Targeting aberrant enhancer landscapes in pancreatic cancer

靶向胰腺癌中的异常增强子景观

• 批准号: 10661753
• 负责人: CHRISTOPHER VAKOC
• 金额: $ 40.48万
• 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/10661753
• 关键词: Binding; 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; Experimental Genetics; Future; Gene Expression Profile; Genes; Genetic Enhancer Element; Genetic Transcription; Genome; Genomic approach; Genomics; Goals; HTATIP gene; Human; Intervention; Knock-out; Lead; Location; Maintenance; Malignant neoplasm of pancreas; Maps; Metastatic Adenocarcinoma; Molecular; Mutation; Neoplasm Metastasis; Organoids; Pancreatic Ductal Adenocarcinoma; Patients; Pattern; Phenotype; Polycomb; Primitive foregut structure; Process; Prognosis; Recurrence; Research; Role; Sampling; Site; Source; Squamous Cell; System; Testing; Transplantation; Tumor Promotion; Up-Regulation; cancer cell; effective therapy; epigenetic therapy; experimental study; functional genomics; gain of function; in vivo; innovation; innovative technologies; interest; loss of function; neoplastic; novel; pancreatic cancer cells; pancreatic cancer patients; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; progenitor; programs; restraint; 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.

项目摘要/摘要 该项目的核心目的是制定策略以表观遗传重新编程胰腺癌细胞为 减少转移性传播。这个目标是基于我们最近的胰腺癌的演示 转移伴随着增强子激活的定型模式。我们暗示了先驱 在这种情况下，转录因子FOXA1是增强子重编程和转移性扩散的驱动因素。在 这项建议，我们试图定义增强子 - 纳斯塔西斯的连接。在这样做的过程中，我们寻求提名新的 表观遗传靶标的类别，可能具有独特的能力消除转移性潜力。在第一个目标 在该提案中，我们将采用一种功能基因组学方法来扰动每个FOXA1调节的基因和 增强剂并确定是否可以抑制FOXA1依赖性转移。这种方法将采用 我们最近在以域为中心的CRISPR筛选中创新的优势，并将加深我们的理解 该表观遗传学计划的促分组组成部分。在该提议的第二个目标中，我们将 研究在FOXA1上调之前发生的增强子重编程过程的最早步骤。 这项努力是我们意外的观察到的，即转移特异性增强剂已经存在于 在胰前胰腺肿瘤细胞中可访问的染色质状态。这表明额外的分子 事件发生在FOXA1上调之前，为转移期间增强子重编程奠定了基础。我们 将研究转录因子FOXA2，我们的实验表明，这是一个关键的书签， 打开次转移性癌细胞中转移特异性增强子。此外，我们将确定如何 抑制性多肉液复合物作用于在FOXA1上调之前限制增强子的激活。通过评估 FOXA2/polycomb扰动的后果，我们将提供转移的概念证明 - 相关增强剂可以在胰腺癌进展的早期阶段消失。最终目标 提案将是将我们的增强剂映射研究扩展到胰腺癌的鳞状可能性，即 这是最近定义的与特别惨淡的预后相关的疾病实体。我们将采用一个 新特征的辅助胰腺癌类器官的补充以比较增强剂曲线 与这种疾病的更古典形式相比，鳞状含量。我们将确定这一点的主调节器 鳞状转录程序，并执行遗传实验以确定此类因素在 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 PDA与独特的表观依赖关系有关。拟议的研究共同提供了 用于开发针对独特增强子配置的表观遗传疗法的机械框架 转移性胰腺癌细胞。