Investigation of non-HIV Collapsing Glomerulopathy

非 HIV 塌陷性肾小球病的调查

• 批准号: 9750079
• 负责人: Sumant Singh Chugh
• 金额: $ 55.68万
• 依托单位: RUSH UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-05 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9750079
• 关键词: ANGPTL4 gene; Acceleration; Adriamycin PFS; Age; Animal Model; Binding Sites; Biological; Biopsy; Blood capillaries; Cell Nucleus; Cell membrane; Cessation of life; Cities; Development; Disease; Down-Regulation; End stage renal failure; Endothelial Cells; Endothelium; Epithelial Cell Proliferation; Etiology; Evaluation; Focal Segmental Glomerulosclerosis; Glomerular Capillary; Goals; HIV Infections; Human; In Vitro; Inbreeding; Infection; Injections; Integrin beta3; Integrins; Investigation; Kidney Failure; Kidney Transplantation; Knock-out; Knockout Mice; Mass Spectrum Analysis; Mediating; Mexico; Modeling; Molecular; Molecular Profiling; Morphology; Mouse Strains; Mus; Nephrosis; Nuclear Translocation; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Plasma; Plasma Proteins; Plasmapheresis; Play; Predisposition; Proteins; Proteinuria; Proteomics; Rat Strains; Rattus; Recombinants; Recurrence; Renal glomerular disease; Rodent; Rodent Model; Role; Sampling; Serum; Sprague-Dawley Rats; Stains; Structure; Transgenes; Transgenic Organisms; Transplantation; Up-Regulation; WNT Signaling Pathway; WT1 gene; base; gel electrophoresis; glomerular endothelium; in vitro testing; in vivo; insight; kidney biopsy; migration; nano; nephrotoxicity; novel therapeutics; offspring; podocyte; promoter; slit diaphragm; synergism; transcription factor; two-dimensional

Abstract Collapsing glomerulopathy is a severe form of human glomerular disease that frequently results in rapid progression towards end stage kidney failure. Whereas this disease can be caused by HIV infection and other causes, this proposal will focus on a specific form of this disease caused by circulating proteins (hereon referred to as CG). Although currently classified as a form of focal and segmental glomerulosclerosis(FSGS), there are several molecular and morphological differences that set CG apart from other forms of FSGS. The applicants developed the first ever animal model of CG by injecting plasma from CG patients into a specific stain of inbred Sprague Dawley rats (IMC rats) over a decade ago. Further exploration of this model, and correlation of its molecular changes with human kidney biopsies resulted in the development of new mechanism based models that will be utilized in this proposal. The proposal is structured to explore two distinct morphological aspects of CG, epithelial cell proliferation and glomerular capillary loop collapse, at a molecular level and also explore cross talk between these two components of CG. Finally, a proteomic approach will be used to identify and create a short list of circulating proteins that cause this disease. De-identified human plasma and controls, and plasmapheresis samples from patients with recurrent CG after transplantation (referred to as CG plasma) will be used for these studies. The overall goal of the proposed studies is to develop new insight into the molecular pathogenesis of CG so as to be able to develop new therapies for this disease. In Specific Aim 1, molecular mechanisms of podocyte proliferation induced by ZHX2 and TERT will be explored using podocyte specific ZHX2 transgenic rats and TERT knockout rats in the IMC background injected with CG plasma. In Specific Aim 2, a new model of CG developed in β5 integrin knockout mice will be used to study the role of β3 integrin in the development of glomerular capillary loop collapse. Mice deficient in β3 integrin and podocyte specific β3 integrin knockout mice will be used to study the role of β5 integrin in CG. Rrm2b and β5 integrin knockout mice will be interbred to study pathway synergies. Finally, crosstalk studies between the podocyte and the endothelium during the pathogenesis of CG will be explored. In Specific Aim 3, CG and control plasma will be depleted of 14 common plasma proteins, their ability to induce CG changes confirmed, and the putative circulating proteins identified using a proteomics approach comprising of DIGE and nano-LC MS/MS. A short list of proteins will be tested in vitro and in vivo using the above animal models for their ability to induce CG.

抽象的 塌陷性肾小球病是人类肾小球疾病的一种严重形式，经常导致 快速进展至终末期肾衰竭。而这种疾病可能是由 HIV感染和其他原因，本提案将重点关注引起这种疾病的特定形式 通过循环蛋白质（本文称为 CG）。尽管目前被归类为一种形式 局灶性节段性肾小球硬化症（FSGS），有多种分子和形态学特征 CG 与其他形式的 FSGS 的区别。申请人开发出了有史以来第一个 通过将 CG 患者的血浆注射到近交系 Sprague 的特定染色中来建立 CG 动物模型 十多年前，道利大鼠（IMC 大鼠）。对该模型的进一步探索以及相关性 它与人类肾活检的分子变化导致了新的开发 本提案中将使用基于机制的模型。 该提案旨在探索 CG 的两个不同形态学方面：上皮细胞 增殖和肾小球毛细血管袢塌陷，在分子水平上，并探索交叉 CG 的这两个组成部分之间的对话。最后，将使用蛋白质组学方法来鉴定 并创建一个导致这种疾病的循环蛋白的简短列表。身份不明的人类 血浆和对照，以及来自复发性 CG 患者的血浆去除样本 这些研究将使用移植（称为CG血浆）。 拟议研究的总体目标是开发对分子生物学的新见解 CG的发病机制，以便能够开发针对该疾病的新疗法。 具体目标1，ZHX2和TERT诱导足细胞增殖的分子机制 将使用足细胞特异性 ZHX2 转基因大鼠和 TERT 敲除大鼠在 IMC 背景注入 CG 血浆。 在Specific Aim 2中，在β5整合素敲除小鼠中开发的新CG模型将用于 研究β3整合素在肾小球毛细血管袢塌陷发展中的作用。小鼠 缺乏β3整合素和足细胞特异性β3整合素敲除小鼠将用于研究 β5整合素在CG中的作用。 Rrm2b和β5整合素敲除小鼠将进行杂交研究 途径协同作用。最后，足细胞和内皮细胞之间的串扰研究 将探讨CG的发病机制。 在特定目标 3 中，CG 和对照血浆将去除 14 种常见血浆蛋白，它们的 诱导 CG 变化的能力得到证实，并且使用 蛋白质组学方法包括 DIGE 和 nano-LC MS/MS。蛋白质的简短列表将是 使用上述动物模型在体外和体内测试其诱导 CG 的能力。