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

• 批准号: 10527325
• 负责人: Munir Tanas
• 金额: $ 34.12万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 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/10527325
• 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; Development; Ependymoma; Epigenetic Process; Gene Fusion; Genetic; Genetic Enhancer Element; Genetic Screening; Genetic Transcription; Genome; Goals; 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; Play; Primary Neoplasm; Proteins; Proteomics; Public Health; Regulation; Research; Role; Signal Transduction Pathway; System; TAZ gene; TFE3 gene; Testing; Transactivation; Transcription Coactivator; Transgenic Organisms; United States National Institutes of Health; Xenograft Model; Xenograft procedure; childhood sarcoma; epigenetic regulation; epigenome; histone acetyltransferase; in vivo; innovation; insight; metaplastic cell transformation; mouse model; mutant; novel; pharmacologic; programs; recruit; sarcoma; small hairpin RNA; targeted treatment; therapeutic target; transcriptome; transcriptome sequencing; tumor growth; tumor initiation; tumorigenesis; validation studies

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.

项目总结/摘要 TAZ/雅普是Hippo通路的转录效应子和原癌蛋白，Hippo通路是一种信号转导通路， 在包括肉瘤在内的多种癌症中调节肿瘤生长和转移的途径。虽然已经发生了很多事情 关于TAZ和雅普的上游调控尚未发现，关于表观遗传调控知之甚少 他们的转录程序，代表了一个显着的知识差距。为了弥补这一差距，我们利用 TAZ-CAMTA 1（TC）和YAP-TFE 3（YT）融合蛋白。TC和YT是上皮样细胞中的驱动癌蛋白 血管内皮瘤（EHE），一种发生于青少年的肉瘤（对PA-16-251，基因融合， 小儿肉瘤）。TC和YT，TAZ和雅普的过度活化形式，是相关的，因为基因融合 是癌症中最常见的TAZ/雅普遗传改变类型。我们提出这项建议的目的是 确定TC和YT是否通过改变表观基因组进一步刺激TAZ/雅普转录。我们的中央 假设TC和YT招募了YEATS 2和ZZZ 3，它们是含Ada 2a（ATAC）组蛋白的一部分 乙酰转移酶复合物，其增强TAZ/雅普致癌转录程序。这一假设 由两种连续的无偏方法支持：BioID质谱法，然后是shRNA筛选， 鉴定YEATS 2和ZZZ 3是TC驱动的锚定非依赖性生长最关键的蛋白质。我们 我计划利用TC（主要的融合蛋白）来检验我们的中心假设，具体目标如下 在EHE中（85%的EHE）： 目的1）确定YEATS 2和ZZZ 3如何改变TAZ-CAMTA 1转录程序。 我们将进行ChIP-seq并确定TC是否与基因组上的YEATS 2和ZZZ 3共定位。RNA-seq 将在表达和不表达YEATS 2或ZZZ 3的TC表达细胞系中进行，以鉴定它们的 对TC转录组的影响。 目的2）鉴定TAZ-CAMTA 1、YEATS 2和ZZZ 3的细胞转化所需结构域。 YEATS 2和ZZZ 3与TC的相互作用将通过利用缺失的免疫共沉淀来剖析 变种人突变体将在我们开发的锚定非依赖性生长试验中进一步研究。 目的3）确定YEATS 2和ZZZ 3与TAZ-CAMTA 1在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YEATS 2/ZZZ 3/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作用。 该建议是创新的，因为它利用TC和YT以及两个系列无偏的方法， 鉴定YEATS 2/ZZZ 3/ATAC为TAZ/雅普转录组的新型表观遗传调节因子，扩展了 重点领域和解决知识的重大差距。该项目意义重大，因为它确定了 YEATS 2和ZZZ 3作为肉瘤中的新型癌蛋白和驱动因子，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，该途径目前缺乏治疗靶点。