Proteome of Glomerular Cells in Diabetic Nephropathy

糖尿病肾病肾小球细胞的蛋白质组

• 批准号: 6826587
• 负责人: PAUL A VOZIYAN
• 金额: $ 29.9万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6826587
• 关键词: cellular pathology; chronic renal failure; diabetic nephropathy; electrospray ionization mass spectrometry; extracellular matrix; glucose; glycation; high performance liquid chromatography; hyperglycemia; kidney cell; kidney pharmacology; matrix assisted laser desorption ionization; molecular pathology; pathologic process; podocyte; protein structure function; proteomics; renal glomerulus; tissue /cell culture; two dimensional gel electrophoresis; vitamin B6

DESCRIPTION (provided by applicant): The chronic hyperglycemia of diabetes leads to serious complications that include diabetic nephropathy, the leading cause of end-stage renal disease. It is largely unknown how the elevated levels of glucose in plasma and tissues are translated into renal pathology, which is characterized by the expansion of mesangial matrix and thickening of glomerular basement membrane (GBM). Clearly, the effects of hyperglycemia on glomerular cells that interact with mesangial matrix and GBM in kidney are at the heart of pathogenic mechanisms of diabetic nephropathy. One of the major consequences of hyperglycemic conditions is the acceleration of glycation reactions, i.e., modifications of proteins by glucose forming advanced glycation endproducts or AGEs. Thus, the elevated glucose levels through glucose modification of proteins could bring about changes in glomerular cell behavior. These cellular changes result in modulation of expression of vascular adhesion molecules and growth factors, increased excretion of extracellular matrix proteins, and induction of pro-inflammatory responses. To understand such a complex pathogenic mechanism, global changes that occur in the cell proteins require identification at the molecular level. With this knowledge, new protein targets could be identified to address pathogenic mechanisms and develop new therapies. An earlier work in our laboratory has discovered that pyridoxamine (PM) is an in vitro inhibitor of protein-AGE formation. In vivo, PM prevented development of early renal disease in animal models of diabetes and is currently in Phase II clinical trials for treatment of diabetic nephropathy. Mechanism of PM action is unknown but it is likely that PM interferes with pathogenic cellular effects of glycated proteins. New proteomic technologies that allow characterizing cellular proteome offer a powerful tool for determining molecular mechanisms of diabetic nephropathy. This proposal will employ such technologies to study cellular changes brought about by hyperglycemia and glucose-modified proteins. We hypothesize that glucose and glucose-modified proteins induce complex changes in the proteome of glomerular cells altering their interaction with extracellular matrix and leading to development of diabetic nephropathy. This central hypothesis will be addressed in four specific aims designed to determine 1) the changes in the proteome of glomerular cells cultured under normal and diabetic levels of glucose; 2) the effects of glycated proteins on cellular proteome; 3) the identities of glomerular cellular receptors that interact with glycated proteins; and 4) the effect of a prospective drug pyridoxamine on changes in the proteome of glomerular cells induced by glucose and glucose-modified proteins. The unbiased proteomic approach that interrogates a totality of cellular proteins for pathogenic changes is likely to result in a new knowledge about the mechanisms of diabetic nephropathy and to identify possible cellular protein targets for novel therapeutic treatments.

描述（由申请人提供）：糖尿病的慢性高血糖导致严重的并发症，包括糖尿病性肾病，这是终末期肾脏疾病的主要原因。血浆和组织中葡萄糖水平升高的水平在很大程度上被转化为肾脏病理学，其特征在于肾小球基质的扩展和肾小球基底膜（GBM）的增厚。 显然，高血糖对与肾脏中肾小球基质和GBM相互作用的肾小球细胞的影响是糖尿病性肾病致病机制的核心。 高血糖条件的主要后果之一是糖基化反应的加速度，即通过形成晚期糖基化终产物或年龄来修饰蛋白质的蛋白质。 因此，通过葡萄糖修饰蛋白质的葡萄糖水平升高可能会带来肾小球细胞行为的变化。 这些细胞变化导致调节血管粘附分子和生长因子的表达，细胞外基质蛋白的排泄增加以及促炎反应的诱导。为了理解这种复杂的致病机制，细胞蛋白中发生的全球变化需要在分子水平上进行鉴定。有了这些知识，可以确定新的蛋白质靶标，以解决致病机制并开发新的疗法。 我们实验室的早期工作发现吡id胺（PM）是蛋白质年龄形成的体外抑制剂。 在体内，PM阻止了糖尿病动物模型中肾脏早期疾病的发展，目前正在接受II期临床试验以治疗糖尿病性肾病。 PM作用的机理尚不清楚，但PM可能会干扰糖化蛋白的致病细胞作用。允许表征细胞蛋白质组的新蛋白质组学技术为确定糖尿病性肾病的分子机制提供了强大的工具。 该建议将采用此类技术研究高血糖和葡萄糖改性蛋白带来的细胞变化。 我们假设葡萄糖和葡萄糖改性的蛋白会诱导肾小球细胞蛋白质组的复杂变化，从而改变了它们与细胞外基质的相互作用并导致糖尿病性肾病的发展。 该中心假设将在四个旨在确定的特定目的中解决。1）在正常和糖尿病水平的葡萄糖水平下培养的肾小球细胞的蛋白质组变化； 2）糖化蛋白对细胞蛋白质组的影响； 3）与糖化蛋白相互作用的肾小球细胞受体的身份； 4）前瞻性药物吡啶胺对葡萄糖和葡萄糖改性蛋白诱导的肾小球细胞蛋白质组变化的影响。 质疑整体细胞蛋白以进行致病性变化的无偏蛋白质组学方法可能会导致有关糖尿病肾病机制的新知识，并确定新型治疗治疗的可能细胞蛋白靶标。