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.

在美国和全球，血管疾病仍然是死亡的主要原因。已经提出了代谢 重编程和增加的6-磷酸葡萄糖脱氢酶（G6PD）活性和表达有助于 致命的血管增生性血管病的发病机理。此外，一些研究表明，患有丧失的人 功能G6PD（地中海或非洲）变体 - S188F（G6PDS188F; A型；严重缺陷）或N126D （G6PDN126D; A型；轻度缺陷）非nonyny单核苷酸多态性 - 具有较低的频率 冠状动脉疾病。但是，G6PD驱动的致病性和G6PD变体相关的保护机制 影响血管疾病仍然难以捉摸。因此，我们建议确定由新的潜在机制 在血管平滑肌细胞（VSMC）的核中发现的G6PD同工型，有助于致病性较大 动脉刚度和重塑。基于强烈支持初步结果，我们假设核G6PD 是表观遗传修饰剂的调节剂，是VSMC中的转录调节剂。因此，功能丧失 G6PD（S188F，N126D）变体阻止表观遗传组的适应不良修饰，从而减少了大动脉弹性和 肥胖/代谢综合征和气球伤害引起的重塑。我们将以三个特定的特定方式检验该假设 目标。在AIM 1中，我们将检验以下假设：核中G6PD和/或G6PD协调的氧化还原控制 表观遗传修饰剂（DNA甲基转移酶（DNMT）和DNA（TET）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DeDiventiation（促炎， - 脉络性和 - 增殖）在VSMC中的表型。在AIM 2中， 我们将确定功能丧失的G6PD变体是否从表观遗传修饰符中脱离以增加DNA 甲基化，抑制组蛋白3-赖氨酸三甲基化，并减少会导致适应不良的基因的转录 VSMCS的炎症， - 脉络性和增生剂）特性。在AIM 3中，我们将确定G6PD变体是否 大鼠表现出更少的不良适应性表观遗传（组蛋白3-赖氨酸三甲基化），并且发生较小的动脉 弹性（刚度）和血管重塑比喂高脂饮食的野生型大鼠（肥胖/代谢模型 综合征）或受到颈动脉气球伤害。功能收益和功能丧失研究的结果 该项目的直接影响G6PD对与致病性血管重塑相关的基因表达的直接影响 和大动脉僵硬，导致心力衰竭和死亡。我们预见了对血管生物学的两个重大影响： [1]迄今未知的G6PD依赖性亚细胞氧化还原直接与基本的连接 血管病理学中的转录力学和基因转录和[2]开发新疗法 靶向氧化还原信号，以减少大动脉刚度和重塑。