Excess O-GlcNAc modification of proteins and myocardial fibrosis

蛋白质的过量 O-GlcNAc 修饰与心肌纤维化

基本信息

批准号：
10265339
负责人：
Francisco J Villarreal
金额：
--
依托单位：
VA SAN DIEGO HEALTHCARE SYSTEM
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-01-01 至 2022-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10265339
关键词：
Address Adenoviruses Adverse effects Age Age-Years Aging Amino Acids Animals Antipsychotic Agents Apoptosis Back Biochemical Biological Process Cardiac Cardiac development Cells Code Collagen Cytoplasmic Protein Cytoskeletal Modeling Data Development Diabetes Mellitus Disease Enzyme Inhibitor Drugs Excision Exposure to Female Fibroblasts Fibrosis Functional disorder Genetic Transcription Glucose Health Heart Heart Diseases Heart failure Hexosamines In Vitro Link Mediating Modeling Modification Molecular Monosaccharides Mutate Myocardial Myofibroblast N acetylglucosaminidase N-Acetylglucosaminyltransferases Non-Insulin-Dependent Diabetes Mellitus Nuclear Nuclear Proteins Organ Pathway interactions Patients Phenotype Play Population Post-Translational Protein Processing Production Proteins Proteomics Recombinants Reporting Research Risk Factors Role Secondary to Serine Signal Transduction Sp1 Transcription Factor Structure Testing Threonine Time Tissues Transgenic Mice Translations Veterans Work aged arginase cardiogenesis cationic antimicrobial protein CAP 37 coronary fibrosis diabetic enzyme activity glomerulosclerosis in vivo male military veteran mouse model non-diabetic novel novel strategies novel therapeutic intervention nucleocytoplasmic transport overexpression peptide O-linked N-acetylglucosamine-beta-N-acetylglucosaminidase prevent programs statistics

项目摘要

The major hypothesis to be tested in this proposal is that, “Reducing the excess addition of β-N- acetylglucosamine (O-GlcNAc) to regulatory factors present in cardiac fibroblasts ameliorates diabetes mellitus induced myocardial fibrosis”. Given the older average age of the Veteran population, about 25% suffer from type 2 diabetes mellitus (DM2). A large number of these patients will develop DM2 induced cardiac fibrosis, which adversely impacts diastolic function and frequently leads to the development of heart failure. Excess O- GlcNAc modification of proteins is known to occur with aging and most notably in the setting of DM2. We have demonstrated that excess O-GlcNAcylation of cardiac fibroblast (CF) proteins is associated with the enhanced production of collagens. As tissue fibrosis (e.g. glomerulosclerosis) is a prominent feature of DM2 our research findings may also have broader implications as a strategy to ameliorate excess collagen production by fibroblasts present in other organs. The post-translational modification of serine and threonine residues of nuclear and cytoplasmic proteins by the O-linked attachment of the monosaccharide β-N-acetylglucosamine is a highly dynamic and ubiquitous protein modification that is secondary to the action of β-N- acetylglucosaminyltransferase (OGT). Conversely, the removal of O-GlcNAc is mediated by N- acetylglucosaminidase (O-GlcNAcase). Protein O-GlcNAcylation is rapidly emerging as a key regulator of critical biological processes including nuclear transport, translation and transcription, signal transduction, cytoskeletal reorganization, proteasomal degradation, and apoptosis. We demonstrated that in high glucose treated CF, the nuclear transcription factor Sp1 and arginase evidence excess O-GlcNAcylation. Both of these proteins are intricately associated with stimulating the production of collagens. Expression in CF of an adenovirus coding for O-GlcNAcase, decreased Sp1 and arginase O-GlcNAcylation and restores high glucose- induced excess collagen production back to normal levels. However, no studies have identified which specific amino acid residues can be modified by O-GlcNAcylation and how they alter Sp1 and arginase function. Furthermore, these observations have not been evidenced in the in vivo setting and linked with changes in cardiac structure and function. Given these facts and the preliminary data we have generated, we propose to examine the following specific aims: Aim 1. To identify the development of myocardial fibrosis, diastolic dysfunction, Sp1 and arginase I residue modification in an aged model of DM2. Aim 2. To characterize HG induced CF amino acid residue O-GlcNAc modification of Sp1 and arginase I and its functional implications. Aim 3. To characterize the capacity of altered O-GlcNAcase activity to modify DM2 or HG induced alterations in Sp1 and arginase I residue modification, fibroblast/myofibroblast phenotype, cardiac structure and function.

