Mediator complex in cardiac development and function

心脏发育和功能的介导复合物

基本信息

批准号：
10199010
负责人：
Xi Fang
金额：
$ 24.52万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10199010
关键词：
Address Adult Affect Binding Bioinformatics Birth Cardiac Cardiac Myocytes Cardiac development Cardiology Cardiomyopathies Cardiovascular Diseases Career Mobility ChIP-seq Complex Congenital Heart Defects DNA Binding DNA Polymerase II Development Dilated Cardiomyopathy Elements Enhancers Ensure Event Exhibits Family Gene Expression Genes Genetic Transcription Genetic study Goals Head Heart Heart Abnormalities Heart Diseases Heart failure Homeostasis Human Hypertrophy Individual Knock-out Knockout Mice Lead Light MED25 gene Maintenance Mediator of activation protein Mentors Mentorship Missense Mutation Modeling Molecular Morphogenesis Morphology Mus Mutation Oranges Participant Pathologic Patients Phase Phenotype Phosphotransferases Physiological Physiology Play RNA Polymerase II Regulation Regulator Genes Research Research Project Grants Role Shapes Shprintzen syndrome Signal Transduction Structure Tail Time Training Transcript Transcription Coactivator Transcription Initiation Transcriptional Regulation cardiogenesis career cell type congenital heart disorder genomic locus heart function insight mouse genetics mouse model novel programs promoter skills transcription factor transcriptome sequencing

Address Adult Affect Binding Bioinformatics Birth Cardiac Cardiac Myocytes Cardiac development Cardiology Cardiomyopathies Cardiovascular Diseases Career Mobility ChIP-seq Complex Congenital Heart Defects DNA Binding DNA Polymerase II Development Dilated Cardiomyopathy Elements Enhancers Ensure Event Exhibits Family Gene Expression Genes Genetic Transcription Genetic study Goals Head Heart Heart Abnormalities Heart Diseases Heart failure Homeostasis Human Hypertrophy Individual Knock-out Knockout Mice Lead Light MED25 gene Maintenance Mediator of activation protein Mentors Mentorship Missense Mutation Modeling Molecular Morphogenesis Morphology Mus Mutation Oranges Participant Pathologic Patients Phase Phenotype Phosphotransferases Physiological Physiology Play RNA Polymerase II Regulation Regulator Genes Research Research Project Grants Role Shapes Shprintzen syndrome Signal Transduction Structure Tail Time Training Transcript Transcription Coactivator Transcription Initiation Transcriptional Regulation cardiogenesis career cell type congenital heart disorder genomic locus heart function insight mouse genetics mouse model novel programs promoter skills transcription factor transcriptome sequencing

