Identify tumor suppressor driver genes of pancreatic ductal adenocarcinoma

摘要胰腺导管腺癌(Pdac)是最致命的常见癌症，没有早期发现。 方法或有效的治疗方法。了解肿瘤的病理进展和分子机制 本病的早期诊断和治疗对寻找新的诊断和治疗方法具有重要意义。不是 有效的药物被开发出来针对PDAC，KRAS， CDKN2A/p16、TP53和Smad4。因此，识别其他pdac驱动基因对开发至关重要。 治疗靶点和研究耐药性。我们已经建立了一个分类和文化的平台 来自人类供体的正常胰腺腺泡和导管细胞，并证明我们能够 用致癌的KRAS和p16、TP53和Smad4的失活来工程这些细胞，以产生 侵袭性PDAC在异种移植小鼠模型中的应用。我们之前的数据显示，仅致癌的KRAS一项就是 不能产生肿瘤，这表明肿瘤发生需要额外的驱动突变(S)。 在这个拟议的项目中，我们将使用我们前所未有的模型以及基因编辑技术来 在PDAC中筛选~200个潜在的抑癌驱动基因，以确定 为PDAC的启动和发展做出贡献。我们将用这些基因工程的人类 研究驱动程序突变如何影响这些细胞的药物敏感性。为了实现我们的目标 研究目标，提出了两个具体的研究目标：目的1.寻找能够抑制肿瘤的基因。 促进腺泡细胞向肿瘤转化；目的2.评估工程化的腺泡细胞和 导管来源的细胞。我们使用基因工程的正常人腺泡细胞的模型代表了 这是概括PDAC在人类细胞中进展的最好模型之一，因此将导致 成功识别PDAC驱动突变是更好地了解肿瘤的关键一步 生物学和靶向治疗的发展，特别是在精确肿瘤学方面。我们的总体目标是 阐明可能导致新的诊断和诊断的分子机制 可提高存活率和改善患者预后的PDAC的治疗方法。

Pancreatic ductal adenocarcinoma (PDAC) is the most deadly common cancer with no early detection methods or effective treatments. Understanding the pathological progression and molecular mechanisms of this disease is crucial to develop new methods for early diagnosis and treatment of PDAC. No effective drugs are developed to target the most frequently mutated four genes in PDAC, KRAS, CDKN2A/p16, TP53 and SMAD4. Thus, identifying other PDAC driver genes is crucial for developing therapeutic targets and studying drug resistance. We have established a platform to sort and culture normal pancreatic acinar and ductal cells from human donor and demonstrated that we are able to engineer these cells with oncogenic KRAS and inactivation of p16, TP53, and SMAD4 to generate invasive PDAC in a xenograft mouse model. Our previous data showed that oncogenic KRAS alone is not able to generate tumors, suggesting that additional driver mutation(s) are required for tumorigenesis. In this proposed project, we will employ our unprecedented model as well as a gene editing technique to screen ~200 potential tumor suppressor driver genes in PDAC to identify the driver mutations that contribute to the initiation and progression of PDAC. We will use these genetically engineered human cells to investigate how the driver mutations could affect drug sensitivity in these cells. To achieve our research goals, two specific aims are proposed: Aim 1. to Identify tumor suppressor genes that can promote acinar cells to tumors; Aim 2. to assess the therapeutic profile of the engineered acinar and ductal derived cells. Our model of using genetically engineered normal human acinar cells represents one of the best models to recapitulate the progression of PDAC in human cells, and thus will lead to successful identification of PDAC driver mutations, a key step towards better understanding tumor biology and development of targeted therapies, especially for precision oncology. Our overall goal is to elucidate the molecular mechanisms which may lead to the development of new diagnostic and treatment methods for PDAC that can increase survival and improve patient outcomes.