The role of TWIST1 in smooth muscle cells during atherosclerosis

TWIST1在动脉粥样硬化过程中平滑肌细胞中的作用

• 批准号: 10576843
• 负责人: Robert Wirka
• 金额: $ 15.46万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10576843
• 关键词: Adopted; Affect; Aorta; ApoE knockout mouse; Apoptosis; Area; Arterial Fatty Streak; Arteries; Atherosclerosis; Automobile Driving; Award; Bayesian Modeling; Bayesian Prediction; Binding; Biological Models; Biology; Blood Vessels; Cell Culture Techniques; Cell Death; Cell Differentiation process; Cell Lineage; Cell physiology; Cells; Cessation of life; Chromatin; Coronary; Coronary Arteriosclerosis; Coronary artery; Data; Data Set; Development; Diabetes Mellitus; Differentiation Antigens; Disease; EP300 gene; Fellowship; Fibroblasts; Gene Expression; Genes; Genetic Transcription; Genetic study; Goals; Human; Human Genetics; Hypertension; Immunofluorescence Immunologic; Immunohistochemistry; In Situ; Injury; Invaded; Knock-out; Knockout Mice; Learning; Lesion; Link; Maps; Medicine; Mentors; Modeling; Molecular; Monitor; Mus; Nature; Paper; Pathway Analysis; Pathway interactions; Phenotype; Positioning Attribute; Process; Proliferating; Regulation; Research; Research Personnel; Risk Factors; Role; Rupture; Sampling; Serum Response Factor; Shapes; Smooth Muscle Myocytes; TWIST1 gene; Time; Training; Transcriptional Regulation; United States; Variant; Vascular Diseases; Vascular Smooth Muscle; Work; aggressive therapy; blood lipid; career development; causal variant; cell dedifferentiation; cell motility; cell type; conditional knockout; design; differential expression; disorder risk; genome-wide; helix-loop-helix protein E12; human data; human disease; in vivo; insight; migration; myocardin; novel; promoter; response; single-cell RNA sequencing; skills; targeted treatment; transcription factor

The purpose of this award is to provide Dr. Robert Wirka protected time, allowing him to obtain the training necessary to become an independent investigator using human genetic findings to define novel mechanisms driving coronary artery disease (CAD). Career development activities are designed to strengthen Dr. Wirka's skills in three key areas necessary for achieving this research paradigm: i) Linking disease-associated variation to causal genes; ii) Determining how causal genes affect cellular and vascular biology; and iii) Defining the molecular pathways and networks affected by causal genes. His mentor, Dr. Quertermous, will guide and monitor his progress towards independence. During atherosclerosis, vascular smooth muscle cells (SMCs) de- differentiate, proliferate and migrate into the lesion in a process known as “phenotypic modulation”. In a recent first-author paper in Nature Medicine, Dr. Wirka generated single-cell gene expression datasets from diseased mouse and human arteries and characterized the process of SMC phenotypic modulation in unprecedented detail. In the current proposal, Dr. Wirka has used these data to identify the transcription factor TWIST1 as a potential novel regulator of SMC phenotypic modulation. In pathway analysis of the mouse and human gene expression data, TWIST1 is strongly predicted to promote SMC phenotypic modulation. Consistent with this predicted role, TWIST1 gene expression increases in SMCs during phenotypic modulation in the mouse and human single-cell data, and preliminary studies show that TWIST1 inhibits expression of SMC differentiation markers. Importantly, TWIST1 is also strongly associated with multiple vascular diseases, including CAD, in human genetic studies. The proposed studies will determine the role of TWIST1 in vascular SMCs during disease, and determine the cellular and molecular mechanisms by which this occurs. In Aim 1, SMC- specific conditional Twist1 knockout mice will be used to determine how Twist1 affects SMC phenotype within the atherosclerotic lesion: i) assessment of single-cell gene expression will be used to determine the effect of SMC-specific Twist1 knockout on the ability of SMCs to undergo phenotypic modulation during disease and to map the molecular pathways affected by Twist1, and ii) in situ studies of the diseased artery wall will examine the effect of SMC-specific Twist1 knockout on classical aspects of lesion phenotype and the contribution of SMCs to the lesion. In Aim 2, cultured human coronary artery SMCs (HCASMCs) will be used to determine the cellular and molecular effects of TWIST1: i) the effect of TWIST1 perturbation on SMC phenotypes relevant to atherosclerosis such as differentiation, proliferation, migration, invasion, and cell death will be determined, and ii) the detailed molecular mechanism by which TWIST1 inhibits expression of the SMC differentiation marker ACTA2 will be explored. These studies will determine the cellular and molecular mechanisms by which a CAD- associated gene and potential novel SMC master regulator influences disease risk. Importantly, these studies will also provide Dr. Wirka with the final skills necessary to achieve scientific independence.

该奖项的目的是提供Robert Wirka受保护的时间，使他能够获得培训 使用人类遗传发现来定义新机制所需的必要 驱动冠状动脉疾病（CAD）。职业发展活动旨在加强Wirka博士 实现这项研究范式所需的三个关键领域的技能：i）与疾病相关的变异联系 致因果基因； ii）确定因果基因如何影响细胞和血管生物学； iii）定义 受因果基因影响的分子途径和网络。他的心理Quermentous博士将指导并 监视他在独立方面的进步。在动脉粥样硬化期间，血管平滑肌细胞（SMC） 在称为“表型调制”的过程中分化，增殖并迁移到病变中。在最近的一个 Wirka博士在自然医学上的第一作者纸，从解散的 小鼠和人动脉，并表征了前所未有的SMC表型调制过程 细节。在当前的建议中，Wirka博士使用这些数据将转录因子Twist1识别为 SMC表型调制的潜在新型调节剂。在小鼠和人类基因的途径分析中 表达数据，Twist1被强烈预测可促进SMC表型调制。与此一致 预测的作用，在小鼠和 人类的单细胞数据，初步研究表明，Twist1抑制SMC分化的表达 标记。重要的是，Twist1也与包括CAD在内的多种血管疾病密切相关 人类遗传研究。拟议的研究将确定Twist1在血管SMC中的作用 疾病，并确定发生这种情况的细胞和分子机制。在AIM 1中，SMC- 特定的条件扭曲1敲除小鼠将用于确定Twist1如何影响SMC表型 动脉粥样硬化病变：i）单细胞基因表达的评估将用于确定 SMC特异性扭曲1敲除SMC在疾病期间经历表型调节能力的能力和 绘制受扭曲1和ii的分子途径，ii）原位研究将检查解散的动脉壁 SMC特异性扭曲1敲除对病变表型的经典方面的影响和 SMC到病变。在AIM 2中，将使用培养的人类冠状动脉SMC（HCASMC）来确定 扭曲的细胞和分子效应1：i）Twist1扰动对与SMC表型相关的影响 将确定动脉粥样硬化，例如分化，增殖，迁移，侵袭和细胞死亡，并 ii）twist1抑制SMC分化标记的表达的详细分子机制 将探索Acta2。这些研究将确定CAD-的细胞和分子机制 相关的基因和潜在的新型SMC主调节器会影响疾病风险。重要的是，这些研究 还将为Wirka博士提供实现科学独立所需的最终技能。