Functional interrogation of epigenetic vulnerabilities in KRAS-mutant pancreatic cancer

KRAS 突变胰腺癌表观遗传脆弱性的功能研究

基本信息

批准号：
9370987
负责人：
Andrew James Aguirre
金额：
$ 17.71万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9370987
关键词：
Advisory Committees Biological Models Biology CRISPR screen CRISPR/Cas technology Cancer Biology Cancer Etiology Cell Line Cell Proliferation Cell Survival Cells Cessation of life ChIP-seq Clinical Research Clinical Trials Collaborations Combined Modality Therapy Complex Data Dependency Development Disease Drug Targeting Employee Strikes Enhancers Epigenetic Process Evaluation Gene Expression Gene Expression Alteration Gene Targeting Genetic Genetic Screening Genetic Transcription Genetic study Goals Human In Vitro Investigation KRAS2 gene Knock-out Laboratory Research Lead MAP2K1 gene MAPK3 gene MEK inhibition MEKs MLL gene Malignant neoplasm of pancreas Malignant neoplasm of prostate Maps Mediating Menin Mentors Mentorship Mitogen-Activated Protein Kinases Modeling Mutate Oncogenic Pancreatic Ductal Adenocarcinoma Pathway interactions Patients Pharmaceutical Preparations Pharmacology Pharmacology Study Phosphotransferases Physicians Pre-Clinical Model RAS inhibition Recurrence Resources Role Scientist Signal Pathway Signal Transduction Therapeutic Training Transcription Alteration United States Validation Vertebral column Work Xenograft procedure base cancer genetics career design epigenetic regulation epigenomics established cell line genetic approach genetic profiling genome-wide histone methyltransferase in vivo in vivo Model inhibitor/antagonist leukemia medical schools meetings member mutant novel novel therapeutics programs ras Proteins response screening small molecule small molecule inhibitor subcutaneous targeted treatment transcription factor transcriptome sequencing translational cancer research tumor

Advisory Committees Biological Models Biology CRISPR screen CRISPR/Cas technology Cancer Biology Cancer Etiology Cell Line Cell Proliferation Cell Survival Cells Cessation of life ChIP-seq Clinical Research Clinical Trials Collaborations Combined Modality Therapy Complex Data Dependency Development Disease Drug Targeting Employee Strikes Enhancers Epigenetic Process Evaluation Gene Expression Gene Expression Alteration Gene Targeting Genetic Genetic Screening Genetic Transcription Genetic study Goals Human In Vitro Investigation KRAS2 gene Knock-out Laboratory Research Lead MAP2K1 gene MAPK3 gene MEK inhibition MEKs MLL gene Malignant neoplasm of pancreas Malignant neoplasm of prostate Maps Mediating Menin Mentors Mentorship Mitogen-Activated Protein Kinases Modeling Mutate Oncogenic Pancreatic Ductal Adenocarcinoma Pathway interactions Patients Pharmaceutical Preparations Pharmacology Pharmacology Study Phosphotransferases Physicians Pre-Clinical Model RAS inhibition Recurrence Resources Role Scientist Signal Pathway Signal Transduction Therapeutic Training Transcription Alteration United States Validation Vertebral column Work Xenograft procedure base cancer genetics career design epigenetic regulation epigenomics established cell line genetic approach genetic profiling genome-wide histone methyltransferase in vivo in vivo Model inhibitor/antagonist leukemia medical schools meetings member mutant novel novel therapeutics programs ras Proteins response screening small molecule small molecule inhibitor subcutaneous targeted treatment transcription factor transcriptome sequencing translational cancer research tumor

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项目摘要

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.

项目摘要 /摘要胰腺导管腺癌（PDAC）是一种毁灭性疾病，目前是第四个领先的原因美国与癌症有关的死亡。 KRAS在大多数PDAC中被突变，是主要的这种疾病的致癌驱动力。不幸的是，试图开发靶向突变体RAS蛋白的药物一直没有成功，并且单药抑制突变体Kras下游的效应途径，此类作为RAF-Mek-ERK或PI3K途径，迄今也已证明无效。有关键的未满足需求新颖的治疗策略。该提案的总体目标是利用功能遗传方法确定KRAS突变PDAC中的新型脆弱性和治疗策略。我们已经优化了CRISPR- CAS9基因组规模的负筛选在人PDAC细胞系中。考虑到抑制Ras- 有丝分裂原活化蛋白激酶（MAPK）信号级联将是组合的重要骨干 PDAC的治疗方法，我们将基因组尺度CRISPR-CAS9与小分子结合在一起抑制MEK1/2或ERK1/2激酶，以鉴定出新的合成致死靶标，这些靶标显示出更大的在MAPK抑制存在下的依赖性。这些目标包括许多表观遗传调节剂，例如 MEN1/MLL1和PRC2复合物以及许多转录因子的成员。另外，我们的使用综合表观遗传学和转录分析的初步研究，破坏了PDAC细胞系 KRAS信号传导的表明，KRAS介导的表观遗传重编程，导致不同的细胞状态具有潜在的目标漏洞。该提案以这些初步数据为基础，以特定的重点 ON：1）MEN1和MLL1作为合成致死靶标的遗传和药理验证与 MAPK抑制作用，2）表观遗传，转录和功能遗传分析的综合分析了解响应PDAC中MAPK途径抑制的关键漏洞。功能对这些目标的验证和机械理解可能会导致新的组合治疗策略 PDAC患者。安德鲁·阿奎尔（Andrew Aguirre 功能性癌症遗传学，也将从马修博士组成的咨询委员会中受益 Meyerson，Ramesh Shivdasani博士和Brian Wolpin博士，他们将集体提供指导，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 Aguirre还制定了一项为期5年的培训计划，该计划将利用可用的未偿还资源 DFCI和哈佛医学院，包括教学课程，科学会议和专业开发机会将有助于他实现他的科学和职业目标独立的胰腺癌研究实验室。