Glycation inactivation of human CD59 and diabetic complications

人 CD59 的糖基化失活与糖尿病并发症

基本信息

批准号：
8766558
负责人：
JOSE A HALPERIN
金额：
$ 41.01万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-12-15 至 2017-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8766558
关键词：
Accounting Active Sites Acute Adult Animal Model Animals Antibodies Aorta Apolipoprotein E Atherosclerosis Attenuated Backcrossings Biochemical Pathway Biological Response Modifiers Blood Glucose Blood Vessels Budgets Cardiovascular Diseases Cardiovascular system Cells Chronic Clinical Clinical Research Complement Complications of Diabetes Mellitus Deposition Development Diabetes Mellitus Diabetic Angiopathies Diabetic mouse Disease Endothelial Cells Endothelium Engineering Enzyme-Linked Immunosorbent Assay Epidemiologic Studies Erythrocytes Ethics Evans blue stain Exhibits Fatty acid glycerol esters Fructosamine Future Generations Genes Genetic Glucose Glycosylated Hemoglobin Goals Group Identifications Health Hemolytic Anemia Histologic Human Hyperglycemia Immune system Individual Inflammation Injection of therapeutic agent Insulin Insulin-Dependent Diabetes Mellitus Kidney Diseases Knock-out Laboratories Life Link Measurement Mediating Metabolic Pathway Modeling Molecular Morbidity - disease rate Mus Myocardial Ischemia Neuropathy Organ Pathogenesis Pathology Peripheral Vascular Diseases Plant Roots Plasma Play Population Prevention Prevention therapy Protein Isoforms Proteins Publications Publishing Relative (related person)Reporting Research Resistance Retinal Diseases Risk Role Severities Staining method Stains Streptozocin Stroke Testing Tissues Transgenic Mice United States National Institutes of Health VWF gene Vascular Diseases Work atherogenesis complement system diabetic diabetic cardiomyopathy follow-up genetic regulatory protein glycation mortality mouse model mutant novel preclinical study trait

项目摘要

DESCRIPTION (provided by applicant): Chronic high blood sugar levels (hyperglycemia) are responsible for diabetic complications including nephropathy, retinopathy, neuropathy and accelerated atherosclerosis, all major causes of morbidity and mortality in the adult population. However, how hyperglycemia causes diabetic complications is still poorly understood. Extensive clinical studies by us and others indicate that the complement system, a major effector of the immune system and mediator of inflammation, may play a significant role in the pathogenesis of these complications. In particular, we have established that human CD59, a key complement regulatory protein that protects cells from complement-mediated damage, is inhibited by glycation, the non-enzymatic attachment of glucose to proteins. This is because human CD59 contains a glycation motif, formed by its H44 residue at H 5A0 from its K41 residue in the active site of the protein. Interestingly, this glycation motif is not present in CD59 from other species. Our hypothesis is that glycation-inactivation of hCD59 due to its unique H44 residue could represent the elusive link between hyperglycemia and the vascular proliferative complications of human diabetes. However, practical and ethical considerations preclude conducting clinical studies to test this hypothesis comprehensively in humans. Therefore, our strategy to determine the cellular and molecular mechanism by which complement damages the target organs of diabetic complications has focused on developing molecular engineered mice that are now available in our laboratory. We will make these mice diabetic by STZ injection (type 1 diabetes model), and investigate the role of glycation-inactivation of hCD59 in diabetic cardiovascular disease taking advantage of our previous generation of mCD59KO mice backcrossed into the Apoe-/- background. Specifically, we will compare diabetic hCD59 wild type (glycation-inactivation sensitive) transgenic mice with diabetic hCD59Gln44 mutant (glycation- inactivation resistant) transgenic mice, both in a mCd59KO and Apoe-/-background. We will also use an available anti-mC5 antibody to block the terminal complement cascade and provide direct mechanistic evidence for the contribution of complement to diabetic vascular diseases. Successful accomplishment of this work will provide: 1) clear evidence for the role of complement and glycation-inactivation of hCD59 in the pathogenesis of the cardiovascular complications of diabetes; 2) needed animal models to study mechanism, therapy and prevention of diabetic complications; and 3) strong evidence to support future studies on complement and diabetes in humans.

描述（由申请人提供）：慢性高血糖水平（高血糖）是导致糖尿病并发症的原因，包括肾病、视网膜病、神经病和加速动脉粥样硬化，这些都是成年人发病和死亡的主要原因。然而，高血糖如何导致糖尿病并发症仍知之甚少。我们和其他人的广泛临床研究表明，补体系统是免疫系统的主要效应器和炎症介质，可能在这些并发症的发病机制中发挥重要作用。特别是，我们已经确定人类 CD59（一种关键的补体调节蛋白，可保护细胞免受补体介导的损伤）受到糖化（葡萄糖与蛋白质的非酶促附着）的抑制。这是因为人 CD59 含有糖基化基序，该基序由 H 5A0 处的 H44 残基与蛋白质活性位点的 K41 残基形成。有趣的是，这种糖基化基序不存在于其他物种的 CD59 中。我们的假设是，hCD59 由于其独特的 H44 残基而导致的糖基化失活可能代表高血糖与人类糖尿病血管增殖并发症之间难以捉摸的联系。然而，出于实际和伦理方面的考虑，无法进行临床研究来在人类身上全面检验这一假设。因此，我们确定补体损害糖尿病并发症靶器官的细胞和分子机制的策略重点是开发目前在我们实验室中可用的分子工程小鼠。我们将通过 STZ 注射使这些小鼠患糖尿病（1 型糖尿病模型），并利用我们上一代与 Apoe-/- 背景回交的 mCD59KO 小鼠，研究 hCD59 糖基化失活在糖尿病心血管疾病中的作用。具体来说，我们将在 mCd59KO 和 Apoe-/-背景中比较糖尿病 hCD59 野生型（糖化失活敏感）转基因小鼠与糖尿病 hCD59Gln44 突变体（糖化失活抗性）转基因小鼠。我们还将使用现有的抗 mC5 抗体来阻断末端补体级联，并为补体对糖尿病血管疾病的贡献提供直接的机制证据。这项工作的成功完成将提供：1）补体和hCD59糖化失活在糖尿病心血管并发症发病机制中的作用的明确证据； 2）研究糖尿病并发症发生机制、治疗和预防所需的动物模型； 3）强有力的证据支持未来对人类补体和糖尿病的研究。