Glycation inactivation of human CD59 and diabetic complications

人 CD59 的糖基化失活与糖尿病并发症

• 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
• 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; 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

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）：慢性血糖水平（高血糖）负责糖尿病并发症，包括肾病，视网膜病，神经病和加速性动脉粥样硬化，这是成人人群发病和死亡的所有主要原因。但是，高血糖导致糖尿病并发症的高血糖量仍然很少了解。我们和其他人的广泛临床研究表明，补体系统是免疫系统和炎症介体的主要效应因子，可能在这些并发症的发病机理中起重要作用。特别是，我们已经确定人CD59是一种保护细胞免受补体介导的损伤的关键补体调节蛋白，受到糖基化的抑制，即葡萄糖与蛋白质的非酶附着。这是因为人CD59包含一个糖基序，其由其H44残基在蛋白质活性位点的K41残基处形成H44残基。有趣的是，其他物种的CD59中不存在这种糖化基序。我们的假设是，由于其独特的H44残基，HCD59的糖基化灭活可能代表了高血糖与人类糖尿病的血管增殖并发症之间的难以捉摸的联系。然而，实践和道德考虑因素排除进行临床研究以全面检验该假设。因此，我们确定补体损害糖尿病并发症目标器官的细胞和分子机制的策略集中在开发我们实验室中现在可用的分子工程小鼠。我们将通过STZ注射（1型糖尿病模型）使这些小鼠糖尿病，并研究HCD59在糖尿病性心血管疾病中的糖基化糖化作用，利用了我们上一代的MCD59KO小鼠，回到了APOE-/ - 背景中。具体而言，我们将在MCD59KO和APOE-/apoE-/apoE-/apoE-/apoE-/ - / - / - 与糖尿病性HCD59野生型（糖化灭活敏感性）转基因小鼠中进行比较，与MCD59KO和APOE-/APOE-/ - / - / - / - - / - - / - - / - - / - - / - - / - - / - - / - - / - - / - - / - - / - - / - - / - - / - 。我们还将使用可用的抗MC5抗体来阻断末端补体级联反应，并为补体对糖尿病血管疾病的贡献提供直接的机械证据。这项工作的成功完成将提供：1）HCD59在糖尿病心血管并发症的发病机理中的补体和糖基化诱导作用的明确证据； 2）需要动物模型来研究糖尿病并发症的机制，治疗和预防； 3）有力的证据支持未来关于人类补体和糖尿病的研究。