Protein Arginine Methylation in Vascular Smooth Muscle Cell Phenotypic Modulation and Calcification

血管平滑肌细胞表型调节和钙化中的蛋白质精氨酸甲基化

基本信息

批准号：
10734531
负责人：
Yabing Chen
金额：
$ 71.4万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
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 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10734531
关键词：
Aging Aorta Apolipoprotein E Arginine Arterial Fatty Streak Atherosclerosis Blood Vessels Calcium Cardiovascular system Cells ChIP-seq Chronic Kidney Failure Clinical Research Complex DNA Binding Deposition Development Diabetes Mellitus Diet Disease Down-Regulation Elasticity Genes Genetic Transcription Health Homeostasis Human In Vitro Kidney Diseases Link Lipids Malignant Neoplasms Mediating Methylation Molecular Morbidity - disease rate Mus Oxidative Stress Induction Pathogenesis Pathologic Pathologic Processes Phenotype Play Protein-Arginine N-Methyltransferase Proteins Proteomics Regulation Role Serum Response Factor Signal Transduction Smooth Muscle Myocytes Testing Transgenic Mice Transgenic Organisms Up-Regulation Vascular Diseases Vascular Smooth Muscle Vascular Smooth Muscle Tissue Vascular calcification acute myeloid leukemia 1 protein arterial stiffness bone calcification design experimental study gain of function human tissue improved in vivo loss of function mortality mouse model multiple omics myocardin novel osteogenic protein arginine methyltransferase 2 single-cell RNA sequencing therapy development transcription factor transcriptomics

项目摘要

Abstract Vascular calcification in blood vessels, stiffens artery and predicts adverse cardiovascular mortality and morbidity. No therapies developed so far directly targeting vascular calcification. Strong evidence has now determined the osteogenic differentiation of vascular smooth muscle cells (VSMC) into “bone-like” cells is critical for the development of vascular calcification. We and others have demonstrated SMC-derived Runx2 (Runt- related transcriptional factor 2) is essential in regulating osteogenic differentiation of VSMC, which induces vascular calcification in atherosclerosis, diabetes and kidney disease. Using single cell RNA sequencing (scRNA-seq) analysis, we discovered a novel Runx2 suppressor and its impact on SMC phenotypic switch in atherosclerosis. Specifically, we determined that the protein arginine methyltransferase 1 (PRMT1) plays a critical role in inhibiting Runx2 and modulating SMC phenotypic switch. PRMT1 is an emerging regulator in human pathological processes, however, its function in vascular calcification and atherosclerosis is entirely unknown. It is thus incumbent upon us to provide additional evidence as to how PRMT1 acts as a new Runx2 suppressor in the modulation of vascular phenotypic switch and calcification. Prompted by the intriguing observations of an inverse correlation between upregulation of Runx2 and marked downregulation of PRMT1 in the calcified atherosclerotic lesions in human and mice, we carried out functional studies using the PRMT1 gain- and loss-of-function VSMC and our novel SMC-specific PRMT1 transgenic mice. Our preliminary studies demonstrated a causative role of PRMT1 in regulating Runx2, SMC phenotypic switch and calcification in vitro; and SMC-specific transgenic PRMT1 inhibited aortic Runx2 and the development of atherosclerosis in vivo in the ApoE-/- mice. Further evidence implicates that PRMT1-directed regulation of Runx2 is mediated through methylation of Runx2. Unbiased proteomics analysis of Runx2 interactome uncovered the interaction of Runx2 with serum response factor (SRF), an essential transcriptional regulator for contractile SMC marker genes, which dysregulates SRF-dependent expression of SMC marker genes. Based on these new and exciting findings, we hypothesize that PRMT1 is a key Runx2 suppressor, which regulates VSMC Runx2 and governs VSMC phenotypic switch and calcification in atherosclerosis. Utilizing the new SMC-specific PRMT1 transgenic mouse model and comprehensive multi-Omics approaches, the proposal will uncover a novel regulatory paradigm highlighting the PRMT1/Runx2 signaling axis in modulating VSMC phenotypic switch and calcification.

抽象的血管中的血管钙化、动脉硬化并预测不良心血管死亡率和迄今为止，尚未开发出直接针对血管钙化的疗法。确定血管平滑肌细胞（VSMC）成骨分化为“骨样”细胞至关重要我们和其他人已经证明了 SMC 衍生的 Runx2（Runt-）。相关转录因子 2）对于调节 VSMC 的成骨分化至关重要，从而诱导使用单细胞 RNA 测序研究动脉粥样硬化、糖尿病和肾脏疾病中的血管钙化。 (scRNA-seq) 分析中，我们发现了一种新型 Runx2 抑制因子及其对 SMC 表型转换的影响具体来说，我们确定蛋白质精氨酸甲基转移酶 1 (PRMT1) 在动脉粥样硬化中发挥着重要作用。 PRMT1 是一种新兴的调节因子，在抑制 Runx2 和调节 SMC 表型转换中发挥关键作用。然而，它在血管钙化和动脉粥样硬化中的作用完全是人类的病理过程。因此，我们有责任提供更多证据来证明 PRMT1 如何充当新的 Runx2。血管表型转换和钙化调节中的抑制因子由有趣的提示引起。观察到 Runx2 上调与 PRMT1 显着下调之间存在负相关关系针对人类和小鼠的钙化动脉粥样硬化病变，我们使用 PRMT1 增益进行了功能研究和功能丧失的 VSMC 以及我们的新型 SMC 特异性 PRMT1 转基因小鼠。证明了 PRMT1 在体外调节 Runx2、SMC 表型转换和钙化中的因果作用； SMC特异性转基因PRMT1抑制主动脉Runx2和体内动脉粥样硬化的发展 ApoE-/- 小鼠的进一步证据表明，PRMT1 介导的 Runx2 调节。 Runx2 相互作用组的无偏蛋白质组学分析揭示了 Runx2 的相互作用。血清反应因子（SRF）是收缩性 SMC 标记基因的重要转录调节因子，基于这些令人兴奋的新发现，我们发现 SRF 依赖性 SMC 标记基因的表达失调。 PRMT1 是一个关键的 Runx2 抑制因子，它调节 VSMC Runx2 并控制 VSMC 利用新的 SMC 特异性 PRMT1 转基因治疗动脉粥样硬化的表型转换和钙化。小鼠模型和综合多组学方法，该提案将揭示一种新的监管方法该范例强调了 PRMT1/Runx2 信号轴在调节 VSMC 表型转换和钙化中的作用。