Proteinuria causes progressive kidney disease by impairing autophagy

蛋白尿通过损害自噬导致进行性肾脏疾病

基本信息

批准号：
8463517
负责人：
Andrea Havasi
金额：
$ 14.93万
依托单位：
BOSTON MEDICAL CENTER
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：
2011-07-15 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8463517
关键词：
Affect Albumin Receptors Albumins American Amino Acids Apoptosis Area Atrophic Autophagocytosis Award Biological Cell Death Cell physiology Cells Chronic Kidney Failure Clinical Complement Cytochromes Development Diabetic mouse Disease Progression Down-Regulation End stage renal failure Endocytosis Epithelial Epithelial Cells Event Extravasation Fibrosis Foundations Functional disorder Goals Hypertension Kidney Kidney Diseases Lead Link Mentors Mitochondria Modeling Molecular Mus Organ Organ failure Organelles Permeability Play Process Production Productivity Proteins Proteinuria Public Health Reactive Oxygen Species Research Research Support Role Signal Transduction Sirolimus Techniques Testing Therapeutic Toxic effect Tubular formation United States National Institutes of Health Up-Regulation base career cell injury cytochrome c experience extracellular human AMID protein human FRAP1 protein improved in vivo inhibition of autophagy inhibitor/antagonist interstitial macromolecule mitochondrial dysfunction mitochondrial membrane mouse model novel novel therapeutic intervention novel therapeutics oxidative damage pressure prevent pro-apoptotic protein receptor-mediated signaling research study response sensor tool

项目摘要

DESCRIPTION (provided by applicant): Proteinuria is associated with progressive chronic kidney disease. It is well known that exposure of proximal tubular epithelial (PTEC) cells to large amounts of albumin leads to the development of tubular atrophy and fibrosis. However, the possible pathogenic role of albumin in this process has not been fully elucidated. Development of new therapeutic tools to prevent or slow the progression of proteinuric chronic kidney disease requires clear understanding of the effect of proteinuria on tubular cell function. We propose that exposure of PTEC to albumin inhibits autophagy, a critical cellular function responsible for turnover of cellular macromolecules and organelles, including dysfunctional mitochondria. Our hypothesis is that albumin impairs autophagy by one of the following mechanisms: (a) stimulating albumin receptor-mediated signaling events; (b) up-regulating mTOR, a potent autophagy inhibitor; or (c) causing lysosomal dysfunction or inhibition of autophagosomal-lysosomal fusion which, in turn, inhibits autophagy. We hypothesize that experimental inhibition of autophagy by albumin overload or by knockdown of crucial autophagic proteins will result in accumulation of dysfunctional mitochondria leading to increased production of reactive oxygen species. The resulting oxidative damage increases the permeability and depolarization of the mitochondrial membrane and facilitates the leakage of intramitochondrial components such as cytochrome c and AIF, pro-apoptotic proteins that promote cell death. In in vivo experiments, proteinuria will be correlated with autophagy, ROS production, PTEC apoptosis, tubular atrophy, fibrosis and organ function. We will use Ins2Akita/+, a diabetic mouse model that develops proteinuria, interstitial fibrosis, tubular dysfunction and atrophy. We will also test if pharmacologic up-regulation of autophagy ameliorates proteinuria-induced changes in the proximal tubule. By investigating the possible signal transduction cascades that impair autophagy, this proposal will improve our understanding of the pathogenic role of albumin in proteinuric states, an established cause of progressive chronic kidney disease, and will investigate the novel role of autophagy stimulators in ameliorating progressive organ failure. Mastering the new principles and techniques presented in this proposal is essential for my scientific development and future research productivity. They will complement my previous experiences in the lab, permit me to lay down the foundation of a unique area of research and help me make a significant contribution to the depth and breadth of research in the field of proteinuric kidney diseases. I believe that this NIH mentored award will promote my scientific maturity and allow me to transition to a level that will make me competitive for independent research support.

描述（由申请人提供）：蛋白尿与进行性慢性肾病有关。众所周知，近端肾小管上皮（PTEC）细胞暴露于大量白蛋白会导致肾小管萎缩和纤维化。然而，白蛋白在此过程中可能的致病作用尚未完全阐明。开发新的治疗工具来预防或减缓蛋白尿慢性肾病的进展需要清楚地了解蛋白尿对肾小管细胞功能的影响。我们认为，PTEC 暴露于白蛋白会抑制自噬，这是一种负责细胞大分子和细胞器（包括功能失调的线粒体）更新的关键细胞功能。我们的假设是白蛋白通过以下机制之一损害自噬：（a）刺激白蛋白受体介导的信号事件； (b) 上调 mTOR，一种有效的自噬抑制剂； (c)引起溶酶体功能障碍或抑制自噬体-溶酶体融合，进而抑制自噬。我们假设通过白蛋白超载或敲低关键自噬蛋白来实验性抑制自噬将导致功能失调的线粒体积累，从而导致活性氧的产生增加。由此产生的氧化损伤增加了线粒体膜的通透性和去极化，并促进线粒体内成分（如细胞色素 c 和 AIF）的渗漏，这些成分是促进细胞死亡的促凋亡蛋白。在体内实验中，蛋白尿将与自噬、ROS产生、PTEC凋亡、肾小管萎缩、纤维化和器官功能相关。我们将使用 Ins2Akita/+，这是一种糖尿病小鼠模型，会出现蛋白尿、间质纤维化、肾小管功能障碍和萎缩。我们还将测试自噬的药理上调是否可以改善蛋白尿引起的近端小管变化。通过研究可能损害自噬的信号转导级联，该提案将提高我们对白蛋白在蛋白尿状态（进行性慢性肾病的既定原因）中致病作用的理解，并将研究自噬刺激剂在改善进行性器官衰竭中的新作用。掌握本提案中提出的新原理和技术对于我的科学发展和未来的研究生产力至关重要。它们将补充我以前在实验室的经验，使我能够为独特的研究领域奠定基础，并帮助我为蛋白尿肾病领域研究的深度和广度做出重大贡献。我相信，这个 NIH 指导的奖项将促进我的科学成熟度，并使我能够过渡到使我在独立研究支持方面具有竞争力的水平。