Regulation and Function of Myocardin in Vascular Pathobiology

心肌素在血管病理学中的调控和功能

• 批准号: 9042030
• 负责人: Joseph M Miano
• 金额: $ 38.38万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9042030
• 关键词: 3' Untranslated Regions; Acute; Adopted; Alzheimer' s Disease; Arterial Disorder; Asthma; Atherosclerosis; Bacterial Artificial Chromosomes; Binding; Binding Sites; Biological Assay; Biological Preservation; Biology; Blood Vessels; Boxing; Cell Differentiation process; Cells; Chronic; Competence; Consensus Sequence; Data; Data Set; Development; Disease; Elements; Foam Cells; Foundations; Gene Expression; Gene Targeting; Genes; Genomics; Grant; Growth; Health; Human; Hypertension; Indium; Inflammatory; Injury; Ion Channel; Left; Lipids; Maintenance; Methods; MicroRNAs; Modeling; Molecular; Mus; Mutate; Osteoblasts; Paper; Pathway interactions; Phenotype; Point Mutation; Post-Transcriptional Regulation; Process; Regulation; Regulatory Element; Reporting; Repression; Role; Series; Serum Response Factor; Site; Smooth Muscle; Smooth Muscle Myocytes; Staining method; Stains; Stress; Tamoxifen; Testing; Time; Transcription Repressor/Corepressor; Transgenic Mice; Transplantation; Vascular Diseases; Vascular Smooth Muscle; Vascular remodeling; Visceral; atherogenesis; base; cis acting element; cofactor; combat; design; gain of function; in vivo; inflammatory marker; inhibitor/antagonist; insight; interest; loss of function; macrophage; myocardin; myogenesis; next generation sequencing; novel; novel therapeutic intervention; postnatal; programs; promoter; skeletal; skeletal muscle differentiation; transcriptome sequencing; uptake

DESCRIPTION (provided by applicant): Serum response factor (SRF) and Myocardin (MYOCD) constitute a molecular trigger switch for the activation of a battery of vascular smooth muscle cell (VSMC) cyto-contractile and ion channel genes containing SRF- binding CArG boxes. We first showed that levels of MYOCD (but not SRF) correlate with the degree of VSMC differentiation. Since the VSMC differentiation program is not fixed and subject to adaptation in a number of disease processes where levels of MYOCD change, we have been interested in understanding the regulation of MYOCD expression and its function as a homeostatic switch for the preservation of a quiescent, contractile state in VSMC. Accordingly, we have reported in a series of papers that MYOCD (a) is sufficient for conferring VSMC contractile competence through SRF-dependent changes in contractile and ion channel gene expression; (b) completely represses the program of skeletal muscle differentiation; (c) controls a growing number of microRNAs, including the microRNA143/145 gene; and (d) is positively induced by TGF¿1 through a p38MAPK-dependent pathway. Preliminary data further demonstrate cis elements for a transcriptional repressor and new microRNA as well as functional data showing, for the first time, the favorable effects of MYOCD expression on vascular occlusive disease, lipid uptake, and inflammatory marker expression. Collectively, our growing body of work serves as a critical foundation to test the hypothesis that MYOCD is a homeostatic switch for normal VSMC differentiation. This thesis will be tested through a series of inter-related specific aims designe to elucidate transcriptional and post-transcriptional control of MYOCD expression and the role of MYOCD in vascular remodeling during injury-induced neointimal expansion and atherogenesis. In Aim 1, bacterial artificial chromosome transgenic mice will elucidate the function of novel cis-acting elements leading to activation or repression of MYOCD. In Aim 2, novel MYOCD loss- and gain-of-function mice will directly assess the role of this powerful cofactor in experimental vascular disease processes, including effects on VSMC inflammatory, proliferative, and transdifferentiative states. In Aim 3, genomic studies integrating human and mouse CArGome data we have generated (> 84,000 CArG boxes) with RNA-seq of VSMC where MYOCD is expressed in the absence or presence of SRF will be carried out. Such an analysis will reveal a subset of the CArGome that is responsive to MYOCD as well as new SRF-independent MYOCD target genes of import in vascular disease. Thus, the planned studies will yield new insight into the in vivo regulation of MYOCD expression during normal postnatal development and in vascular disease processes as well as novel information related to MYOCD as a likely inhibitor of vascular remodeling associated with physical injury and atherosclerotic disease. The results of these studies will have enormous applications for devising new therapeutic strategies to combat acute and chronic vascular diseases and perhaps other diseases where MYOCD expression/activity may be altered (e.g., asthma, hypertension, Alzheimer's disease).

