Regulation of Vascular Smooth Muscle Cell Phenotype by a Novel Isoform of Glucose-6-Phosphate Dehydrogenase

新型葡萄糖-6-磷酸脱氢酶异构体对血管平滑肌细胞表型的调节

• 批准号: 10561265
• 负责人: SACHIN A GUPTE
• 金额: $ 70.55万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-15 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10561265
• 关键词: ATAC-seq; Affect; African; Arteries; Biology; Blood Vessels; Cardiovascular system; Carotid Arteries; Cause of Death; Cell Nucleus; Cessation of life; Chromatin; Coronary Arteriosclerosis; Coronary artery; DNA; DNA Binding; DNA Methylation; DNA Modification Methylases; DNase-I Footprinting; Development; Disease; Enzymes; Epigenetic Process; Evolution; Experimental Designs; FAIRE sequencing; Frequencies; Gene Expression; Gene Expression Profile; Genes; Genetic Polymorphism; Genetic Transcription; Glucosephosphate Dehydrogenase; Goals; Growth; Heart failure; High Fat Diet; Histones; Human; Hypertension; Individual; Inflammatory; Injury; Knowledge; Laboratories; Life; Link; Lysine; Mechanics; Medicine; Metabolic; Metabolic syndrome; Metabolism; Methylation; Modeling; Modernization; Modification; Molecular Weight; NADP; Nuclear; Obesity; Oxidation-Reduction; Pathogenesis; Pathogenicity; Pathology; Phenotype; Predisposition; Property; Protein Isoforms; Proteins; Pulmonary artery structure; Rattus; Regulation; Repression; Role; Signal Transduction; Single Nucleotide Polymorphism; Smooth Muscle Myocytes; Switch Genes; Technology; Testing; Variant; Vascular Diseases; Vascular Smooth Muscle; Vascular remodeling; arterial stiffness; design; epigenome; epigenomics; gain of function; glucose metabolism; histone methylation; loss of function; metabolomics; migration; myocardin; new therapeutic target; novel; prevent; prime editing; promoter; thrombotic; transcription factor

Vascular diseases continue to be a major cause of death in the US and worldwide. It has been proposed that metabolic reprogramming and increased glucose-6-phosphate dehydrogenase (G6PD) activity and expression contribute to the pathogenesis of fatal angioproliferative vasculopathies. Moreover, some studies suggest individuals with a loss-of- function G6PD (Mediterranean or African) variant – S188F (G6PDS188F; Type A-; severe deficiency) or N126D (G6PDN126D; Type A; mild deficiency) nonsynonymous single nucleotide polymorphism – have lower frequencies of coronary artery disease. However, G6PD-driven pathogenic and G6PD variant-associated protective mechanisms affecting vascular diseases remain elusive. We therefore propose to determine potential mechanisms, driven by a newly discovered G6PD isoform in the nucleus of vascular smooth muscle cells (VSMCs), that contribute to pathogenic large artery stiffness and remodeling. Based on strongly supporting preliminary results, we hypothesized that nuclear G6PD is a modulator of epigenetic modifiers and is a transcription regulator in VSMCs. Consequently, the loss-of-function G6PD (S188F, N126D) variants block maladaptive modifications of the epigenome, reducing large artery elastance and remodeling elicited by obesity/metabolic syndrome and balloon-injury. We will test this hypothesis in three specific aims. In Aim 1, we will test the hypothesis that G6PD and/or G6PD-coordinated redox in the nucleus controls the expression and activity of epigenetic modifiers (DNA methyltransferases (DNMT) and DNA (TET) and histone (JARID) demethylases) and transcription of genes that encode proteins involved in regulating the differentiation (contractile) and dedifferentiation (pro-inflammatory, -thrombotic, and -proliferative) phenotypes in VSMCs. In Aim 2, we will determine whether loss-of-function G6PD variants detach from epigenetic modifiers to increase DNA methylation, suppress histone3-lysine4 trimethylation, and reduce transcription of genes that confer maladaptive (pro- inflammatory, -thrombotic, and -proliferative) properties to VSMCs. In Aim 3, we will determine whether G6PD variant rats express fewer maladaptive epigenetic (histone3-lysine4 trimethylation) changes and develop less large artery elastance (stiffness) and vascular remodeling than wild-type rats fed a high-fat diet (a model of obesity/metabolic syndrome) or subjected to carotid artery balloon-injury. The results from gain-of-function and loss-of-function studies of this project will reveal the direct effect of G6PD on gene expression associated with pathogenic vascular remodeling and large artery stiffness, which lead to heart failure and death. We foresee two significant impacts on vascular biology: [1] linkage of heretofore unknown G6PD-dependent subcellular redox in the nucleus directly to the fundamental transcriptional mechanics and gene transcription in vascular pathobiology and [2] development of new treatments targeting redox signaling to reduce large artery stiffness and remodeling.

血管疾病仍然是美国和世界范围内的主要死亡原因。有人提出，代谢 重编程和增加的葡萄糖-6-磷酸脱氢酶（G6 PD）活性和表达有助于 致命性血管增生性血管病的发病机制。此外，一些研究表明，个人的损失- 功能G6 PD（地中海或非洲）变体-S188 F（G6 PDS 188 F; A型-;严重缺乏）或N126 D （G6 PDN 126 D; A型;轻度缺乏）非同义单核苷酸多态性-具有较低的 冠心病然而，G6 PD驱动的致病性和G6 PD变体相关的保护机制， 影响血管疾病仍然是难以捉摸的。因此，我们建议确定潜在的机制，由一个新的 在血管平滑肌细胞（VSMCs）的细胞核中发现了G6 PD亚型，这有助于致病性大 动脉僵硬和重塑。基于强有力的支持初步结果，我们假设核G6 PD 是表观遗传修饰物的调节剂，并且是VSMC中的转录调节因子。因此，功能丧失 G6 PD（S188 F，N126 D）变体阻断表观基因组的适应不良修饰，降低大动脉弹性， 由肥胖/代谢综合征和球囊损伤引起的重塑。我们将在三个具体的实验中验证这一假设。 目标。在目的1中，我们将检验细胞核中G6 PD和/或G6 PD-协调的氧化还原控制细胞核中的G6 PD和/或G6 PD-协调的氧化还原的假设。 表观遗传修饰物（DNA甲基转移酶（DNMT）和DNA（泰特）和组蛋白）的表达和活性 （JARID）脱甲基酶）和编码参与调节分化的蛋白质的基因的转录。 （收缩）和去分化（促炎、血栓形成和增殖）表型。在目标2中， 我们将确定功能丧失的G6 PD变体是否与表观遗传修饰剂分离，以增加DNA 甲基化，抑制组蛋白3-赖氨酸4三甲基化，并减少基因的转录，赋予适应不良（前 炎性、血栓形成和增殖）特性。在目标3中，我们将确定G6 PD变体 大鼠表现出较少的适应不良表观遗传学（组蛋白3-赖氨酸4三甲基化）变化，并形成较少的大动脉 弹性（僵硬）和血管重塑比野生型大鼠喂养高脂肪饮食（肥胖/代谢模型 综合征）或颈动脉球囊损伤。功能获得和功能丧失研究的结果 本项目的研究将揭示G6 PD对致病性血管重塑相关基因表达的直接影响 和大动脉僵硬，导致心力衰竭和死亡。我们预见到对血管生物学的两个重大影响： [1]细胞核中迄今未知的G6 PD依赖性亚细胞氧化还原直接与细胞核中的基本 血管病理生物学中的转录机制和基因转录以及[2]新治疗方法的开发 靶向氧化还原信号传导以减少大动脉僵硬和重塑。