Function and Regulation of MEIS1 in Vascular Disease

MEIS1在血管疾病中的功能和调控

基本信息

批准号：
10723084
负责人：
Amr Salem
金额：
$ 11.35万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10723084
关键词：
Alleles Angioplasty Antibodies Arteries Atherosclerosis Award Binding Binding Sites Biological Assay Blood Vessels Cell Cycle Cell Differentiation process Cells ChIP-seq Chromatin Chromosomes Collaborations Coronary Coronary artery Cre driver Data Detection Disease Down-Regulation EVI1 gene Elements Endarterectomy Enhancers Epitopes Event Foundations Gel Gene Expression Genes Genetic Transcription Homeostasis Human Inflammation Injury Knockout Mice LoxP-flanked allele Luciferases MEIS1 gene Maintenance Mentors Methods Mission Mus Mutate Myelogenous National Heart, Lung, and Blood Institute Nuclear Nuclear Accidents Paper Pathologic Pathology Patients Phase Phenotype Positioning Attribute Proliferating Promoter Regions Proteins Pulmonary Hypertension Rattus Regulation Regulatory Element Reporting Research Research Personnel Resources Response Elements Rodent Role Serum Response Factor Signal Pathway Smooth Muscle Myocytes Stenosis Testing Training Transactivation Transcription Coactivator Transcriptional Regulation Validation Vascular Diseases Vascular Smooth Muscle Work atherogenesis biobank career cell dedifferentiation cell motility cell type chromatin immunoprecipitation cofactor combat data mining epigenetic marker epigenome gain of function genome editing genome-wide analysis in vivo inhibitor innovation insight loss of function migration mouse model multiple omics myocardin novel preservation prevent prime editing promoter protein expression restenosis spatiotemporal therapeutic target transcription factor transcriptome transcriptome sequencing transcriptomic profiling transcriptomics vascular injury vascular smooth muscle cell migration vascular smooth muscle cell proliferation virtual

项目摘要

PROJECT SUMMARY Vascular smooth muscle cells (VSMCs), the major cell type of medium- and large-sized arteries, undergo phenotypic switching in vascular diseases such as post-angioplasty restenosis and atherosclerosis. Understanding the key players that promote VSMC plasticity will inform targeting strategies for downstream signaling pathways and nuclear events effecting pathological changes in gene expression. Myocardin (MYOCD) is a potent transcriptional coactivator of the VSMC differentiated state and loss of expression portends vascular pathology. Despite the hundreds of papers describing MYOCD expression and activity in various contexts, its transcriptional regulation in vivo is virtually unknown. I have discovered a binding site for Myeloid Ecotropic Viral Integration Site 1 (MEIS1), a previously unrecognized transcription factor in VSMCs, in the 5’ promoter region of Myocd, and preliminary studies show MEIS1 binding and transactivation of the Myocd gene. MEIS1 is reduced in human VSMCs of atherosclerotic coronary arteries and transcriptomic profiling from Biobank of Karolinska Endoarterctomy (BiKE) showed significant down-regulation of MEIS1 in plaques versus normal arteries as well as in symptomatic versus asymptomatic patients. To date, there has been only one report of MEIS1 in VSMCs and nothing known beyond descriptive findings in pulmonary hypertension. Previous Meis1 loss-of-function and protein expression studies have been difficult to interpret because of the lack of a true null mouse model and faulty commercial antibodies. Preliminary studies below show an increased proliferation and migration of VSMCs with loss of Meis1 function; gain-of-function studies reveal the maintenance of the contractile state. I have generated a versatile mouse model representing the first true null Meis1 allele and its detection at the protein level. An important component of my studies will be the utilization of a new VSMC-restricted Cre driver I have helped characterize (Itga8-CRERT2) for more refined in vivo loss-of-function studies. Additional studies support multiple enhancers that drive VSMC expression of Meis1. Collectively, the preliminary work support my hypothesis that MEIS1 is a VSMC enriched transcription factor that promotes VSMC differentiation via transactivation of Myocd. I propose to test this hypothesis in three aims over the course of this K99/R00 award. The K99 Aim 1 will elucidate the VSMC phenotype with loss of endogenous MEIS1. R00 Aim 2 will elucidate the transcriptional regulation of Myocd in vivo through an evolutionarily-conserved MEIS1-response element using state-of-the-art genome editing in mice. R00 Aim 3 will elucidate the functional cis-regulome of MEIS1 in vascular disease. Completion of the proposed studies will provide new insight into the contribution of MEIS1 to VSMC differentiation and vascular homeostasis and the mechanistic regulation of Meis1 in vascular diseases. Conceptual and experimental training related to multi-omics related research during the K99 phase will help me pursue advanced MEIS1 studies as an independent investigator during my R00 phase and position me well for my first R01 application.

项目摘要血管平滑肌细胞（VSMC）是中型和大型动脉的主要细胞类型血管疾病（如血管成形术后再狭窄和动脉粥样硬化）等血管疾病中的表型转换。了解促进VSMC可塑性的主要参与者将为下游的目标策略提供信息信号通路和核事件影响基因表达的病理变化。心肌（Myocd）是VSMC分化状态的潜在转录共激活因子，表达损失pertends血管病理。尽管有数百篇论文描述了各种情况下的MyOCD表达和活动，但体内的转录调节实际上是未知的。我发现了一个髓样生态病毒的结合位点集成位点1（MEIS1），VSMC中先前未认可的转录因子，在5'启动子区域 MyOCD和初步研究表明MEIS1结合和反式激活的基因。 Meis1减少了在人类VSMC中内部切除术（自行车）显示斑块与正常动脉中MEIS1的显着下调如症状和无症状患者。迄今为止，VSMC中只有一份MEIS1的报告除了肺动脉高压中的描述性发现以外，没有什么知道的。以前的Meis1功能丧失和蛋白质表达研究很难解释，因为缺乏真正的无效小鼠模型和商业抗体故障。下面的初步研究表明，VSMC的增殖和迁移增加 MEIS1功能的损失；功能收益研究揭示了收缩状态的维护。我有生成了代表第一个真实NULL MEIS1等位基因的多功能小鼠模型及其在蛋白质上的检测等级。我的研究的一个重要组成部分是利用新的VSMC限制CRE驱动程序有助于表征（ITGA8-CRERT2），以进行更精致的功能丧失研究。其他研究支持驱动MEIS1表达的多个增强器。总的来说，初步工作支持我 MEIS1是一种富含VSMC的转录因子的假设，可以通过 MyOCD的反式激活。我建议在此K99/R00奖的过程中以三个目标来检验这一假设。 K99 AIM 1将随着内源性MEIS1的损失阐明VSMC表型。 R00 AIM 2将阐明使用进化保存的MEIS1-响应元件对MyOCD体内的转录调节小鼠最先进的基因组编辑。 R00 AIM 3将阐明血管中MEIS1的功能性顺式调节体疾病。拟议研究的完成将为MEIS1对VSMC的贡献提供新的见解分化和血管稳态以及血管疾病中MEIS1的机械调节。在K99阶段，与多派相关研究有关的概念和实验培训将帮助我在我的R00阶段，追求高级MEIS1研究作为独立研究者，并为我定位良好我的第一个R01应用程序。