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

• 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
• 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

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.

这项建议中要检验的主要假设是，“减少β-N的过量添加-- 乙酰氨基葡萄糖(O-GlcNAc)对心脏成纤维细胞调节因子的作用 诱导心肌纤维化“。考虑到退伍军人的平均年龄较大，约25%的人患有 2型糖尿病(DM2)。这些患者中的大量会发展成DM2诱导的心脏纤维化， 这会对舒张期功能产生不利影响，并经常导致心力衰竭的发展。过量的O- 已知蛋白质的GlcNAc修饰随着年龄的增长而发生，最显著的是在DM2的环境中发生。我们有 证明心脏成纤维细胞(CF)蛋白的过量O-GlcN酰化与增强 胶原蛋白的生产。由于组织纤维化(如肾小球硬化)是DM2的一个显著特征，我们的研究 研究结果可能还具有更广泛的意义，可以作为一种策略，通过以下方式改善过剩的胶原蛋白产生 成纤维细胞存在于其他器官中。丝氨酸和苏氨酸残基的翻译后修饰 核质蛋白通过单糖β-N-乙酰氨基葡萄糖的O-键连接 一种高度动态和普遍存在的蛋白质修饰，仅次于β-N- 乙酰氨基葡萄糖转移酶(OGT)。相反，O-GlcNAc的去除是由N-介导的。 乙酰氨基葡萄糖苷酶(O-GlcNAcase)。蛋白质O-GlcN酰化正在迅速崛起，成为一种关键的 关键的生物学过程，包括核运输、翻译和转录、信号转导、 细胞骨架重组、蛋白酶体降解和细胞凋亡。我们证明了在高糖环境下 处理后的CF核转录因子Sp1和精氨酸酶表现出过量的O-GlcN酰化。这两个都是 蛋白质与刺激胶原蛋白的产生有着错综复杂的联系。An在Cf中的表达 编码O-GlcNAcase的腺病毒降低Sp1和精氨酸酶O-GlcN酰化并恢复高血糖- 诱导过多的胶原蛋白产生恢复到正常水平。然而，还没有研究确定哪种特定的 氨基酸残基可以通过O-GlcN酰化修饰，以及它们如何改变Sp1和精氨酸酶的功能。 此外，这些观察结果还没有在活体环境中得到证实，并与 心脏的结构和功能。鉴于这些事实和我们产生的初步数据，我们建议 检查以下具体目标：目的1.确定心肌纤维化的发展，舒张期 老年DM2模型中的功能障碍、Sp1和精氨酸酶I残基修饰。目的2.确定HG的特征 Sp1和精氨酸酶I的Cf氨基酸残基O-GlcNAc修饰及其功能意义 目的3.研究O-GlcNAcase活性改变对DM2或HG诱导的改变的影响 在Sp1和精氨酸酶I残基修饰中，成纤维细胞/肌成纤维细胞表型、心脏结构和功能。

项目成果

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

Sex related differences in the pathogenesis of organ fibrosis.

• DOI: 10.1016/j.trsl.2020.03.008
• 发表时间: 2020-08
• 期刊: Translational research : the journal of laboratory and clinical medicine
• 作者: Garate-Carrillo A;Gonzalez J;Ceballos G;Ramirez-Sanchez I;Villarreal F
• 通讯作者: Villarreal F

Altered Myokine Secretion Is an Intrinsic Property of Skeletal Muscle in Type 2 Diabetes.

• DOI: 10.1371/journal.pone.0158209
• 发表时间: 2016
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Ciaraldi TP;Ryan AJ;Mudaliar SR;Henry RR
• 通讯作者: Henry RR

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