Epigenetic modulation of the TAZ-CAMTA1 transcriptional program by the Ada2a-containing histone acetyltransferase complex

含 Ada2a 的组蛋白乙酰转移酶复合物对 TAZ-CAMTA1 转录程序的表观遗传调节

基本信息

批准号：
10304900
负责人：
Munir Tanas
金额：
$ 34.82万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-12-15 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10304900
关键词：
Acetylation Acetyltransferase Address Adolescent Anchorage-Independent Growth Automobile Driving Biological Assay Cell Line ChIP-seq Chimeric Proteins Co-Immunoprecipitations Complex Coupled Data Ependymoma Epigenetic Process Gene Fusion Genetic Genetic Enhancer Element Genetic Screening Genetic Transcription Genome Goals Growth and Development function Hemangioendothelioma Histologic Histone Acetylation Histones Kinetics Knowledge Life Malignant Neoplasms Mass Spectrum Analysis Medical Mission Mutation Nasopharynx Carcinoma Neoplasm Metastasis Non-Small-Cell Lung Carcinoma Oncogenic Oncoproteins Outcome Pathway interactions Pharmacology Play Primary Neoplasm Proteins Proteomics Public Health Regulation Research Role Signal Transduction Pathway System TFE3 gene Testing Transactivation Transcription Coactivator Transgenic Organisms United States National Institutes of Health Xenograft Model Xenograft procedure base childhood sarcoma epigenetic regulation epigenome histone acetyltransferase in vivo innovation insight metaplastic cell transformation mouse model mutant novel programs recruit sarcoma small hairpin RNA targeted treatment therapeutic target transcriptome transcriptome sequencing tumor growth tumor initiation tumorigenesis validation studies

项目摘要

Project Summary/Abstract TAZ/YAP are transcriptional effectors and proto-oncoproteins of the Hippo pathway, a signal transduction pathway regulating tumor growth and metastasis in multiple cancers including sarcomas. While much has been uncovered with regards to the upstream regulation of TAZ and YAP, little is known about epigenetic regulation of their transcriptional programs, representing a significant gap in knowledge. To address this gap, we utilized the TAZ-CAMTA1 (TC) and YAP-TFE3 (YT) fusion proteins. TC and YT are driving oncoproteins in epithelioid hemangioendothelioma (EHE), a sarcoma arising in adolescents (responsive to PA-16-251, Gene Fusions in Pediatric Sarcomas). TC and YT, hyperactivated forms of TAZ and YAP, are relevant because gene fusions are the most common type of genetic alterations of TAZ/YAP in cancer. Our objective in this proposal is to determine if TC and YT further stimulate TAZ/YAP transcription by altering the epigenome. Our central hypothesis is that TC and YT recruit YEATS2 and ZZZ3, part of the Ada2a-containing (ATAC) histone acetyltransferase complex, which potentiate the TAZ/YAP oncogenic transcriptional programs. This hypothesis is supported by two serial unbiased approaches: BioID mass spectrometry followed by an shRNA screen that identified YEATS2 and ZZZ3 as the proteins most critical for TC driven anchorage independent growth. We plan to test our central hypothesis with the following specific aims utilizing TC, the predominant fusion protein in EHE (85% of EHE): Aim 1) Determine how YEATS2 and ZZZ3 alter the TAZ-CAMTA1 transcriptional program. We will perform ChIP-seq and determine if TC co-localizes with YEATS2 and ZZZ3 on the genome. RNA-seq will be performed in TC expressing cell lines with and without expression of YEATS2 or ZZZ3 to identify their effects on the TC transcriptome. Aim 2) Identify domains of TAZ-CAMTA1, YEATS2, and ZZZ3 required for cellular transformation. The interaction of YEATS2 and ZZZ3 with TC will be dissected by co-immunoprecipitation utilizing deletion mutants. The mutants will be further studied in anchorage independent growth assays we have developed. Aim 3) Determine the contributions of the YEATS2 and ZZZ3 interactions with TAZ-CAMTA1 in vivo. We will use both genetic and pharmacological approaches to inhibit YEATS2/ZZZ3/ATAC function in novel TC transgenic and xenograft sarcoma mouse models and evaluate their effect on tumorigenesis and metastasis. The proposal is innovative because it utilizes TC and YT as well as two serial unbiased approaches to identify YEATS2/ZZZ3/ATAC as novel epigenetic regulators of the TAZ/YAP transcriptome, expanding the focus of the field and addressing a significant gap in knowledge. The project is significant because it identifies YEATS2 and ZZZ3 as novel oncoproteins and drivers in sarcoma and provides a rationale for inhibiting histone modifying complexes as a novel way of targeting the Hippo pathway, which currently lacks a therapeutic target.