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项目摘要

Mediator (MED) is a multi-subunit complex that plays a central role in transcription initiation by integrating regulatory signals from gene-specific transcriptional activators to RNA polymerase II. Genetic studies have identified mutations in specific MED subunits to be associated with human cardiac diseases. Specifically, deletions in MED15 are common in patients with DiGeorge/velocardiofacial syndrome, which typically includes congenital heart defects. A missense mutation in MED30 causes cardiomyopathy in mice. Emerging evidence suggests a critical role for “specialized” MED subunits in the regulation of temporal- and/or spatial-specific gene expression. However, little is known as to the subunit composition of MED in cardiomyocytes (CMs) at distinct developmental stages, and no studies have examined enrichment of MED complexes harboring individual “specialized” MED subunits at distinct genomic loci, as well as the specific transcription factors (TFs) they directly interact with to anchor them to functionally critical enhancer/promoter elements during heart development, or during the progression of cardiac disease. We found that transcript levels of MED subunits fluctuate within CMs during development. To gain further understanding into MED subunit specific roles in CMs, I generated novel mouse models with cardiac-specific knockout (cKO) of selected MED subunits. MED15, MED25, and MED30 cKO mice showed distinct phenotypes. MED15 cKO mice died immediately after birth, with cardiac defects observed from E12.5, whereas MED30 cKO mice died at E10-10.5 with dilated hearts. Conversely, MED25 cKO mice survived to adulthood and displayed no cardiac defects, demonstrating that not all MED subunits are indispensable for cardiac function. Accordingly, my hypothesis is that MED15 and MED30 subunits exhibit unique functional activities that shape key events in CM transcriptional regulation, morphogenesis, and heart function. My Specific Aims are: Aim 1. (K99) To elucidate stage-specific and locus- specific roles of MED15 or MED30 subunits in developing CMs by analyzing cardiac phenotypes of MED15 and MED30 cKO mice, identifying unique transcriptional regulatory elements enriched for MED15 or MED30 during early heart development, and identifying TFs that specifically interact with MED15 or MED30; and Aim 2. (R00) To understand the pathophysiological functions of MED15 and MED30 subunits during the progression of adult heart disease, including DCM and pathological hypertrophy, by utilizing inducible MED15 and MED30 CM-specific knockout (icKO) mouse models. I have a strong background and training record in mouse genetics and molecular cardiology. My training in the K99 phase will consist of structured mentorship by my primary mentor and complementary co-mentors/consultants, formal coursework, additional training in experimental skills (RNAseq, ChIP-seq and bioinformatics) and a program of career transition. This career plan and research project will ensure my successful transition to independent research, to fulfill my ultimate career goal of understanding the development, progression, and molecular basis of human cardiovascular diseases.

中介（MED）是一种多生产综合体，通过整合在转录启动中起着核心作用从基因特异性转录激活剂到RNA聚合酶II的调节信号。遗传研究已有在特定的MED亚基中鉴定出与人类心脏疾病有关的突变。具体来说， Med15中的缺失在Digeorge/Velocardiofacial综合征的患者中很常见，通常包括先天性心脏缺陷。 Med30的错义突变会导致小鼠心肌病。新兴证据在调节临时和/或空间特异性的调节中，提出了“专业” Med亚基的关键作用基因表达。但是，鲜为人知的是心肌细胞（CMS）中MED的亚基组成独特的发育阶段，没有研究检查携带的Med综合体的富集在不同的基因组基因座以及特定的转录因子（TFS）处的单个“专业” MED亚基它们直接与锚定为锚定在功能上关键的增强子/启动子元素中发育，或心脏病的进展。我们发现Med亚基的转录级水平开发过程中CMS内波动。进一步了解Med亚基在 CMS，我生成了带有心脏特异性基因敲除（CKO）的新型小鼠模型。 Med15，Med25和Med30 CKO小鼠显示出不同的表型。 Med15 CKO小鼠立即死亡出生，从E12.5观察到心脏缺陷，而Med30 CKO小鼠死于E10-10.5 心。相反，Med25 CKO小鼠幸存到成年，没有表现出心脏缺陷，证明并非所有的MED亚基对于心脏功能都是必不可少的。根据我的假设是Med15 Med30亚基表现出独特的功能活动，这些活动塑造了CM转录调控中的关键事件，形态发生和心脏功能。我的具体目的是：AIM 1。（K99）阐明特定于阶段的阶段和基因座 - 通过分析的Med15的心脏表型，Med15或Med30亚基在开发CMS中的特定作用和Med30 CKO小鼠，确定富含Med15或Med30的独特转录调节元件在早期心脏发育期间，并确定与Med15或Med30特别相互作用的TF；和目标 2。（R00）了解Med15和Med30亚基的病理生理功能通过使用诱导Med15，包括DCM和病理肥大在内的成人心脏病的进展和Med30 cm特异性敲除（ICKO）小鼠模型。我的背景和训练记录很强小鼠遗传学和分子心脏病学。我在K99阶段的培训将包括结构化的心态我的主要导师和完成的联合会/顾问，正式课程，额外的培训实验技能（RNASEQ，CHIP-SEQ和BioInformatics）和职业过渡计划。这个职业计划研究项目将确保我成功过渡到独立研究，以实现我的最终职业了解人类心血管疾病的发展，进展和分子基础的目标。