conditionally reprogrammed cells or CRCs) derived from tumors faithfully replicate the original tumor histology in mouse xenografts, while CRCs from normal tissue are nontumorigenic. In a patient with refractory lung tumors (respiratory papillomatosis), CRCs were used to identify vorinostat as a drug that preferentially killed the tumor cells, but not normal tissue. This therapy subsequently led to a dramatic clinical response and reduction in tumor burden for several years. Moreover, the response of the tumor CRCs to drugs in vitro (including both sensitive and resistant agents) recapitulated the patient's responses to therapy in vivo. Based on these exciting results, we hypothesize that CRCs will be an innovative and useful model for PDA to uncover new, effective therapies. We propose to test this hypothesis with the following Specific Aims: 1) To generate CRCs from 20 patient tumors and matched control tissue, and, 2) To use a unique library of >3,000 drugs already approved for use in humans to screen for agents that selectively kill CRCs from PDA, but not normal tissue. This proposal is innovative because: 1) Until now, CRCs have not been generated from PDA samples and will provide a novel tool to study PDA, 2) CRCs have never been used to screen PDA for new therapeutic agents, 3) Because CRCs have properties of stem cells, they may serve as a model for cancer stem cells/tumor-initiator cells and help to identify therapy directed at this unique population of cells, 4) Co-culture with fibroblasts recapitulates the stromal compartment of the tumor microenvironment, 5) Our drug library is diverse, large, and agents are ready for use in patients. At the completion of this grant period, we expect to have a list of new drugs that are cytotoxic to PDA cells to test in preclinical studies and translate into clinical trials led by our clinical collaborators. This strategy could provide a new paradigm for personalized therapy for PDA.