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计算，使伪影僵硬并预测不良心血管死亡率和发病率。到目前为止，没有疗法直接靶向血管钙化。现在有强有力的证据确定血管平滑肌细胞（VSMC）到“骨状”细胞中的成骨分化是关键的为了开发血管计算。我们和其他人展示了SMC衍生的Runx2（Runt- 相关的转录因子2）对于控制VSMC的成骨分化至关重要，VSMC诱导了VSMC的成骨分化。动脉粥样硬化，糖尿病和肾脏疾病的血管钙化。使用单细胞RNA测序（SCRNA-SEQ）分析，我们发现了一种新型的Runx2抑制器及其对SMC表型开关的影响动脉粥样硬化。具体而言，我们确定蛋白精氨酸甲基转移酶1（PRMT1）扮演在抑制RUNX2和调节SMC表型开关中的关键作用。 PRMT1是一个新兴的调节器然而，人类病理过程在血管钙化和动脉粥样硬化中的功能完全是未知。因此，我们有责任提供有关PRMT1如何充当新Runx2的其他证据血管表型开关和钙化调节中的抑制剂。引人入胜观察Runx2上调与PRMT1在中的下调之间的反相关性观察计算出的人和小鼠的动脉粥样硬化病变，我们使用PRMT1增益进行了功能研究以及功能丧失VSMC和我们的新型SMC特异性PRMT1转基因小鼠。我们的初步研究在体外，PRMT1在调节中表现出了pRMT1的致病作用； SMC特异性转基因PRMT1抑制了主动脉runx2和体内动脉粥样硬化的发展 apoe - / - 小鼠。进一步的证据表明，PRMT1指导的RUNX2调节是通过 Runx2的甲基化。 Runx2的无偏蛋白质组学分析相互作用揭示了Runx2的相互作用血清反应因子（SRF），这是收缩SMC标记基因的基本转录调节剂，该调节因子 SMC标记基因的SRF依赖性表达失调。基于这些新的令人兴奋的发现，我们假设PRMT1是一个键Runx2抑制器，它调节VSMC Runx2并控制VSMC 动脉粥样硬化中的表型转换和钙化。利用新的SMC特异性PRMT1转基因鼠标模型和全面的多词方法，该提案将发现一种新颖的调节在调节VSMC表型开关和钙化时突出显示PRMT1/RUNX2信号轴的范式。