Investigation of therapeutic modulators of apoptotic priming in pancreatic cancer

描述（由申请人提供）：胰腺癌是一种致命的疾病，即使是最具侵略性的细胞毒性化疗方案通常也难以治疗。胰腺癌化疗的最新进展使缓解率有所改善;然而，大多数患者仍然没有从治疗中获益。响应 化疗通常通过细胞凋亡发生，这是一种程序性细胞死亡。我们的目标是鉴定那些功能丧失会增强胰腺癌细胞对 凋亡BH 3谱是癌细胞凋亡敏感性或“引发”的良好表征的上游测量，并且已显示与某些癌症类型中对化疗的反应和疾病预后相关。我们将利用一种称为RNAi-BH 3筛选（RiB筛选）的新筛选方法，该方法将使用混合慢病毒递送的shRNA的功能丧失遗传筛选与BH 3分析技术相结合。在这种方法中，我们将确定基因的敲低结果在几个胰腺细胞系的细胞凋亡增强启动。基于这种功能丧失的遗传筛选，我们将尝试鉴定已知的靶向这些基因的小分子，并确定它们是否改变凋亡启动和化学敏感性。最后，我们建议通过确定其对新鲜患者来源的胰腺癌样品中的BH 3谱和化学敏感性的影响来评估所鉴定的基因或小分子的潜在临床效用。这些基因和小分子的成功鉴定不仅将提高我们对化疗反应的理解，而且还可以作为新的生物标志物或治疗靶点。

DESCRIPTION (provided by applicant): Pancreatic cancer is a deadly disease that is typically refractory to even the most aggressive cytotoxic chemotherapy regimens. Recent advances in chemotherapy for pancreatic cancer have led to some improvement in response rates; however, the majority of patients still do not substantially benefit from therapy. Responses to chemotherapy commonly occur through apoptosis, a form of programmed cell death. Our goal is to identify genes whose loss of function enhances sensitivity of pancreatic cancer cells to apoptosis. BH3 profiling is a well-characterized upstream measurement of the apoptotic sensitivity or "priming" of cancer cells and has been shown to correlate with response to chemotherapy and disease prognosis in certain cancer types. We will utilize a novel screening approach called RNAi-BH3 Screening (RiB Screening) that combines loss-of-function genetic screening using pooled lentivirally delivered shRNAs with the technology of BH3 profiling. In this approach, we will identify genes whose knockdown results in enhanced priming for apoptosis across several pancreatic cell lines. Based on this loss of function genetic screen, we will attempt to identify known small molecules targeting these genes, and determine whether they alter apoptotic priming and chemosensitivity. Finally, we propose to evaluate potential clinical utility of identified genes or small molecules by determining their effect on BH3 Profiles and chemosensitivity in fresh patient-derived pancreatic cancer samples. The successful identification of such genes and small molecules will not only improve our understanding of chemotherapeutic response, but could also serve as novel biomarkers or therapeutic targets.