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，我建议应用成像质谱法来详细了解糖尿病肾病（DN）的分子机制。 DN以多种病理改变为特征，包括肾小球和肾小管基底膜增厚、系膜扩张、肾小球硬化、小动脉透明变性、肾小管肥大和肾小管间质纤维化。尽管 DN 中发生的形态学变化已得到明确定义，但仍缺乏有关控制这种病理学的分子机制的信息。许多生化事件被认为在 DN 发病机制中发挥作用，包括内源蛋白的非酶糖氧化损伤。虽然糖氧化反应途径的化学原理已被充分了解，但体内发生的具体反应及其在 DN 中发挥的作用尚不清楚。 我们假设高血糖诱导的细胞外蛋白的早期糖氧化修饰在 DN 的发展中发挥作用。由于 DN 中氧化损伤可能通过多种途径发生，因此寻求能够监测体内多种生化变化的新型分析技术非常重要。 MALDI 成像质谱 (IMS) 是一项新技术，可直接对分子进行原位分析，特别适合分析 DN 等复杂的多成分疾病。 尽管对可溶性蛋白质、肽和脂质进行了大量研究，但迄今为止还没有 ECM 蛋白质的 IMS 分析报道。因此，本研究的首要目标是开发用于组织中肾 ECM 蛋白 IMS 分析的新方案。一旦建立，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导致糖尿病肾衰竭的临床特征。 IMS 将允许在出现明显的形态变化之前分析早期糖氧化修饰。最后，对易患 DN 的小鼠施用吡哆胺（一种经过验证的 DN 抑制剂）将有助于识别与 DN 病理学相关的特定糖氧化途径，从而为控制该疾病的分子机制提供重要见解。 除了设计研究计划外，我还与我的资助者 Bil​​ly Hudson 博士密切合作，为我的专业发展制定培训计划，这将有助于我在学术机构成为一名独立研究员。我相信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，并且我很高兴能够运用我在奖学金过程中获得的技能，以实现成功的研究生涯 质谱法研究肾脏和血管疾病的分子基础。