Targeting aberrant enhancer landscapes in pancreatic cancer

靶向胰腺癌中的异常增强子景观

• 批准号: 9816984
• 负责人: CHRISTOPHER VAKOC
• 金额: $ 41.31万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-02 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9816984
• 关键词: Address; Adenocarcinoma Cell; Biological Assay; CRISPR screen; Cells; Cessation of life; Chromatin; Chromatin Remodeling Factor; Collection; Complement; Complex; Dependence; Development; Diagnosis; Disease; Disseminated Malignant Neoplasm; Ductal Epithelial Cell; Elements; Embryo; Endoderm; Enhancers; Epigenetic Process; Event; Future; Gene Expression Profile; Genes; Genetic; Genetic Enhancer Element; Genetic Transcription; Genome; Genomic approach; Genomics; Goals; HTATIP gene; Human; Intervention; Knock-out; Lead; Location; Maintenance; Malignant neoplasm of pancreas; Metastatic Adenocarcinoma; Modeling; Molecular; Mutation; Neoplasm Metastasis; Organoids; Pancreatic Ductal Adenocarcinoma; Patients; Pattern; Phenotype; Polycomb; Primitive foregut structure; Process; Recurrence; Research; Role; Sampling; Site; Source; Squamous Cell; System; Testing; Transplantation; Up-Regulation; base; cancer cell; effective therapy; epigenetic therapy; experimental study; functional genomics; in vivo; innovation; innovative technologies; interest; loss of function; neoplastic; novel; outcome forecast; pancreatic cancer cells; pancreatic cancer patients; progenitor; programs; targeted treatment; therapeutic target; trait; transcription factor; transcriptome; tumor progression

Project Summary/Abstract The central goal of this project is to develop strategies to epigenetically reprogram pancreatic cancer cells to diminish metastatic spread. This objective is based on our recent demonstration that pancreatic cancer metastasis is accompanied by a stereotypical pattern of enhancer activation. We implicated the pioneer transcription factor FOXA1 as a driver of enhancer reprogramming and of metastatic spread in this context. In this proposal, we seek to define the enhancer-metastasis connection. In doing so, we seek to nominate a new class of epigenetic targets, which might be uniquely capable of eliminating metastatic potential. In the first Aim of this proposal, we will employ a functional genomics approach to perturb every FOXA1-regulated gene and enhancer and determine whether FOXA1-dependent metastasis can be suppressed. This approach will take advantage of our recent innovations in domain-focused CRISPR screening and will deepen our understanding of the pro-metastatic components of this epigenetic program. In the second Aim of this proposal, we will investigate the earliest steps of the enhancer reprogramming process that occur prior to FOXA1 upregulation. This effort builds from our unexpected observation that metastasis-specific enhancers are already present in an accessible chromatin state in pre-metastatic pancreatic tumor cells. This suggests that additional molecular events occur prior to FOXA1 upregulation to set the stage for enhancer reprogramming during metastasis. We will investigate the transcription factor FOXA2, which our experiments suggest is the critical bookmark that opens up metastasis-specific enhancers in pre-metastatic cancer cells. In addition, we will determine how repressive Polycomb complexes act to restrain enhancer activation prior to FOXA1 upregulation. By evaluating the consequences of FOXA2/Polycomb perturbation, we will provide a proof-of-concept that metastasis- associated enhancers can be effaced at early stages of pancreatic cancer progression. The final Aim of this proposal will be to extend our enhancer mapping studies into the squamous-subtype of pancreatic cancer, which is a recently defined disease entity associated with a particularly dismal prognosis. We will employ a newly characterized complement of patient-derived pancreatic cancer organoids to compare enhancer profiles of squamous-subtype versus the more classical form of this disease. We will identify master-regulators of this squamous transcriptional program, and perform genetic experiments to determine the role of such factors in promoting tumor progression and metastatic spread. We will also determine whether squamous cell identity in PDA is associated with unique epigenetic dependencies. Collectively, the proposed research will provide a mechanistic framework for developing epigenetic therapies that target the unique enhancer configuration of metastatic pancreatic cancer cells.

项目概要/摘要 该项目的中心目标是制定策略，对胰腺癌细胞进行表观遗传重编程，使其 减少转移扩散。这一目标是基于我们最近的证明，即胰腺癌 转移伴随着增强子激活的典型模式。我们牵连了先锋 转录因子 FOXA1 作为增强子重编程和转移扩散的驱动因素。在 在这个提案中，我们寻求定义增强子与转移的联系。在此过程中，我们寻求提名新的 一类表观遗传靶点，可能具有独特的消除转移潜力的能力。在第一个目标中 根据该提案，我们将采用功能基因组学方法来干扰每个 FOXA1 调节的基因， 增强子并确定是否可以抑制 FOXA1 依赖性转移。这种方法将采取 利用我们最近在以领域为中心的 CRISPR 筛选方面的创新，并将加深我们的理解 该表观遗传程序的促转移成分。在本提案的第二个目标中，我们将 研究 FOXA1 上调之前发生的增强子重编程过程的最早步骤。 这项工作建立在我们意想不到的观察基础上，即转移特异性增强子已经存在于 转移前胰腺肿瘤细胞中可接近的染色质状态。这表明额外的分子 事件发生在 FOXA1 上调之前，为转移过程中增强子重编程奠定基础。我们 将研究转录因子 FOXA2，我们的实验表明它是关键的书签 在转移前的癌细胞中打开转移特异性增强子。此外，我们将确定如何 抑制性 Polycomb 复合物在 FOXA1 上调之前抑制增强子激活。通过评估 FOXA2/Polycomb 扰动的后果，我们将提供转移的概念验证 相关的增强子可以在胰腺癌进展的早期阶段被消除。本次活动的最终目的 建议将我们的增强子图谱研究扩展到胰腺癌的鳞状亚型， 这是最近定义的一种与特别糟糕的预后相关的疾病实体。我们将聘请一名 新鉴定的患者来源的胰腺癌类器官的补充物，用于比较增强子谱 鳞状亚型与该疾病的更经典形式的比较。我们将确定这个的主监管者 鳞状转录程序，并进行基因实验以确定这些因素在 促进肿瘤进展和转移扩散。我们还将确定鳞状细胞身份是否在 PDA 与独特的表观遗传依赖性相关。总的来说，拟议的研究将提供 开发针对独特增强子配置的表观遗传疗法的机制框架 转移性胰腺癌细胞。