Project 1: Role of the H3K27 demethylase KDM6A in bladder cancer pathogenesis

项目1：H3K27去甲基化酶KDM6A在膀胱癌发病机制中的作用

• 批准号: 10475013
• 负责人: David B Solit
• 金额: $ 45.26万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-11 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10475013
• 关键词: ARID1A gene; Agar; Animal Model; Automobile Driving; Bioinformatics; Biologic Characteristic; Biological; Biological Models; Biology; Bladder; Bladder Neoplasm; Bladder Urothelium; CRISPR/Cas technology; Cancer cell line; Carcinogen exposure; Carcinogens; Cell Line; Cell model; Cells; Chromatin; Clinical; Clinical Trials; Clonality; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Critical Pathways; Data; Development; Disease; Down-Regulation; Epigenetic Process; Event; Evolution; Exposure to; Feedback; Frequencies; Future; Gene Expression Profiling; Generations; Genes; Genetically Engineered Mouse; Genomics; Goals; Heterogeneity; Histologic; Histology; Human; Knock-out; Lesion; MAP Kinase Gene; MAPK Signaling Pathway Pathway; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Mediating; Metastatic to; Mitogen-Activated Protein Kinases; Modeling; Molecular; Mus; Muscle; Mutate; Mutation; Nitrosamines; Non-Invasive Lesion; Oncogene Deregulation; Oncogenes; Organoids; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Play; Property; RAS inhibition; Regulation; Research Personnel; Role; Sampling; Signal Transduction; Therapeutic; Transitional Cell Carcinoma; Tumor Cell Line; Urothelium; Work; base; cancer cell; cancer initiation; cancer type; chemical carcinogen; chromatin remodeling; cohort; epigenetic regulation; exome sequencing; experimental study; genomic profiles; human disease; human tissue; in vivo; in vivo Model; individual patient; inhibitor; insight; loss of function; mouse development; mouse model; mutant; next generation sequencing; novel; novel therapeutic intervention; precision medicine; programs; response; senescence; success; translational goal; tumor; tumor initiation; tumor progression

Project Summary/Abstract: Almost all bladder cancers harbor mutations in genes involved in the regulation of chromatin state. KDM6A is among the most commonly mutated of these chromatin-modifying genes in urothelial cancers, and KDM6A alterations are by far more common in urothelial cancer than in any other human cancer type. Here, we seek to further elucidate the biologic role of KDM6A inactivation in bladder cancer pathogenesis as a prelude to the development of rational therapeutic strategies for patients with KDM6A mutant urothelial cancers. On the basis of preliminary data derived from studies of KDM6A isogenic urothelial cancer cells generated by CRISPR/Cas9-targeted inactivation, we hypothesize that KDM6A inactivation is an early event in bladder cancer pathogenesis that promotes tumor formation by deregulating genes that play key roles in modulating RAS/MAP kinase signaling. This proposal will seek to leverage recent advances in next-generation sequencing, clonality analysis, CRISPR gene editing, and patient-derived organoid and genetically-engineered mouse (GEMM) modeling to generate cellular and murine models of KDM6A-deficient bladder cancer that more accurately mirror the genomic profile, histology, and biologic properties of the human disease. These models and a unique cohort of clinically annotated human bladder cancers and the expertise of our co- investigators on this Program Project will then be used to perform in-depth analyses of the biology and functions of KDM6A in bladder cancer pathogenesis. Specifically, we will define the timing at which KDM6A alterations arise in bladder cancer (Aim 1); explore the biologic mechanisms whereby KDM6A loss promotes the transformed phenotype and their implications for drug response (Aim 2); and develop oncogene- and carcinogen-induced mouse models of bladder cancer with Kdm6a loss-of-function to study the mechanisms by which KDM6A inactivation promotes tumor initiation and/or progression in vivo (Aim 3). A long-term translational goal of this work will be to use the insights gained from studies of KDM6A mutant tumors and cell line, organoid and animal models to guide the development of novel therapeutic approaches. Integration: The proposed studies are highly complementary with Project 2, which is focused on the chromatin remodeling gene ARID1A, which is often co-altered in bladder cancers with KDM6A, and with Project 3, which is studying cancer evolution, heterogeneity, and drug response in novel patient-derived organoid models, several of which harbor KDM6A mutations. The success of this project will require close collaboration with all three Cores. Specifically, Core A will provide assistance with the processing and analysis of human tumors for Aim 1 and aid in the histologic and molecular characterization of bladder tumors that arise in GEMMs in Aim 3; Core B will provide KDM6A mutant and wildtype human organoid cell lines for Aim 2 and assist with the generation of GEMMs with targeted deletion of Kdm6a in the urothelium for Aim 3; and Core C will provide critical bioinformatic and statistical support for all Aims, and assist with program integration.

项目摘要/摘要： 几乎所有膀胱癌都含有参与染色质状态调节的基因突变。 KDM6A 是尿路上皮癌中这些染色质修饰基因中最常见的突变之一，并且 迄今为止，KDM6A 改变在尿路上皮癌中比在任何其他人类癌症类型中更为常见。这里， 我们寻求进一步阐明 KDM6A 失活在膀胱癌发病机制中的生物学作用 为 KDM6A 突变尿路上皮患者制定合理治疗策略的序幕 癌症。基于 KDM6A 同基因尿路上皮癌细胞研究的初步数据 由 CRISPR/Cas9 靶向失活产生，我们假设 KDM6A 失活是一个早期事件 在膀胱癌发病机制中，通过解除在膀胱癌中起关键作用的基因的调控来促进肿瘤形成 调节 RAS/MAP 激酶信号传导。该提案将寻求利用下一代技术的最新进展 测序、克隆性分析、CRISPR 基因编辑以及源自患者的类器官和基因工程 小鼠 (GEMM) 建模，生成 KDM6A 缺陷型膀胱癌的细胞和小鼠模型 更准确地反映人类疾病的基因组图谱、组织学和生物学特性。这些 模型和一组独特的临床注释人类膀胱癌以及我们共同的专业知识 然后，该计划项目的研究人员将用于对生物学和 KDM6A 在膀胱癌发病机制中的功能。具体来说，我们将定义 KDM6A 的时序 膀胱癌发生改变（目标 1）；探索 KDM6A 缺失促进的生物学机制 转化的表型及其对药物反应的影响（目标 2）；并发展癌基因和 致癌物诱导的 Kdm6a 功能丧失的小鼠膀胱癌模型，以研究其机制 KDM6A 失活会促进体内肿瘤的发生和/或进展（目标 3）。一个长期的 这项工作的转化目标是利用从 KDM6A 突变肿瘤和细胞研究中获得的见解 线、类器官和动物模型，以指导新型治疗方法的开发。 整合：拟议的研究与项目 2 高度互补，项目 2 的重点是 染色质重塑基因 ARID1A，在膀胱癌中通常与 KDM6A 共同改变，并且 项目 3，正在研究新型患者来源的癌症演变、异质性和药物反应 类器官模型，其中一些含有 KDM6A 突变。该项目的成功需要密切 与所有三个核心的合作。具体来说，核心A将提供处理和分析方面的协助 目标 1 的人类肿瘤的研究，并帮助对出现的膀胱肿瘤进行组织学和分子表征 目标 3 中的 GEMM； Core B将为Aim 2和Aim 2提供KDM6A突变型和野生型人类类器官细胞系 协助生成 GEMM，并针对目标 3 删除尿路上皮中的 Kdm6a；和核心C 将为所有目标提供关键的生物信息学和统计支持，并协助项目整合。