Soluble (pro)renin receptor regulation of kidney fibrosis

可溶性肾素（原）受体对肾纤维化的调节

• 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
• 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

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.

摘要 据估计，美国有3700万人患有慢性肾病(CKD)。 慢性肾脏病进展涉及炎症和纤维化反应的激活，导致不可逆转 肾功能受损和丧失。(Pro)肾素受体(PRR)参与了 CKD的发病机制可以以全长形式存在，结合到细胞膜上或被切割 产生可溶性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得到了一定程度的研究，其病理生理 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 已在CKD患者中被描述，并与CKD的分期相关。我们最近 利用CRISPR-Cas9基因建立了一种新型的缺失sPRR的小鼠模型 PRR裂解位点的突变。初步分析显示，突变的sPRR小鼠 与对照组小鼠相比，肾脏损伤、炎症和纤维化程度降低，可能涉及 炎症信号和氧化磷酸化途径。以下具体目标将 收信人： 1.探讨sPRR在肾脏疾病中的病理生理作用。CKD将在 对照组和突变型sPRR小鼠用腺嘌呤饮食或单侧输尿管梗阻。肾 功能、肾小管损伤、炎症和纤维化将与 有针对性的比较转录学，以确定活跃的信号通路。 2.探讨sPRR调节肾损伤的细胞机制。SPRR 炎症信号通路和氧化磷酸化的调节 线粒体功能将在对照的原代近端小管细胞培养中进行检测 和有腺嘌呤或转化生长因子-存在的突变型sprr小鼠。将添加重组sPRR 对照和缺乏sPRR的突变细胞，以检查恢复sPRR水平是否逆转 肾脏保护作用。 3.研究sPRR的分子相互作用伙伴。SPRR如何调节细胞内 细胞信号将通过确定蛋白质-蛋白质相互作用的结构来进行研究。 导向亲和纯化、质谱学和酶催化邻近标记 在HEK293细胞中。质谱学和蛋白质组学分析将描绘sPRR蛋白 生理条件下和肾脏疾病中的相互作用和信号传递。 这项提案研究了一种新的肾脏损伤和纤维化的调节剂，并将描绘出 参与调节这些影响的机制。本文使用的综合方法将 明确sPRR在纤维化中的系统和分子效应，并可能有助于 慢性肾脏病的新治疗方法。