Transcriptional Regulation by Angiotensin II in Vascular Smooth Muscle Cells

血管紧张素 II 在血管平滑肌细胞中的转录调节

• 批准号: 10458055
• 负责人: Zhen Bouman Chen
• 金额: $ 70.39万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10458055
• 关键词: 3-Dimensional; Address; Adult; Affect; Angiotensin II; Aorta; Arteries; Atherosclerosis; Bioinformatics; Biological Assay; Blood Vessels; CRISPR/Cas technology; Candidate Disease Gene; Cardiovascular Diseases; Cells; Chromatin; Clinical; Complications of Diabetes Mellitus; DNA; DNA Methylation; Data; Development; Diabetes Mellitus; Diabetic mouse; Enhancers; Epigenetic Process; Extracellular Matrix; Fibroblasts; Functional disorder; Funding; Genes; Genetic; Genome; Glucose; Growth Factor; Healthcare; Hi-C; Human; Hyperglycemia; Hypertension; In Situ; In Vitro; Inflammation; Inflammatory; Investigation; Knowledge; Lead; Losartan; Mediating; Memory; Metabolic; Methods; Molecular Conformation; Morbidity - disease rate; Mus; Patients; Pharmaceutical Preparations; Phenotype; Play; Production; RNA; Receptor, Angiotensin, Type 1; Role; Signal Transduction; Smooth Muscle Myocytes; Stimulus; Switch Genes; Synthetic Genes; Technology; Testing; Transcriptional Regulation; Type 2 diabetic; Untranslated RNA; Validation; Vascular Diseases; Vascular Smooth Muscle; XCL1 gene; cell behavior; cell type; diabetes management; diabetic; diabetic cardiomyopathy; epigenetic memory; epigenome; epigenomics; genome-wide; genomic locus; glycemic control; improved; in vivo; in vivo Model; inhibitor; innovation; insight; macrophage; methylome; migration; mortality; multiple omics; new therapeutic target; novel; novel therapeutics; programs; single cell sequencing; therapeutic target; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; translational potential; vascular smooth muscle cell proliferation

SUMMARY: Vascular smooth muscle cells (VSMCs) in the blood vessel wall play pivotal roles in cardiovascular disease (CVD; e.g., hypertension), which affects ~48% of US adults, and which is significantly accelerated by diabetes (DCVD). When stimulated by factors like diabetes and the growth factor Angiotensin II (AngII), mature VSMCs de-differentiate through “phenotypic switching” (Phe-sw) via dysregulation of "contractile" and "synthetic” genes, resulting in increased VSMC proliferation, migration, inflammation, and extracellular matrix (ECM) production. In the previous funding cycles, we unraveled the first functional roles for AngII-regulated long noncoding RNAs and (super-)enhancers (a key epigenetic regulatory layer) in VSMCs. Our overall objective in this renewal is to elucidate the mechanisms regulating diabetes- and AngII-induced VSMC Phe-sw at the epigenome and single-cell (sc) level and how these mechanisms establish vascular metabolic memory (in which prior hyperglycemia/AngII exposure leads to persistent long-term DCVD despite subsequent glucose normalization). We will use state-of-the-art multi-omics and sc-sequencing (seq) approaches to decipher VSMC behavior in DCVD and metabolic memory and identify new drug targets. Our hypothesis is that diabetic conditions and AngII coordinately re-program the VSMC transcriptome and epigenome, which lead to persistent dysregulation of genes promoting Phe-sw to unique cellular states underlying VSMC dysfunction and accelerated DCVD. This hypothesis is supported by extensive new preliminary data that show: i) a diabetic state augments AngII actions and promotes VSMC proliferation and Phe-sw gene profiles, which persist even after glucose normalization; ii) DNA methylation is decreased, and chromatin accessibility is increased at key upregulated ECM and inflammatory genes in VSMCs from diabetic mice, even after culture in normal glucose; iii) new cell clusters indicative of Phe-sw occur in aortas of Ang II-infused mice, identified using integrated scRNA- and scATAC-seq; iv) diabetic stimuli induce 3D chromatin changes in vascular cells, seen using chromatin conformation assays. We will test our hypothesis in 3 specific aims: 1) Define diabetes-induced transcriptomic and epigenomic changes [AngII-(in)dependent] in VSMCs at Phe-sw-related genes, and the memory of their persistent dysregulation after glucose normalization in vitro; 2) Elucidate the de-differentiated VSMC subtypes and their functions in diabetes- and AngII-induced Phe-sw in arteries, and their persistence after glucose normalization in vivo, using scRNA-seq and scATAC-seq; and 3) Determine the translational potential for reversing DCVD and vascular memory by targeting candidate genes/loci mediating diabetes- and AngII-induced VSMC Phe-sw. This innovative study, using cutting-edge technologies and functional in vivo models, will provide novel insights into VSMC regulatory networks and epigenetic memory of diabetic vasculopathy. This knowledge has the potential to inform development of much-needed new therapies, especially for patients not responding well to currently available diabetes drugs and AngII blockers, with far-reaching clinical implications for DCVD.

