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

• 批准号: 10221636
• 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
• 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/10221636
• 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; Dependence; Development; Disease; Drug Targeting; Enhancers; Epigenetic Process; Evaluation; Gene Expression; Gene Expression Alteration; Gene Expression Profiling; 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 profiling; 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 therapeutic intervention; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; programs; ras Proteins; response; screening; small molecule; small molecule inhibitor; subcutaneous; targeted treatment; transcription factor; transcriptome sequencing; translational cancer research; tumor

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.

项目概要/摘要 胰腺导管腺癌（PDAC）是一种毁灭性的疾病，目前是导致胰腺癌的第四大原因。 美国癌症相关死亡。 KRAS 在大多数 PDAC 中发生突变，并且是主要的 这种疾病的致癌驱动因素。不幸的是，尝试开发针对突变 RAS 蛋白的药物 已不成功，并且单药抑制突变 KRAS 下游的效应途径，例如 作为 RAF-MEK-ERK 或 PI3K 途径，迄今为止也被证明是无效的。有一个未满足的关键需求 新的治疗策略。该提案的总体目标是利用功能遗传学方法 确定 KRAS 突变 PDAC 的新漏洞和治疗策略。我们优化了 CRISPR- 人类 PDAC 细胞系中的 Cas9 基因组规模负选择筛选。鉴于抑制 RAS- 丝裂原激活蛋白激酶（MAPK）信号级联将成为组合的重要支柱 PDAC 的治疗方法，我们将基因组规模的 CRISPR-Cas9 筛选与小分子相结合 抑制 MEK1/2 或 ERK1/2 激酶，以确定新的合成致死靶标，这些靶标表现出更大的作用 MAPK 抑制存在依赖性。这些目标包括许多表观遗传调节因子，例如 MEN1/MLL1 和 PRC2 复合体的成员，以及许多转录因子。此外，我们的 使用 PDAC 细胞系破坏后的综合表观遗传学和转录分析进行初步研究 KRAS 信号传导表明 KRAS 介导表观遗传重编程，从而导致不同的细胞状态 具有潜在的可针对的漏洞。该提案建立在这些初步数据的基础上，并有特定的重点 1) MEN1 和 MLL1 作为合成致死靶点的遗传和药理学验证 MAPK 抑制，2) 表观遗传、转录和功能遗传图谱的综合分析 了解 PDAC 中 MAPK 通路抑制所揭示的关键漏洞。功能性 对这些靶标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 (Andrew Aguirre) 博士由威廉·哈恩 (William Hahn) 博士指导，威廉·哈恩博士是一位医学科学家和专家 功能性癌症遗传学，还将受益于由 Matthew 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， 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为期 5 年的培训计划，该计划将利用 DFCI 和哈佛医学院，包括教学课程、科学会议和专业 发展机会将帮助他实现他的科学和职业目标，即发展一个 独立的胰腺癌研究实验室。