Soluble (pro)renin receptor regulation of kidney fibrosis

可溶性肾素（原）受体对肾纤维化的调节

基本信息

批准号：
10745143
负责人：
NIRUPAMA RAMKUMAR
金额：
$ 45.16万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10745143
关键词：
Address Adenine Adult Affect Affinity Chromatography Angiotensin II Attenuated Binding Biochemical Blood Pressure CRISPR/Cas technology Cell Culture Techniques Cell membrane Cell secretion Cell surface Cells Chronic Disease Chronic Kidney Failure Development Diet Disease Disease Progression Enzymes Etiology Fibrosis Future Gene Expression Gene Expression Profiling Genetic Transcription Hypertension In Vitro Inflammation Inflammatory Inflammatory Response Infusion procedures Injury Injury to Kidney Interleukin-6 Kidney Kidney Diseases Label Length Mass Spectrum Analysis Mediating Mitochondria Molecular Morbidity - disease rate Mus Mutagenesis Oxidative Phosphorylation Pathogenesis Pathway interactions Patients Persons Physiological Plasma Population Proteins Proteomics Receptor, Angiotensin, Type 1 Recombinants Regulation Renal function Renin Renin-Angiotensin-Aldosterone System Role Signal Pathway Signal Transduction Site Structure System Techniques Time Tubular formation United States Ureteral obstruction Vascular Endothelial Cell autocrine cardiovascular risk factor comparative comparison control hypertensive in vivo inflammatory modulation innovation kidney fibrosis mortality mouse model mutant nano-string nectin novel novel therapeutic intervention organ injury paracrine receptor response transcriptome sequencing transcriptomics

项目摘要

ABSTRACT Chronic kidney disease (CKD) affects an estimated 37 million people in the United States. CKD progression involves activation of inflammatory and fibrotic responses leading to irreversible damage and loss of kidney function. The (pro)renin receptor (PRR) is implicated in the pathogenesis of CKD and can exist as the full length form, bound to cell membrane or be cleaved to generate a soluble PRR (sPRR) and M8.9 fragments. Although the function of the full-length PRR both at a molecular and system level has been studied to some extent, the pathophysiologic role of sPRR in CKD is unknown. This is especially important, since elevated plasma sPRR levels have been described in patients with CKD and correlates with the stage of CKD. We recently developed a novel mouse model with absence of sPRR using CRISPR-Cas9 directed mutagenesis of the PRR cleavage site. Preliminary analyses show mutant sPRR mice have reduced renal injury, inflammation and fibrosis compared to control mice and may involve inflammatory signaling and oxidative phosphorylation pathways. The following specific aims will be addressed: 1. Investigate the pathophysiological role of sPRR in kidney disease. CKD will be induced in control and mutant sPRR mice using adenine diet or unilateral ureteral obstruction. Renal function, tubular injury, inflammation and fibrosis will be examined in conjunction with targeted comparative transcriptomics to identify active signaling pathways. 2. Investigate the cellular mechanisms by which sPRR modulates kidney injury. sPRR regulation of inflammatory signaling pathways and oxidative phosphorylation and mitochondrial function will be examined in primary proximal tubule cell culture from control and mutant sPRR mice in presence of adenine or TGF-. Recombinant sPRR will be added to control and mutant cells lacking sPRR to examine if restoring sPRR levels reverses the renoprotective effects. 3. Investigate the molecular interaction partners of sPRR. How sPRR mediates intracellular cell signaling will be examine by identifying protein-protein interactors through structure- guided affinity purification and mass-spectrometry and enzyme catalyzed proximity labeling in HEK293 cells. Mass spectrometry and proteomics analyses will delineate sPRR protein interaction and signaling under physiological conditions and in kidney disease. This proposal examines a novel modulator of kidney injury and fibrosis and will delineate the mechanisms involved in mediating these effects. The integrative approach used herein will identify systemic and molecular effects of sPRR in fibrosis and may help in the development of a new therapeutic approach for CKD.

抽象的据估计，美国有 3700 万人患有慢性肾病 (CKD)。 CKD 进展涉及炎症和纤维化反应的激活，导致不可逆的肾素（原）受体（PRR）的损伤和丧失与肾功能有关。 CKD 的发病机制，可以以全长形式存在，与细胞膜结合或被切割生成可溶性 PRR (sPRR) 和 M8.9 片段的全长功能。 PRR在分子和系统水平上都已得到一定程度的研究，病理生理学 sPRR 在 CKD 中的作用尚不清楚，这一点尤其重要，因为血浆 sPRR 水平升高。我们最近在 CKD 患者中进行了描述，并且与 CKD 分期相关。使用 CRISPR-Cas9 开发了一种缺乏 sPRR 的新型小鼠模型 PRR 裂解位点的诱变初步分析显示突变的 sPRR 小鼠具有 PRR 裂解位点的突变。与对照小鼠相比，肾损伤、炎症和纤维化减少，并且可能涉及以下具体目标将是炎症信号传导和氧化磷酸化途径。予以解决： 1. 研究 sPRR 在 CKD 诱发中的病理生理学作用。使用腺嘌呤饮食或单侧输尿管梗阻的对照和突变 sPRR 小鼠。功能、肾小管损伤、炎症和纤维化将与比较靶向转录组学以确定活性信号通路。 2. 研究 sPRR 调节肾损伤的细胞机制。炎症信号通路和氧化磷酸化的调节将在原代近端小管细胞培养物中检查对照的线粒体功能将添加存在腺嘌呤或 TGF-β 的突变 sPRR 小鼠。对照和缺乏 sPRR 的突变细胞来检查恢复 sPRR 水平是否会逆转肾脏保护作用。 3. 研究 sPRR 的分子相互作用伙伴如何介导细胞内。将通过结构识别蛋白质-蛋白质相互作用物来检查细胞信号传导引导亲和纯化和质谱分析以及酶催化邻近标记 HEK293 细胞中的质谱和蛋白质组学分析将描述 sPRR 蛋白。生理条件下和肾脏疾病中的相互作用和信号传导。该提案研究了一种新型的肾损伤和纤维化调节剂，并将描述本文使用的综合方法将涉及调节这些效应的机制。确定 sPRR 在纤维化中的系统和分子效应，并可能有助于开发 CKD 的新治疗方法。