Probing the Molecular Basis of Glomerular Injury in Diabetic Nephropathy

探讨糖尿病肾病肾小球损伤的分子基础

• 批准号: 8593948
• 负责人: Megan Murray Gessel
• 金额: $ 3.93万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8593948
• 关键词: Address; Advanced Glycosylation End Products; Affect; Basement membrane; Biochemical; Cells; Chemistry; Chronic; Clinical; Complex; Complications of Diabetes Mellitus; Development; Diabetes Mellitus; Diabetic Nephropathy; Dialysis procedure; Disease; Early Diagnosis; End stage renal failure; Event; Extracellular Matrix; Extracellular Protein; Fellowship; Fibrosis; Goals; Hyperglycemia; Hypertrophy; Image; In Situ; In Vitro; Injury; Institution; Kidney; Kidney Diseases; Kidney Failure; Kidney Transplantation; Lead; Life; Lipids; Mass Spectrum Analysis; Measures; Methods; Modification; Molecular; Molecular Probes; Monitor; Morbidity - disease rate; Mus; Onset of illness; Pathogenesis; Pathology; Pathway interactions; Patients; Peptides; Play; Post-Translational Protein Processing; Proteins; Protocols documentation; Pyridoxamine; Reaction; Reactive Oxygen Species; Reporting; Research; Research Personnel; Research Proposals; Research Training; Role; Scientist; Spatial Distribution; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Staging; Technology; Testing; Tissues; Training; Tubular formation; Vascular Diseases; Work; base; career; design; diabetic; glomerulosclerosis; glycation; in vivo; inhibitor/antagonist; insight; mortality; new technology; novel; novel therapeutics; oxidative damage; pandemic disease; prevent; public health relevance; skills; small molecule; therapy development

DESCRIPTION (provided by applicant): In the following research training plan, I propose to apply imaging mass spectrometry to gain a detailed understanding of the molecular mechanisms underlying diabetic nephropathy (DN). DN is characterized by a number of pathological changes, including thickening of the glomerular and tubular basement membranes, mesangial expansion, glomerular sclerosis, arteriolar hyalinosis, tubular hypertrophy, and tubulointerstitial fibrosis. Although the morphological changes that occur in DN have been well defined, information about the molecular mechanisms governing this pathology is lacking. A number of biochemical events have been suggested to play a role in DN pathogenesis, including nonenzymatic glycooxidative damage of endogenous proteins. While the chemistries of glycooxidative reaction pathways are well understood, the specific reactions that occur in vivo and the roles that they play in DN are not well defined. We hypothesize that early glyco-oxidative modifications to extracellular proteins induced by hyperglycemia play a role in the development of DN. Because oxidative damage may occur via multiple pathways in DN, it is important to pursue novel analytical technologies that have the ability to monitor diverse biochemical changes in vivo. MALDI imaging mass spectrometry (IMS) is a new technology that allows for direct profiling of molecules in situ and is uniquely suited for analysis of a complex, multi-component disease like DN. Despite numerous studies of soluble proteins, peptides, and lipids, there are no reported IMS analyses of ECM proteins, to date. Therefore, the first goal of the proposed research will be to develop new protocols for IMS analysis of renal ECM proteins in tissue. Once established, these methods will be used to identify specific glycooxidative modifications that accompany progressive DN, in order to gain a better understanding of how glycation reactions contribute to the clinical features of renal failure in diabetes. IMS will allo for analysis of early glycooxidative modifications, before noticeable morphological changes appear. Finally, administration of pyridoxamine, a proven inhibitor of DN, to DN-prone mice will help to identify specific glycooxidative pathways that are connected to the pathology of DN, providing significant insight into the molecular mechanisms governing this disease. In addition to designing the research proposal, I have worked closely with my sponsor, Dr. Billy Hudson to develop a training plan for my professional development that will aid in my pursuit of a career as an independent researcher at an academic institution. I am confident that the research training plan that I have designed will aide in my development as a scientist and I am excited to apply the skills I will gain over the course of the fellowship towards a successful research career using mass spectrometry to investigate the molecular basis of renal and vascular diseases.

描述（由申请人提供）：在以下研究培训计划中，我建议采用成像质谱法以详细了解糖尿病性肾病（DN）的分子机制。 DN的特征是多种病理变化，包括肾小球和管状基底膜的增厚，肾小球膨胀，肾小球硬化症，小动脉透明质酸，管状肥大和管状纤维纤维纤维化。尽管DN中发生的形态学变化已得到很好的定义，但缺乏有关该病理学的分子机制的信息。已经建议许多生化事件在DN发病机理中发挥作用，包括内源性蛋白的非酶糖化损伤。虽然已经充分了解了糖氧化反应途径的化学途径，但体内发生的特定反应及其在DN中扮演的作用尚未得到很好的定义。 我们假设早期的糖氧化剂对高血糖诱导的细胞外蛋白质的改性在DN的发展中起作用。由于氧化损伤可能通过DN中的多种途径而发生，因此追求具有监测体内多种生化变化的新型分析技术很重要。 MALDI成像质谱法（IMS）是一项新技术，可直接对原位分子进行分析，并且非常适合分析复杂的多组分疾病（如DN）。 尽管对可溶性蛋白，肽和脂质进行了大量研究，但迄今为止，尚无IMS分析ECM蛋白的分析。因此，拟议的研究的第一个目标是开发组织中肾脏ECM蛋白IMS分析的新方案。建立后，这些方法将用于识别伴随进行性DN的特定糖氧化修饰，以便更好地了解糖化反应如何促进糖尿病肾衰竭的临床特征。 IMS将在出现明显的形态变化之前对早期糖氧化修饰进行分析。最后，对DN易发的小鼠的吡id胺（一种已验证的DN抑制剂）的给药将有助于鉴定与DN病理相关的特定糖氧化途径，从而对控制这种疾病的分子机制有很大的见解。 除了设计研究建议外，我还与赞助商Billy Hudson博士紧密合作，为我的专业发展制定了培训计划，这将有助于我从事学术机构的独立研究人员的职业。我有信心，我设计的研究培训计划将有助于我作为一名科学家的发展，我很高兴能在研究金的过程中运用我将获得的技能 质谱法研究了肾脏和血管疾病的分子基础。