Functional interrogation of epigenetic vulnerabilities in KRAS-mutant pancreatic cancer

项目概要/摘要 胰腺导管腺癌（PDAC）是一种毁灭性的疾病，目前是胰腺癌的第四大病因。 美国癌症相关死亡人数KRAS在大多数PDAC中发生突变，并且是PDAC的主要基因。 这种疾病的致癌驱动因素。不幸的是，试图开发针对突变RAS蛋白的药物， 已经不成功，并且突变KRAS下游的效应子途径的单一药剂抑制， 如RAF-MEK-ERK或PI3K通路，迄今为止也已证明无效。有一个关键的未满足的需要， 新的治疗策略。该提案的总体目标是利用功能遗传学方法， 在KRAS突变型PDAC中鉴定新的弱点和治疗策略。我们优化了CRISPR- 人PDAC细胞系中的Cas9基因组规模负选择筛选。考虑到RAS的抑制作用- 有丝分裂原活化蛋白激酶（MAPK）信号级联将成为联合的重要支柱 在PDAC的治疗方法中，我们将基因组规模的CRISPR-Cas9筛选与小分子 抑制MEK 1/2或ERK 1/2激酶，以鉴定新的合成致死靶点， 在MAPK抑制剂存在下的依赖性。这些靶点包括许多表观遗传调节因子，如 MEN1/MLL1和PRC2复合物的成员，以及许多转录因子。此外，我们的 使用PDAC细胞系在破坏后的整合表观遗传和转录谱的初步研究 KRAS信号转导表明，KRAS介导表观遗传重编程，导致不同的细胞状态 有潜在的攻击目标本提案以这些初步数据为基础， 1）MEN1和MLL1作为合成致死靶标的遗传和药理学验证， MAPK抑制，2）表观遗传、转录和功能遗传谱的综合分析， 了解PDAC中MAPK通路抑制反应中揭示的关键漏洞。功能 对这些靶点的验证和机制理解可能会导致新的联合治疗策略， PDAC患者安德鲁·阿吉雷博士是由威廉·哈恩博士，一个物理学家，科学家和专家， 功能性癌症遗传学，并将受益于由马修博士组成的咨询委员会 Meyerson，Ramesh Shivdasani博士和Brian Wolpin博士将共同提供指导， 在癌症生物学、表观遗传学和胰腺癌转化研究方面的合作和专业知识。博士 阿吉雷还制定了一项5年培训计划，将利用 DFCI和哈佛医学院，包括教学课程，科学会议和专业 发展机会，这将有助于他实现他的科学和职业发展的目标， 独立的胰腺癌研究实验室

Project Summary / Abstract Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease and currently the fourth-leading cause of cancer-related death in the United States. KRAS is mutated in the majority of PDAC and is the major oncogenic driver of this disease. Unfortunately, attempts to develop drugs that target mutant RAS proteins have been unsuccessful, and single agent inhibition of effector pathways downstream of mutant KRAS, such as the RAF-MEK-ERK or PI3K pathways, has also proven ineffective to date. There is a critical unmet need for novel therapeutic strategies. The overarching goal of this proposal is to utilize functional genetic approaches to identify novel vulnerabilities and therapeutic strategies in KRAS-mutant PDAC. We have optimized CRISPR- Cas9 genome-scale negative-selection screening in human PDAC cell lines. Given that inhibition of the RAS- mitogen-activated-protein-kinase (MAPK) signaling cascade will be an important backbone for combination therapy approaches in PDAC, we have combined genome-scale CRISPR-Cas9 screening with small molecule inhibition the MEK1/2 or ERK1/2 kinases to identify novel synthetic lethal targets that demonstrate greater dependency in the presence of MAPK inhibition. These targets include a many epigenetic regulators, such as members of the MEN1/MLL1 and PRC2 complexes, as well as numerous transcription factors. Additionally, our preliminary studies using integrative epigenetic and transcriptional profiling of PDAC cell lines upon disruption of KRAS signaling suggest that KRAS mediates epigenetic reprogramming that leads to a distinct cell state with potentially targetable vulnerabilities. This proposal builds on these preliminary data with a specific focus on: 1) genetic and pharmacologic validation of MEN1 and MLL1 as synthetic lethal targets in combination with MAPK-inhibition, 2) integrative analysis of epigenetic, transcriptional and functional genetic profiling to understand the key vulnerabilities unveiled in response to MAPK pathway inhibition in PDAC. Functional validation and mechanistic understanding of these targets may lead to novel combination therapy strategies in PDAC patients. Dr. Andrew Aguirre is mentored by Dr. William Hahn, a physician-scientist and expert in functional cancer genetics, and will also benefit from an advisory committee comprised of Dr. Matthew Meyerson, Dr. Ramesh Shivdasani and Dr. Brian Wolpin, who will collectively provide mentorship, collaboration and expertise in cancer biology, epigenetics and pancreatic cancer translational research. Dr. Aguirre has also formulated a 5-year training plan that will leverage the outstanding resources available at DFCI and Harvard Medical School, including didactic coursework, scientific meetings and professional development opportunities that will assist him in achieving his scientific and career goals of developing an independent pancreatic cancer research laboratory.