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.

这个奖项的目的是提供博士罗伯特Wirka保护的时间，让他获得培训 有必要成为一个独立的研究人员使用人类基因的发现，以确定新的机制 导致冠状动脉疾病（CAD）。职业发展活动旨在加强Wirka博士的 实现这一研究范式所需的三个关键领域的技能：i）将疾病相关变异联系起来 ii）确定因果基因如何影响细胞和血管生物学;以及iii）定义因果基因的功能。 受致病基因影响的分子途径和网络。他的导师Quertermous博士将指导 监督他走向独立的进程在动脉粥样硬化过程中，血管平滑肌细胞（SMC） 分化、增殖并在称为“表型调节”的过程中迁移到病变中。在最近的一 在《自然医学》上发表的第一作者论文中，Wirka博士从疾病中生成了单细胞基因表达数据集， 小鼠和人的动脉，并表征了SMC表型调节的过程， 详细在目前的提案中，Wirka博士利用这些数据将转录因子TWIST 1鉴定为一种转录因子。 SMC表型调节的潜在新调节剂。在小鼠和人类基因的通路分析中， 根据表达数据，强烈预测TWIST 1促进SMC表型调节。符合本 预测的作用，TWIST 1基因表达增加SMC在表型调制在小鼠和 人单细胞数据和初步研究表明，TWIST 1抑制SMC分化的表达， 标记。重要的是，TWIST 1也与多种血管疾病密切相关，包括CAD， 人类基因研究这项研究将确定TWIST 1在血管平滑肌细胞中的作用， 疾病，并确定其发生的细胞和分子机制。在目标1中，SMC- 特异性条件性Twist 1敲除小鼠将用于确定Twist 1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Twist 1基因敲除对SMC在疾病期间进行表型调节和 绘制受Twist 1影响的分子通路，以及ii）对患病动脉壁的原位研究将检查 SMC特异性Twist 1基因敲除对病变表型的经典方面的影响以及 SMC到病变。在目的2中，培养的人冠状动脉SMC（HCASMC）将用于确定在体外培养的人冠状动脉SMC中的表达。 TWIST 1的细胞和分子效应：i）TWIST 1扰动对SMC表型的效应， 将确定动脉粥样硬化的特征，如分化、增殖、迁移、侵袭和细胞死亡， ii）TWIST 1抑制SMC分化标志物表达的详细分子机制 ACTA 2将被探索。这些研究将确定CAD的细胞和分子机制， 相关基因和潜在的新型SMC主调节因子影响疾病风险。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后技能。