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.

摘要 塌陷性肾小球病是一种严重的人类肾小球疾病， 迅速发展为终末期肾衰竭而这种疾病可能是由 艾滋病病毒感染等原因，本建议将重点放在这种疾病的一种特定形式引起的 通过循环蛋白质（在此称为CG）。虽然目前被列为一种形式的 局灶节段性肾小球硬化症（FSGS），有几个分子和形态学 这些差异将CG与其他形式的FSGS区分开来。申请人开发了有史以来第一个 通过将来自CG患者的血浆注射到近交系Sprague的特定染色物中的CG动物模型 道利大鼠（IMC大鼠）。对该模型的进一步探讨， 它与人类肾脏活检的分子变化导致了新的 将在本提案中使用的基于机制的模型。 该建议的结构，以探讨两个不同的形态学方面的CG，上皮细胞 增殖和肾小球毛细血管袢塌陷，在分子水平上也探讨交叉 这是CG的两个组成部分之间的对话。最后，蛋白质组学方法将用于识别 并创建一个简短的列表，循环蛋白质，导致这种疾病。去识别人类 血浆和对照，以及来自复发性CG患者的血浆置换样品， 这些研究将使用移植物（称为CG血浆）。 拟议研究的总体目标是对分子生物学的新认识， CG的发病机制，以便能够开发这种疾病的新疗法。 在特异性目的1中，ZHX 2和TERT诱导足细胞增殖的分子机制 将使用足细胞特异性ZHX 2转基因大鼠和TERT敲除大鼠在 用CG等离子体注入IMC背景。 在具体目标2中，将使用在β5整联蛋白敲除小鼠中开发的CG新模型， 探讨β3整合素在肾小球毛细血管袢塌陷中的作用。小鼠 β3整合素缺陷和足细胞特异性β3整合素敲除小鼠将用于研究 β5整合素在CG中的作用。将Rrm 2b和β5整联蛋白敲除小鼠杂交以研究 路径协同作用。最后，足细胞和内皮细胞之间的串扰研究 探讨CG的发病机制。 在特定目标3中，CG和对照血浆将耗尽14种常见的血浆蛋白，其 诱导CG变化的能力得到证实，并使用免疫组织化学方法鉴定了推定的循环蛋白质。 蛋白质组学方法包括DIGE和纳米LC MS/MS。 使用上述动物模型在体外和体内测试它们诱导CG的能力。