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.

该提案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）与心脏成纤维细胞中存在的调节因子相结合可改善糖尿病 鉴于退伍军人的平均年龄较高，大约 25% 的人患有 2 型糖尿病 (DM2) 大量患者会出现 DM2 诱发的心脏纤维化， 这会对舒张功能产生不利影响，并经常导致心力衰竭。 已知蛋白质的 GlcNAc 修饰会随着衰老而发生，并且在 DM2 的情况下最为显着。 证明心脏成纤维细胞 (CF) 蛋白的过量 O-GlcNAc 酰化与增强的 由于组织纤维化（例如肾小球硬化）是 DM2 的一个显着特征，因此我们的研究 研究结果还可能具有更广泛的影响，作为改善胶原蛋白过度生成的策略 成纤维细胞存在于其他器官中的丝氨酸和苏氨酸残基的翻译后修饰。 通过单糖 β-N-乙酰氨基葡萄糖的 O 连接形成核蛋白和细胞质蛋白 一种高度动态且普遍存在的蛋白质修饰，继发于 β-N- 的作用 乙酰氨基葡萄糖转移酶 (OGT) 离线时，O-GlcNAc 的去除是由 N-介导的。 乙酰氨基葡萄糖苷酶（O-GlcNAcase）正在迅速成为蛋白质 O-GlcNAc 的关键调节剂。 关键的生物过程，包括核运输、翻译和转录、信号转导、 我们证明了在高葡萄糖条件下的细胞骨架重组、蛋白酶体降解和细胞凋亡。 CF、核转录因子 Sp1 和精氨酸酶可以治疗过量的 O-GlcNAcylation。 蛋白质与刺激 CF 中胶原蛋白的产生密切相关。 编码 O-GlcNAcase 的腺病毒，减少 Sp1 和精氨酸酶 O-GlcNA 酰化并恢复高葡萄糖- 诱导过量的胶原蛋白生成恢复到正常水平但是，没有研究确定具体是哪一种。 氨基酸残基可以通过 O-GlcNAcylation 进行修饰，以及它们如何改变 Sp1 和精氨酸酶功能。 此外，这些观察结果尚未在体内环境中得到证实，并且与 鉴于这些事实和我们生成的初步数据，我们建议： 检查以下具体目标： 目标 1. 确定心肌纤维化、舒张期的发展 DM2 衰老模型中的功能障碍、Sp1 和精氨酸酶 I 残基修饰 目标 2. 表征 HG。 诱导 CF 氨基酸残基 O-GlcNAc 对 Sp1 和精氨酸酶 I 的修饰及其功能意义。 目标 3. 表征改变的 O-GlcNAcase 活性修改 DM2 或 HG 诱导的改变的能力 Sp1 和精氨酸酶 I 残基修饰、成纤维细胞/肌成纤维细胞表型、心脏结构和功能。

项目成果

Beneficial Effects of Flavonoids on Skeletal Muscle Health: A Systematic Review and Meta-Analysis.

黄酮类化合物对骨骼肌健康的有益作用：系统评价和荟萃分析。

• DOI: 10.1089/jmf.2021.0054
• 发表时间: 2022-04-08
• 期刊: Journal of medicinal food
• 影响因子: 2.4
• 作者: L. Munguía;Miguel Ortiz;Cristian González;Andrés Portilla;E. Meaney;F. Villarreal;Nayelli Nájera;G. Ceballos
• 通讯作者: G. Ceballos

The role of inflammation in driving left ventricular remodeling in a pre-HFpEF model.

炎症在 HFpEF 前模型中驱动左心室重塑的作用。

• 发表时间: 2020
• 作者: Loredo;Ramirez;Bustamante;Ayala, Marcos;Navarrete, Viridiana;Garate;Ito, Bruce R;Ceballos, Guillermo;Omens, Jeffrey;Villarreal, Francisco
• 通讯作者: Villarreal, Francisco

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

多组学研究通过模拟糖尿病对冠状动脉内皮细胞识别 DNA/RNA、氨基酸、肽和脂质代谢的新特征。

• 发表时间: 2022-07-14
• 影响因子: 4.6
• 作者: Moreno;Delgado;Álvarez;Mendoza;Carballo;Donis;Villarreal, Francisco
• 通讯作者: Villarreal, Francisco

Is it possible to treat nonalcoholic liver disease using a flavanol-based nutraceutical approach? Basic and clinical data.

是否有可能使用基于黄烷醇的营养保健方法治疗非酒精性肝病？

• 发表时间: 2022-11-01
• 作者: Hidalgo, Isabel;Ortiz;Villarreal, Francisco;Fonseca;Ceballos, Guillermo;Meaney, Eduardo;Nájera, Nayelli
• 通讯作者: Nájera, Nayelli

Antifibrotic Effects of (-)-Epicatechin on High Glucose Stimulated Cardiac Fibroblasts.

(-)-表儿茶素对高血糖刺激的心脏成纤维细胞的抗纤维化作用。

• DOI: 10.1089/jmf.2020.0210
• 发表时间: 2021-07-05
• 期刊: Journal of medicinal food
• 影响因子: 2.4
• 作者: Alej;ra Garate;ra;I. Ramírez‐Sánchez;Justina P. Nguyen;Julisa Gonzalez;G. Ceballos;F. Villarreal
• 通讯作者: F. Villarreal