摘要：血管壁中的血管平滑肌细胞 (VSMC) 在心血管疾病中发挥着关键作用 疾病（CVD；例如高血压），影响约 48% 的美国成年人，并且显着加速 糖尿病（DCVD）。当受到糖尿病和生长因子血管紧张素 II (AngII) 等因素的刺激时，成熟 VSMC 通过“收缩”和“合成”失调的“表型转换”(Phe-sw) 去分化 基因，导致 VSMC 增殖、迁移、炎症和细胞外基质 (ECM) 增加 生产。在之前的融资周期中，我们阐明了 AngII 监管的长期基金的第一个功能角色。 VSMC 中的非编码 RNA 和（超级）增强子（关键的表观遗传调控层）。我们的总体目标是 这一更新是为了阐明调节糖尿病和 AngII 诱导的 VSMC Phe-sw 的机制 表观基因组和单细胞（sc）水平以及这些机制如何建立血管代谢记忆（其中 尽管随后出现血糖升高，但先前的高血糖/AngII 暴露仍会导致持续性长期 DCVD 正常化）。我们将使用最先进的多组学和 SC 测序 (seq) 方法来破译 VSMC DCVD 和代谢记忆的行为并确定新的药物靶点。我们的假设是糖尿病 条件和 AngII 协调重新编程 VSMC 转录组和表观基因组，从而导致持久的 基因失调促进 Phe-sw 进入 VSMC 功能障碍的独特细胞状态并加速 CVD。这一假设得到了广泛的新初步数据的支持，这些数据表明：i）糖尿病状态会增强 AngII 作用并促进 VSMC 增殖和 Phe-sw 基因谱，即使在葡萄糖后仍持续存在 正常化; ii) DN​​A 甲基化减少，关键上调的染色质可及性增加 糖尿病小鼠 VSMC 中的 ECM 和炎症基因，即使在正常葡萄糖培养后也是如此； iii) 新细胞 指示 Phe-sw 的簇出现在 Ang II 输注小鼠的主动脉中，使用整合的 scRNA 和 scATAC-seq； iv) 糖尿病刺激诱导血管细胞中的 3D 染色质变化，通过染色质观察到 构象测定。我们将在 3 个具体目标上检验我们的假设：1) 定义糖尿病诱导的转录组学 VSMC 中 Phe-sw 相关基因的表观基因组变化[AngII-(in)依赖性]，以及它们的记忆 体外血糖正常化后持续失调； 2) 阐明去分化的VSMC亚型 及其在糖尿病和 AngII 诱导的动脉中 Phe-sw 中的功能，以及葡萄糖后其持久性 使用 scRNA-seq 和 scATAC-seq 进行体内标准化； 3) 确定转化潜力 通过靶向介导糖尿病和血管紧张素II诱导的候选基因/位点来逆转 DCVD 和血管记忆 VSMC Phe-sw。这项创新研究利用尖端技术和功能性体内模型，将提供 对 VSMC 调节网络和糖尿病血管病变表观遗传记忆的新见解。这些知识 有潜力为急需的新疗法的开发提供信息，特别是对于没有反应的患者 与目前可用的糖尿病药物和 AngII 阻滞剂相比，对 DCVD 具有深远的临床意义。