Defining the role of histone H3K4 mono-methyltransferase dysfunction in urothelial carcinoma

定义组蛋白 H3K4 单甲基转移酶功能障碍在尿路上皮癌中的作用

• 批准号: 10522552
• 负责人: Yu Chen
• 金额: $ 60.23万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10522552
• 关键词: Ablation; Affect; Allografting; Archives; Benign; Binding; Bladder; Carcinogens; Cell Line; Chromatin; Clinical; Clonal Expansion; Complex; DNA; Data; Down-Regulation; EZH2 gene; Enhancers; Epigenetic Process; Event; Exhibits; Functional disorder; Gene Expression; Genes; Genetic; Genetic Engineering; Genetic Transcription; Genetically Engineered Mouse; Genomic Segment; Histologic; Histology; Histone H3; Histones; Human; KRASG12D; Kidney; Lesion; Location; Lysine; MLL gene; Malignant Neoplasms; Malignant neoplasm of ureter; Malignant neoplasm of urinary bladder; Maps; Mediating; Messenger RNA; Methyltransferase; Mitogen-Activated Protein Kinase Kinases; Modeling; Molecular; Morbidity - disease rate; Morphology; Mus; Muscle; Mutation; Neoplasm Metastasis; Nitrosamines; Oncogene Activation; Oncogenes; Oncogenic; Organoids; Output; Pathway interactions; Patients; Phosphotransferases; Predisposition; Proteins; RNA; Recurrence; Recurrent disease; Regulation; Risk; Role; Running; Sampling; Secondary to; Signal Transduction; Site; Techniques; Time; Transcriptional Regulation; Transitional Cell; Transitional Cell Carcinoma; Tumor Suppression; Tumor-infiltrating immune cells; Up-Regulation; Ureter; Urethra; Urothelial Cell; Urothelium; allotransplant; base; bladder transitional cell carcinoma; cancer invasiveness; combinatorial; fitness; genetic signature; histone demethylase; histone methylation; histone methyltransferase; kinase inhibitor; loss of function mutation; men; migration; mortality; mouse model; preclinical study; programs; promoter; protein complex; response; single-cell RNA sequencing; stemness; transcription factor; transcriptome; tumor; tumorigenesis; Ablation; Affect; Allografting; Archives; Benign; Binding; Bladder; Carcinogens; Cell Line; Chromatin; Clinical; Clonal Expansion; Complex; DNA; Data; Down-Regulation; EZH2 gene; Enhancers; Epigenetic Process; Event; Exhibits; Functional disorder; Gene Expression; Genes; Genetic; Genetic Engineering; Genetic Transcription; Genetically Engineered Mouse; Genomic Segment; Histologic; Histology; Histone H3; Histones; Human; KRASG12D; Kidney; Lesion; Location; Lysine; MLL gene; Malignant Neoplasms; Malignant neoplasm of ureter; Malignant neoplasm of urinary bladder; Maps; Mediating; Messenger RNA; Methyltransferase; Mitogen-Activated Protein Kinase Kinases; Modeling; Molecular; Morbidity - disease rate; Morphology; Mus; Muscle; Mutation; Neoplasm Metastasis; Nitrosamines; Oncogene Activation; Oncogenes; Oncogenic; Organoids; Output; Pathway interactions; Patients; Phosphotransferases; Predisposition; Proteins; RNA; Recurrence; Recurrent disease; Regulation; Risk; Role; Running; Sampling; Secondary to; Signal Transduction; Site; Techniques; Time; Transcriptional Regulation; Transitional Cell; Transitional Cell Carcinoma; Tumor Suppression; Tumor-infiltrating immune cells; Up-Regulation; Ureter; Urethra; Urothelial Cell; Urothelium; allotransplant; base; bladder transitional cell carcinoma; cancer invasiveness; combinatorial; fitness; genetic signature; histone demethylase; histone methylation; histone methyltransferase; kinase inhibitor; loss of function mutation; men; migration; mortality; mouse model; preclinical study; programs; promoter; protein complex; response; single-cell RNA sequencing; stemness; transcription factor; transcriptome; tumor; tumorigenesis

