Pathological reprogramming of the m6A epitranscriptome in uterine fibroids

子宫肌瘤中 m6A 表观转录组的病理重编程

基本信息

批准号：
10300115
负责人：
Ayman Al-Hendy
金额：
$ 66.13万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-09 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10300115
关键词：
Acute Address Benign Bromodomain Catalytic Domain Cell Death Cell Survival Cells ChIP-seq Chemicals Chromatin Chromosomes Chronic Complex Data Set Development Disease Progression Enhancers Epigenetic Process Etiology Expression Profiling Fibroid Tumor Fibrous capsule of kidney Gene Expression Genetic Determinism Genetic Transcription Genetic Translation Growth Habitual Abortion Health Care Costs Homeostasis In Vitro Infertility Knowledge Leiomyoma Link Measures Mediating Medical Menorrhagia Messenger RNA Methylation Methyltransferase Modification Molecular Morbidity - disease rate Mus Myometrial Onset of illness Operative Surgical Procedures Pathogenesis Pathologic Pathway interactions Patients Pelvic Pain Phenotype Premature Labor RNA RNA Binding RNA immunoprecipitation sequencing RNA methylation Role Safety Testing Therapeutic Therapeutic Intervention Time Toxic effect Transcriptional Regulation Translations Treatment Efficacy Tumor Biology Tumor Tissue Up-Regulation Uterine Fibroids Women&apos s Health Xenograft procedure base cell growth cell transformation comparative effective therapy epigenome epitranscriptome epitranscriptomics histone methylation histone modification human disease human model in vivo inhibitor/antagonist insight mRNA Decay mRNA Stability methylome methylomics mouse model mutant myometrium neoplastic new therapeutic target novel overexpression pre-clinical reproductive organ stem cells transcriptome sequencing tumor tumorigenic

项目摘要

PROJECT SUMMARY/ABSTRACT Uterine fibroids (UFs) are the most important benign neoplastic threat to women’s health worldwide. As no long- term non-invasive treatment option currently exists for UFs, deeper insight into tumor etiology is key to develop more effective therapies. In this regard, while the epi/genetic determinants of UFs have been characterized extensively, their underlying pathogenesis nonetheless remains obscure, implicating additional factors in disease onset. Herein, we propose a novel basis to explain UF development through pathological reprogramming of the myometrial epitranscriptome and offer proof of concept for therapeutic intervention using inhibitors of tumorigenic enhancer activity driven by unanticipated m6A-chromatin crosstalk. As the most abundant internal chemical modification in mRNA, N6-methyladenosine (m6A) is a key determinant of posttranscriptional mRNA fate and thus cell identity and function. Accordingly, disruption of m6A homeostasis is implicated in a diverse range of chronic and acute human disease conditions. However, nothing is known about the role of m6A in the pathogenesis of UFs. We now show that m6A writers METTL3 and RBM15 are aberrantly upregulated in UFs compared to adjacent myometrium (MM). Further, we show that METTL3 depletion triggers UF cell death and global upregulation of transcriptionally repressive histone methylation, linking m6A for the first time with UF tumor biology and revealing its novel crosstalk with the UF epigenome. Integrated RNA methylation and expression profiling in METTL3-deficient UF cells revealed a profoundly altered m6A modification landscape and identified high-confidence m6A-modified mRNA effectors of METTL3-driven UF cell growth and survival. Based on these findings, we hypothesize that aberrant METTL3-dependent RNA methylation reprograms the MM epitranscriptome, leading to epigenetic dysregulation and altered expression of genes that drive UF initiation and progression. Accordingly, we propose that epigenetic inhibitors, through suppression of m6A-driven pro- tumorigenic pathways, will provide therapeutic benefit in UFs. To test these hypotheses we will: (1) Establish the basis of epitranscriptomic reprogramming in UFs. We will comparatively profile the m6A modification landscape of mRNAs and chromosome-associated regulatory RNAs (carRNAs) from paired MM and UF tumor tissues and investigate functional cooperativity between METTL3 and RBM15 in methylomic reprogramming; (2) Delineate the role of METTL3-dependent RNA methylation in fibrotic transformation. We will ask if METTL3, in a manner dependent upon its overexpression and methyltransferase activity, can trigger MM stem cell transformation in vitro and UF tumor formation in vivo; (3) Elucidate the impact of METTL3-dependent RNA methylation on gene expression in UFs. We will assess the global impact of m6A on mRNA stability and translation as well as carRNA- dependent control of chromatin state and transcription; (4) Examine the therapeutic potential of BRD inhibitors in a preclinical mouse model of human UFs. We will evaluate select BRD inhibitors for therapeutic efficacy, safety, and mechanism of anti-tumor activity, including impact on chromatin status and transcription.

项目摘要/摘要子宫纤维（UFS）是对全球妇女健康的最重要的良性肿瘤威胁。没有长期术语非侵入性治疗选项目前存在于UFS，更深入地了解肿瘤病因是发展的关键更有效的疗法。在这方面，虽然已经表征了UF的Epi/遗传决定剂广泛地，它们的潜在发病机理仍然晦涩难懂，暗示了疾病的其他因素发作。在此，我们提出了一个新的基础，以通过病理重编程来解释UF的发展肌层表面参考组，并为使用肿瘤抑制剂提供治疗干预的概念验证由意外的M6A-染色质串扰驱动的增强活性。作为最丰富的内部化学化学 MRNA中的修饰，N6-甲基趋化（M6A）是转录后mRNA命运和因此细胞的身份和功能。彼此之间，在潜水员范围内实施了M6A稳态的破坏慢性和急性人类疾病状况。但是，关于M6A在 UFS的发病机理。我们现在证明M6A作者Mettl3和RBM15在UFS中异常更新与邻近的子宫肌层（MM）相比。此外，我们表明METTL3部署触发了UF细胞死亡和转录反射性组蛋白甲基化的全球上调，首次将M6A与UF肿瘤联系起来生物学并揭示其与UF表观基因组的新颖串扰。综合RNA甲基化和表达 Mettl3缺陷UF细胞中的分析显示，M6A的修饰景观发生了深刻变化并确定高信心M6a修饰的mETTL3驱动的UF细胞生长和存活率的mRNA效应。基于这些调查结果，我们假设异常的METTL3依赖性RNA甲基化重编程MM 同意组，导致表观遗传失调并改变了驱动UF启动和的基因表达进展。根据，我们建议通过抑制M6A驱动的促进剂的表观遗传抑制剂肿瘤途径将为UFS提供热益处。为了检验这些假设，我们将：（1）确定 UFS中的表面转录组重编程的基础。我们将相对介绍M6A修改景观来自成对的MM和UF肿瘤组织的mRNA和与染色体相关的调节RNA（CARRNA）以及研究甲基组重编程中METTL3和RBM15之间的功能协调；（2）描绘 Mettl3依赖性RNA甲基化在纤维化转化中的作用。我们会问Mettl3是否以某种方式取决于其过表达和甲基转移酶活性，可以触发MM干细胞转化体内体外和UF肿瘤形成；（3）阐明Mettl3依赖性RNA甲基化对基因的影响 UFS中的表达。我们将评估M6A对mRNA稳定性和翻译的全球影响以及Carrna- 染色质状态和转录的依赖控制；（4）检查BRD抑制剂的治疗潜力在人类UF的临床前小鼠模型中。我们将评估某些BRD抑制剂的治疗效率，抗肿瘤活性的安全性和机制，包括对染色质状态和转录的影响。