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.

抽象的慢性肾脏疾病（CKD）在美国影响约3700万人。 CKD进展涉及激活炎症和纤维化反应，导致不可逆肾功能的损害和丧失。（Pro）肾素受体（PRR）与 CKD的发病机理可以作为全长形式存在，与细胞膜结合或裂解生成固体PRR（SPRR）和M8.9片段。虽然全长的功能 PRR在分子水平和系统水平上都在某种程度上进行了研究，即病理生理 SPRR在CKD中的作用尚不清楚。这一点尤其重要，因为升高的血浆SPRR水平已经在CKD患者中描述了，并且与CKD的阶段相关。我们最近开发了一种新型的鼠标模型，没有使用CRISPR-CAS9的SPRR的新型模型 PRR裂解位点的诱变。初步分析表明，突变体SPRR小鼠具有与对照小鼠相比，肾脏损伤，感染和纤维化减少，可能涉及炎症信号传导和氧化磷酸化途径。以下具体目标将被解决： 1。研究SPRR在肾脏疾病中的病理生理作用。 CKD将被诱导使用腺嘌呤饮食或单侧输尿管反对的对照和突变体SPRR小鼠。肾脏功能，管状损伤，感染和纤维化将与靶向比较转录组学识别有效信号通路。 2。研究SPRR调节肾脏损伤的细胞机制。 sprr 调节炎症信号通路和氧化磷酸化和线粒体功能将在对照的原代近端小管细胞培养中检查在存在腺嘌呤或TGF-的情况下，突变的SPRR小鼠。将添加重组SPRR 控制和缺乏SPRR的突变细胞来检查恢复SPRR水平是否反转肾脏保护作用。 3。研究SPRR的分子相互作用伙伴。 SPRR如何介导细胞内通过结构 - 引导亲和力纯化，质谱法和酶催化接近标记在HEK293细胞中。质谱和蛋白质组学分析将描述SPRR蛋白在身体状况和肾脏疾病中的相互作用和信号传导。该提案研究了一个新型的肾脏损伤和纤维化调节剂，并将描绘介导这些作用的机制。这里使用的集成方法将确定SPRR在纤维化中的全身和分子效应，可能有助于发展 CKD的新治疗方法。