该提议中要检验的主要假设是，“减少了β-n-的过量添加乙酰葡萄糖（O-GLCNAC）对心脏成纤维细胞中存在的调节因子可改善糖尿病诱发心肌纤维化”。鉴于老兵的年龄较大，大约25％的人患有 2型糖尿病（DM2）。这些患者中有大量会出现DM2诱导的心脏纤维化，这会对舒张功能产生不利影响，并经常导致心力衰竭的发展。多余的O- 已知蛋白质的GlcNAC修饰是随着衰老而发生的，最著名的是在DM2的情况下发生。我们有证明心脏成纤维细胞（CF）蛋白的O-Glcnacylation与增强生产胶原蛋白。由于组织纤维化（例如肾小球硬化）是DM2的重要特征。调查结果也可能具有更广泛的含义，以减轻通过其他器官中存在的成纤维细胞。丝氨酸和苏氨酸保留的翻译后修饰单糖β-N-乙酰葡萄糖的O连接附着的核和细胞质蛋白是高度动态和普遍存在的蛋白质修饰，是β-n-作用的继发的乙酰氨基氨基链酰酯转移酶（OGT）。相反，去除O-GlCNAC是由N-介导的乙酰葡萄糖氨基酶（O-GlcNACASE）。蛋白O-Glcnacylation迅速成为一个关键调节剂关键的生物学过程，包括核转运，翻译和转录，信号转导，细胞骨架重组，蛋白酶体降解和凋亡。我们证明了在高葡萄糖中处理过的CF，核转录因子SP1和精氨酸酶证据超过O-Glcnacylation。这两个蛋白质与刺激胶原蛋白的产生相关。在CF中的表达编码O-Glcnacase的腺病毒，SP1和精氨酸酶O-Glcnacylation降低并恢复高葡萄糖 - 诱导过多的胶原蛋白产生回到正常水平。但是，尚未确定哪个特定氨基酸残基可以通过O-Glcnacylation以及它们如何改变SP1和精氨酸酶功能来改变。此外，这些观察结果尚未在体内环境中得到证明，并与变化有关心脏结构和功能。鉴于这些事实和我们生成的初步数据，我们建议检查以下特定目的：目标1。确定心肌纤维化的发展 DM2老化模型中的功能障碍，SP1和精氨酸酶I的住所修饰。目标2。表征HG 诱导的CF氨基酸保留O-GLCNAC对SP1和精氨酸酶I及其功能意义的修饰。目标3。表征改变O-Glcnacase活性改变DM2或HG诱导改变的能力在SP1和精氨酸酶I修饰中，成纤维细胞/成肌纤维细胞表型，心脏结构和功能。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

In Pursuit of Understanding the Role of Estrogens in Regulating Cardiac Structure and Function.

DOI：
10.1016/j.jacbts.2020.08.002
发表时间：
2020-09
期刊：
JACC. Basic to translational science
影响因子：
0
作者：
Villarreal F
通讯作者：
Villarreal F

Restorative potential of (-)-epicatechin in a rat model of Gulf War illness muscle atrophy and fatigue.

DOI：
10.1038/s41598-021-01093-w
发表时间：
2021-11-08
期刊：
Scientific reports
影响因子：
4.6
作者：
Ramirez-Sanchez I;Navarrete-Yañez V;Garate-Carrillo A;Lara-Hernandez M;Espinosa-Raya J;Moreno-Ulloa A;Gomez-Diaz B;Cedeño-Garcidueñas AL;Ceballos G;Villarreal F
通讯作者：
Villarreal F

Development of muscle atrophy and loss of function in a Gulf-War illness model: underlying mechanisms.

海湾战争疾病模型中肌肉萎缩和功能丧失的发展：潜在机制。

DOI：
10.1038/s41598-020-71486-w
发表时间：
2020
期刊：
Scientific reports
影响因子：
4.6
作者：
Ramirez-Sanchez,Israel;Navarrete-Yañez,Viridiana;Garate-Carrillo,Alejandra;Loredo,Maria;Lira-Romero,Esmeralda;Estrada-Mena,Javier;Campeau,Anaamika;Gonzalez,David;Carrillo-Terrazas,Marvic;Moreno-Ulloa,Aldo;Ceballos,Guillermo;Villarrea
通讯作者：
Villarrea

Multi-omics study identifies novel signatures of DNA/RNA, amino acid, peptide, and lipid metabolism by simulated diabetes on coronary endothelial cells.

DOI：
10.1038/s41598-022-16300-5
发表时间：
2022-07-14
期刊：
SCIENTIFIC REPORTS
影响因子：
4.6
作者：
Moreno-Ulloa, Aldo;Delgado-De la Herran, Hilda Carolina;Alvarez-Delgado, Carolina;Mendoza-Porras, Omar;Carballo-Castaneda, Rommel A.;Donis-Maturano, Luis;Villarreal, Francisco
通讯作者：
Villarreal, Francisco