项目概要/摘要 TAZ/YAP 是信号转导 Hippo 通路的转录效应子和原癌蛋白调节多种癌症（包括肉瘤）中肿瘤生长和转移的途径。虽然已经有很多 TAZ 和 YAP 的上游调控尚未被发现，但表观遗传调控却知之甚少他们的转录程序，代表了知识上的巨大差距。为了解决这个差距，我们利用 TAZ-CAMTA1 (TC) 和 YAP-TFE3 (YT) 融合蛋白。 TC 和 YT 驱动上皮样细胞中的癌蛋白血管内皮瘤 (EHE)，一种发生于青少年的肉瘤（响应 PA-16-251，基因融合儿童肉瘤）。 TC 和 YT（TAZ 和 YAP 的过度激活形式）是相关的，因为基因融合是癌症中最常见的 TAZ/YAP 基因改变类型。我们在此提案中的目标是确定 TC 和 YT 是否通过改变表观基因组进一步刺激 TAZ/YAP 转录。我们的中央假设 TC 和 YT 招募 YEATS2 和 ZZZ3，它们是含有 Ada2a (ATAC) 组蛋白的一部分乙酰转移酶复合物，增强 TAZ/YAP 致癌转录程序。这个假设得到两种连续无偏方法的支持：BioID 质谱分析，然后是 shRNA 筛选确定 YEATS2 和 ZZZ3 是对 TC 驱动的锚定独立生长最关键的蛋白质。我们计划利用主要融合蛋白 TC 来测试我们的中心假设，具体目标如下在 EHE 中（EHE 的 85%）：目标 1) 确定 YEATS2 和 ZZZ3 如何改变 TAZ-CAMTA1 转录程序。我们将进行 ChIP-seq 并确定 TC 是否与 YEATS2 和 ZZZ3 在基因组上共定位。 RNA测序将在表达和不表达 YEATS2 或 ZZZ3 的 TC 表达细胞系中进行，以鉴定其对 TC 转录组的影响。目标 2) 识别细胞转化所需的 TAZ-CAMTA1、YEATS2 和 ZZZ3 结构域。 YEATS2 和 ZZZ3 与 TC 的相互作用将通过利用缺失的免疫共沉淀进行剖析突变体。这些突变体将在我们开发的不依赖于贴壁的生长测定中进行进一步研究。目标 3) 确定 YEATS2 和 ZZZ3 与 TAZ-CAMTA1 体内相互作用的贡献。我们将使用遗传和药理学方法来抑制新型TC中的YEATS2/ZZZ3/ATAC功能转基因和异种移植肉瘤小鼠模型并评估它们对肿瘤发生和转移的影响。该提案具有创新性，因为它利用了 TC 和 YT 以及两种串行无偏方法来确定 YEATS2/ZZZ3/ATAC 作为 TAZ/YAP 转录组的新型表观遗传调节因子，扩展了领域的重点并解决知识上的重大差距。该项目意义重大，因为它确定了 YEATS2 和 ZZZ3 作为肉瘤中的新型癌蛋白和驱动因素，并为抑制组蛋白提供了理论基础修饰复合物作为靶向 Hippo 途径的新方法，目前该途径缺乏治疗靶点。