Investigation of non-HIV Collapsing Glomerulopathy

摘要 塌陷性肾小球病变是一种严重的人类肾小球疾病，经常导致 迅速发展为终末期肾功能衰竭。鉴于这种疾病可由以下因素引起 HIV感染等原因，这项建议将重点针对这种疾病引起的一种特定形式 通过循环蛋白(以下称为CG)。尽管目前被归类为 局灶性节段性肾小球硬化(FSGS)，有几种分子和形态 区别，使CG有别于其他形式的FSGS。申请者开发出了有史以来第一个 近交系特殊染色注射CG患者血浆建立CG动物模型 道利大鼠(IMC大鼠)十多年前。对这一模型的进一步探索，以及与 它与人肾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。 该提案旨在探索CG的两个不同的形态方面，即上皮细胞 增殖与肾小球毛细血管环塌陷，在分子水平上也有交叉探讨 在CG的这两个组成部分之间进行交谈。最后，将使用蛋白质组学方法来鉴定 并创建一份导致这种疾病的循环蛋白的简短清单。未确认身份的人 血浆和对照组，以及血浆置换治疗后复发的CG患者的样本 移植(称为CG血浆)将用于这些研究。 拟议研究的总体目标是发展对分子的新见解。 探讨CG的发病机制，以期为该病开发新的治疗方法。 目的1.ZHX2和TERT诱导足细胞增殖的分子机制 将利用足细胞特异性ZHX2转基因大鼠和TERT基因敲除大鼠在 IMC背景注射CG血浆。 在特定目标2中，在β5整合素基因敲除小鼠中建立的新的CG模型将用于 探讨β-3整合素在肾小球毛细血管环塌陷中的作用。老鼠 β3整合素缺陷和足细胞特异性β3整合素基因敲除小鼠将被用于研究 β-5整合素在CG中的作用Rrm2b和β5整合素基因敲除小鼠将进行杂交研究 途径协同效应。最后，足细胞和内皮细胞之间的串扰研究 我们将对CG的发病机制进行探讨。 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，他们的 证实了诱导CG变化的能力，并使用A 蛋白质组学方法包括DGE和纳米LC MS/MS。 用上述动物模型进行体外和体内实验，检测其诱导CG的能力。

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.