Project Summary/Abstract Urothelial carcinoma (UC) involves the urothelial cells that line the bladder, kidney and ureters and is a major cause of morbidity and mortality in the US, especially in men. Bladder UC can be clinically separated into nonmuscle invasive (NMIBC) and muscle invasive (MIBC). MIBC accounts for the vast majority of metastasis and mortality, having only a ~50% cure rate. Patients with treated NMIBC are at risk of recurrence or progression to MIBC at prior or de novo sites. Over half of urothelial cancer, regardless of site of origin, harbor loss of function mutations in the histone demethylase KDM6A (UTX) and in two highly homologous histone methyltransferases KMT2C (MLL3) and KMT2D (MLL4). These proteins form the MLL3/4-COMPASS (COMplex of Proteins ASsociated with Set1)-like complex that regulate enhancer function, partly through methylation of histones at enhancers. Enhancers are regions of DNA that regulate lineage specific transcriptional programs. Recent studies have shown that patients with two urothelial carcinomas in far away sites (ureter and bladder) harbor the same COMPASS-like mutation. Further sequencing of histologically benign urothelium identify frequent mutations in the complex at expand over time. Our hypothesis is that these mutations under “field-cancerization” of the urothelium. Our lab has generated a genetically engineered mouse model with deletion of Kmt2c, Kmt2d, or the combination in the urothelium. The urothelium of these mice exhibit no histologic abnormalities. However, transcriptome analysis shows the urothelium exhibit increased stemness and functional studies show they exhibit increased organoid forming abilities. When crossed into the Pten conditional deletion mouse, there was robust cooperativity in tumorigenesis. The overall objective of this proposal is to utilize our recently generated mouse models of urothelial this COMPASS-like complex loss to mechanistically understand its role in tumor urothelial suppression. Specifically, in Aim 1, we seek to determine the stemness, clonal dynamics, oncogene and carcinogen susceptibility of urothelium harboring mutations in this COMPASS-like complex, using lineage tracing, organoid culture, and single-cell RNA-sequencing. In Aim 2, we seek to determine the functional interplay between MLL3/4-COMPASS dysfunction and oncogene activation. In Aim 3, we will seek to define how loss of Kmt2c and Kmt2d in urothelial cells affect enhancer and promoter function. Active enhancers are genomic regions of open chromatin with transcription factor binding, divergent transcription of enhancer RNA, and looping to promoters. We will study each step by global mapping of histone marks, chromatin accessibility, mRNA transcription of associated gene and looping to promoters using state-of the art epigenetics techniques.

项目摘要/摘要 尿路上皮癌（UC）涉及在膀胱，肾脏和输尿管上排列的尿路上皮细胞，并且是主要的 美国的发病率和死亡率的原因，尤其是在男性中。膀胱UC可以在临床上分离成 非肌肉入侵（NMIBC）和肌肉入侵（MIBC）。 MIBC占绝大多数转移 和死亡率，只有约50％的治愈率。接受治疗的NMIBC患者有复发的风险或 在先前或从头站点的MIBC进展。港口，不论起源部位如何 组蛋白脱甲基酶KDM6A（UTX）和两个高度同源组蛋白中功能突变的丧失 甲基转移酶KMT2C（MLL3）和KMT2D（MLL4）。这些蛋白质形成MLL3/4-Compass （与set1相关的蛋白质的复合物）类似调节增强子功能的复合物，部分地通过 组蛋白在增强剂处的甲基化。增强子是调节特定谱系的DNA区域 转录程序。最近的研究表明，遥远的两名尿路癌患者 位置（输尿管和膀胱）具有相同的指南针般的突变。组织学的进一步测序 随着时间的推移，良性尿路上皮在综合体中经常识别突变。我们的假设是这些 尿液的“现场癌化”下的突变。我们的实验室已经生成了一条通用的鼠标 删除KMT2C，KMT2D或尿皮上皮中的组合模型。这些小鼠的尿路上皮 没有组织学异常。但是，转录组分析表明尿铺上皮表现出增加 干性和功能研究表明，它们暴露了增强的器官形成能力。当越过 PTEN条件缺失小鼠，在肿瘤发生中有强大的配位。总体目标 建议是利用我们最近生成的尿路上皮的鼠标模型，这种类似罗盘的复杂损失 机械学上了解其在肿瘤尿路上皮抑制中的作用。具体来说，在AIM 1中，我们试图确定 尿液中携带突变的尿液，克隆动力学，癌基因和致癌物的敏感性 这种指南针般的复合物，使用谱系跟踪，器官培养和单细胞RNA测序。目标 2，我们试图确定MLL3/4-兼容功能障碍与癌基因之间的功能相互作用 激活。在AIM 3中，我们将寻求定义尿路上皮细胞中KMT2C和KMT2D的损失如何影响增强子和 启动子功能。活性增强子是带有转录因子结合的开放染色质的基因组区域， 增强子RNA的发散转录，并向启动子循环。我们将通过全球映射研究每个步骤 组蛋白标记，染色质可及性，相关基因的mRNA转录和循环启动子 使用最先进的表观遗传学技术。