KLF4-Dependent Regulation of SMC Differentiation and Phenotypic Switching

SMC 分化和表型转换的 KLF4 依赖性调节

• 批准号: 7768056
• 负责人: Gary K Owens
• 金额: $ 66.42万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-12 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7768056
• 关键词: Acetylation; Actins; Adenoviruses; Angioplasty; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Antigens; Aorta; ApoE knockout mouse; Apolipoprotein E; Apoptotic; Applications Grants; Area; Arterial Fatty Streak; Asthma; Atherosclerosis; Automobile Driving; Binding; Binding Sites; Biological Assay; Blood Vessels; Bone Marrow; Bone Marrow Transplantation; CCL2 gene; Candidate Disease Gene; Cardiovascular Diseases; Carotid Arteries; Catheters; Cell Culture System; Cell Differentiation process; Cell Line; Cells; Collaborations; Complex; Confocal Microscopy; Coronary; Coronary artery; Cytomegalovirus; DNA Binding; Dermal; Development; Diet; Differentiation Antigens; Differentiation and Growth; Disease; Down-Regulation; Electrophoretic Mobility Shift Assay; Elements; Embryo; Endothelial Cells; Epigenetic Process; Exhibits; Fibroblasts; Figs - dietary; Freezing; Frequencies; Gel; Gene Activation; Gene Expression; Gene Targeting; Genes; Globin; Goals; Histone Code; Human; Hypertension; Immunofluorescence Immunologic; In Situ Nick-End Labeling; In Vitro; Indium; Inflammatory; Injury; Intramural Research Program; Knock-out; Knockout Mice; Label; Laboratories; LacZ Genes; Lead; Lesion; Letters; Ligation; Liquid substance; MMP3 gene; Maintenance; Measures; Mediating; Mediator of activation protein; Methylation; Microscopic; Modification; Molecular; Mus; Mutate; Mutation; Neoplasm Metastasis; Nitrogen; Oligonucleotides; Oryctolagus cuniculus; Pathogenesis; Pattern; Peptides; Pericytes; Phospholipids; Platelet Factor 4; Platelet-Derived Growth Factor; Play; Process; Proliferating; Promoter Regions; Property; Rattus; Recombinants; Regulation; Regulatory Element; Relative (related person); Repression; Role; Sampling; Sequence Analysis; Series; Shoulder; Signal Transduction; Small Interfering RNA; Smooth Muscle Myocytes; Somatic Cell; Specificity; Staining method; Stains; Stem cells; System; T-Lymphocyte; Tamoxifen; Testing; Time; Transgenes; Transgenic Mice; Transgenic Organisms; Trees; Validation; Vascular Cell Adhesion Molecule-1; Vascular Diseases; aortic arch; atherogenesis; base; cell type; chromatin immunoprecipitation; cohort; combinatorial; effective therapy; embryonic stem cell; extracellular; fascinate; feeding; femoral artery; gene repression; genome-wide; histone modification; human disease; in vivo; insight; interest; loss of function; macrophage; man; morphometry; mutant; myocardin; neointima formation; novel; pluripotency; promoter; public health relevance; reconstitution; response; self-renewal; stem; tumor