描述（由申请人提供）：血清反应因子（SRF）和心肌素（MYOCD）构成分子触发开关，用于激活包含SRF结合CArG盒的血管平滑肌细胞（VSMC）细胞收缩基因和离子通道基因组。我们首先证明 MYOCD（但不是 SRF）水平与 VSMC 分化程度相关。由于 VSMC 分化程序不是固定的，并且会在 MYOCD 水平发生变化的许多疾病过程中进行适应，因此我们一直有兴趣了解 MYOCD 表达的调节及其作为维持 VSMC 静止收缩状态的稳态开关的功能。因此，我们在一系列论文中报道，MYOCD (a) 足以通过收缩和离子通道基因表达的 SRF 依赖性变化赋予 VSMC 收缩能力； (b) 完全抑制骨骼肌分化程序； (c) 控制着越来越多的 microRNA，包括 microRNA143/145 基因； (d) TGFβ1 通过 p38MAPK 依赖性途径正向诱导。初步数据进一步证明了转录抑制子和新 microRNA 的顺式元件，以及功能数据首次显示 MYOCD 表达对血管闭塞疾病、脂质摄取和炎症标志物表达的有利影响。总的来说，我们不断开展的工作为检验 MYOCD 是正常 VSMC 分化的稳态开关这一假设奠定了重要基础。本论文将通过一系列相互关联的具体目标进行测试，旨在阐明 MYOCD 表达的转录和转录后控制以及 MYOCD 在损伤诱导的新内膜扩张和动脉粥样硬化过程中血管重塑中的作用。在目标 1 中，细菌人工染色体转基因小鼠将阐明导致 MYOCD 激活或抑制的新型顺式作用元件的功能。在目标 2 中，新型 MYOCD 功能丧失和获得小鼠将直接评估这种强大辅助因子在实验性血管疾病过程中的作用，包括对 VSMC 炎症、增殖和转分化状态的影响。在目标 3 中，将进行基因组研究，将我们生成的人类和小鼠 CArGome 数据（> 84,000 个 CArG 盒）与 VSMC 的 RNA-seq 相结合，其中 MYOCD 在不存在或存在 SRF 的情况下表达。这样的分析将揭示对 MYOCD 敏感的 CArGome 子集以及在血管疾病中导入的新的不依赖于 SRF 的 MYOCD 靶基因。因此，计划中的研究将对正常出生后发育和血管疾病过程中 MYOCD 表达的体内调节产生新的见解，以及与 MYOCD 作为与身体损伤和动脉粥样硬化疾病相关的血管重塑的可能抑制剂相关的新信息。这些研究的结果将具有巨大的应用价值，可用于设计新的治疗策略来对抗急性和慢性血管疾病，以及可能改变 MYOCD 表达/活性的其他疾病（例如哮喘、高血压、阿尔茨海默病）。

项目成果

Myocardin Family Members Drive Formation of Caveolae.

• DOI: 10.1371/journal.pone.0133931
• 发表时间: 2015
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Krawczyk KK;Yao Mattisson I;Ekman M;Oskolkov N;Grantinge R;Kotowska D;Olde B;Hansson O;Albinsson S;Miano JM;Rippe C;Swärd K
• 通讯作者: Swärd K

Myocardin regulates vascular smooth muscle cell inflammatory activation and disease.

• DOI: 10.1161/atvbaha.114.305218
• 发表时间: 2015-04
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Ackers-Johnson M;Talasila A;Sage AP;Long X;Bot I;Morrell NW;Bennett MR;Miano JM;Sinha S
• 通讯作者: Sinha S

CRISPR-Cas9 genome editing of a single regulatory element nearly abolishes target gene expression in mice--brief report.

• DOI: 10.1161/atvbaha.114.305017
• 发表时间: 2015-02
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Han Y;Slivano OJ;Christie CK;Cheng AW;Miano JM
• 通讯作者: Miano JM

Challenges and Opportunities in Linking Long Noncoding RNAs to Cardiovascular, Lung, and Blood Diseases.

将长期非编码RNA与心血管，肺和血液疾病联系起来的挑战和机遇。

• DOI: 10.1161/atvbaha.116.308513
• 发表时间: 2017-01
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Freedman JE;Miano JM;National Heart, Lung, and Blood Institute Workshop Participants*
• 通讯作者: National Heart, Lung, and Blood Institute Workshop Participants*