DESCRIPTION (provided by applicant): Alterations in the differentiated state of the smooth muscle cell (SMC) play a key role in the development and progression of a variety of cardiovascular diseases. The long-term goal of this project is to elucidate cellular and molecular mechanisms that control the growth and differentiation of SMC during vascular development, and how these control processes are altered during phenotypic switching of SMC in association with vascular injury and disease. Results of our recent studies have implicated the fascinating pluripotency gene Krupple-Like Factor 4 (KLF4) as a key mediator of SMC phenotypic switching in cultured SMC, and in vivo following carotid ligation injury in conditional KLF4 knockout (KO) mice generated in our lab. Studies in this proposal will test the hypothesis that KLF4 plays a critical role in regulating transitions in the differentiated state of vascular SMC during development, as well as during phenotypic switching following vascular injury or development of atherosclerosis. Aim 1 will determine mechanisms by which KLF4 represses expression of SMC marker genes in vivo with vascular injury including determining if KLF4 mediates repression SMC genes by: a) binding to conserved G/C repressor and/or TCE elements found in most SMC promoters; b) inducing epigenetic modifications associated with transcriptional silencing; c) suppressing expression of the potent SMC-selective SRF co-activator myocardin; and/or d) inducing pluripotency genes. Aim 2 will test the hypothesis that delayed repression of SMC marker genes and exacerbated lesion formation following vascular injury observed in our previous studies in conditional KLF4 knockout mice, were mediated, at least in part, by loss of KLF4 in SMC rather than its loss in EC or macrophages. Studies will include bone marrow transfer, and SMC specific conditional KLF4 KO studies in mice injury. Aim 3 is to determine if conditional or SMC specific conditional knockout of KLF4 alters atherosclerotic intimal lesion size or cellular composition in ApoE knockout mice. Aim 4 is to determine the role of KLF4 in activation of genes such as MMP3, MCP-1, iNOS, and VCAM-1 which are likely to play an important functional role in intimal lesion development and/or progression in response to vascular injury or experimental atherosclerosis. PUBLIC HEALTH RELEVANCE: Abnormal control of the differentiated state (i.e. cell specific properties) of vascular smooth muscle cells (SMC) is known to play a critical role in a number of major diseases including atherosclerosis, asthma, hypertension, and tumor metastasis. Studies in this proposal will provide novel insights into mechanisms that control SMC differentiation in development and disease and may lead to new and more effective therapies.

描述（由申请人提供）：平滑肌细胞（SMC）差异化状态的变化在多种心血管疾病的发展和发展中起关键作用。该项目的长期目标是阐明控制血管发育过程中SMC生长和分化的细胞和分子机制，以及在SMC与血管损伤和疾病相关的表型转换过程中如何改变这些控制过程。我们最近的研究的结果暗示了引人入胜的多能基因基因Krupple样因子4（KLF4）是培养的SMC中SMC表型转换的关键介体，在我们的实验室中产生的有条件KLF4敲除（KO）小鼠中的carotid连接损伤后的体内在体内。该提案中的研究将检验以下假设：KLF4在发育过程中血管SMC的分化状态以及血管损伤或动脉粥样硬化发展后的表型转换过程中起着至关重要的作用。 AIM 1将确定KLF4在体内用血管损伤抑制SMC标记基因表达的机制，包括确定KLF4是否介导了抑制SMC基因： b）诱导与转录沉默相关的表观遗传修饰； c）抑制有效的SMC选择性SRF共激活因子心肌素的表达；和/或d）诱导多能基因。 AIM 2将检验以下假设：在我们先前在条件KLF4敲除小鼠方面的研究中观察到的血管损伤后，SMC标记基因的延迟抑制和加剧的病变形成，至少部分是由于SMC中的KLF4而不是部分介导EC或巨噬细胞。研究将包括骨髓转移和SMC特定的小鼠损伤中的有条件KLF4 KO研究。 AIM 3是确定KLF4的条件或SMC特定条件敲除是否改变了APOE敲除小鼠的动脉粥样硬化内膜病变的大小或细胞组成。 AIM 4是确定KLF4在MMP3，MCP-1，INOS和VCAM-1等基因激活中的作用，这些基因可能在响应血管损伤或响应血管损伤或/或进展中起重要的功能作用实验性动脉粥样硬化。 公共卫生相关性：已知血管平滑肌细胞（SMC）的分化状态（即细胞特异性）的异常控制在包括动脉粥样硬化，哮喘，高血压和肿瘤转移在内的多种主要疾病中起关键作用。该提案中的研究将提供对控制SMC分化和疾病分化的机制的新见解，并可能导致新的，更有效的疗法。