Mitochondrial injury interferes with endogenous renal repair in experimental renovascular disease

线粒体损伤干扰实验性肾血管疾病的内源性肾修复

• 批准号: 9805789
• 负责人: Alfonso Eirin
• 金额: $ 11.93万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9805789
• 关键词: Active Biological Transport; Acute Renal Failure with Renal Papillary Necrosis; Adopted; Adult; Affect; Animal Model; Apoptosis; Attenuated; Award; Cell physiology; Cells; Characteristics; Chronic; Chronic Kidney Failure; Data; Deterioration; Development; Diet; Disease; End stage renal failure; Energy Supply; Epithelial Cells; Family suidae; Fibrosis; Functional disorder; Future; Goals; Grant; Human; Hyperlipidemia; Impairment; Incubated; Individual; Injury; Insulin Resistance; Ischemia; K-Series Research Career Programs; Kidney; Kidney Diseases; Kidney Failure; Knowledge; Magnetic Resonance Imaging; Metabolic; Metabolic syndrome; Mitochondria; Modeling; Mus; National Institute of Diabetes and Digestive and Kidney Diseases; Natural regeneration; Obesity; Organelles; Oxidation-Reduction; Oxidative Stress; Patients; Peptides; Phenotype; Proliferating; Recovery; Renal Artery Stenosis; Renal function; Reperfusion Injury; Research Personnel; Secondary Hypertension; Structural defect; Structure; System; Techniques; Testing; Thinness; Time; Tubular formation; aging population; career; cell injury; effective intervention; efficacy study; improved; improved functioning; in vivo; innovation; kidney fibrosis; kidney repair; kidney vascular structure; mitochondrial dysfunction; mouse model; novel; novel strategies; paracrine; preservation; prevent; process repeatability; progenitor; programs; public health relevance; renal ischemia; repaired; skills; solute; tool

This application responds to PAR-18-103, “Small Grant Program for NIDDK K01/K08/K23 Recipients (R03)”, which provides NIDDK-supported K01, K08, and K23 recipients support at some point during the final two years of their K award, as they complete their transition to fully independent investigator status. Renovascular disease (RVD) remains an important cause of renal failure in the aging population. My K08 studies showed that RVD in swine induces mitochondrial injury in tubular cells, but whether this reflects the direct effect of ischemia and metabolic abnormalities on tubular cells or defective endogenous repair in the post-stenotic kidney is unknown. RVD presents repeated episodes of insults mimicking acute kidney injury (AKI) from which the kidney can normally recover. CD133+/CD24+ scattered tubular-like cells (STCs) represent a dedifferentiated phenotype that can be adopted by surviving tubular epithelial cells. STCs can proliferate and re-differentiate to replace lost neighboring cells. Our preliminary data suggest that RVD induces structural and functional abnormalities in the swine STC mitochondria, but whether RVD impairs the recovery potential of the kidney is unknown. Detecting STC dysfunction and elucidating the mechanisms responsible may facilitate development of adequate tools to preserve their reparative potency and kidney vitality. The hypothesis underlying this proposal is that RVD induces STC mitochondrial injury that impairs their integrity and function, blunting their overall capacity to repair the kidney. We will employ novel swine models of renal artery stenosis (RAS), metabolic syndrome (MetS), and MetS+RAS that closely mimic the ischemic and metabolic components of human RVD. We will study the efficacy of STCs in a murine model of ischemia reperfusion injury (IRI)-induced AKI. Delivery of swine STCs pretreated with mitochondria-targeted peptides (MTPs) will establish the contribution of mitochondrial dysfunction to STC impairment in RVD. Two specific aims will be pursued: Specific Aim 1 will test the hypothesis that RVD-induced STC mitochondrial injury affects their integrity and function. STCs will be collected from pigs after 16 weeks of Lean or MetS diet with or without RAS. Mitochondrial structure and function, as well as cellular injury and function will be assessed in primary cultures of STCs with and without pre-incubation with MTPs. Specific Aim 2 will test the hypothesis that RVD-induced mitochondrial injury in STCs blunts their capacity to repair in-vivo kidneys after AKI. AKI (IRI) will be induced in mice, and STCs (from Specific Aim 1) or vehicle will be intra-arterially injected 3 days after AKI. Renal function and fibrosis will be studied 2 weeks later. The proposed studies could greatly advance our understanding the vulnerability of this repair system and contribute towards development of strategies for improving the utility and efficacy of kidney repair in renal disease. This proposal is well aligned with the applicant’s career goals and will provide him with unique skills from theoretical and experimental knowledge to technical proficiency for his future independent career.

此应用程序响应PAR-18-103，“NIDDK K01/K08/K23获奖者小额助学金计划(R03)”， 在最后两个阶段的某个时间点提供NIDDK支持的K01、K08和K23接收方支持 随着他们完成向完全独立的调查员身份的过渡，他们获得了数年的K奖。 肾血管疾病(RVD)仍然是老年人口肾功能衰竭的重要原因。我的K08 研究表明，猪RVD可诱导肾小管细胞线粒体损伤，但这是否反映了 缺血和代谢异常对肾小管上皮细胞或内源性修复缺陷的直接影响 狭窄后的肾脏尚不清楚。RVD表现出反复发作的侮辱行为，模仿急性肾损伤 (AKI)肾脏可以正常恢复的。CD133+/CD24+散在小管样细胞 代表一种可被存活的肾小管上皮细胞采用的去分化表型。STC可以 增殖并重新分化，以取代丢失的相邻细胞。我们的初步数据表明，RVD导致 猪STC线粒体结构和功能异常，但RVD是否损害恢复 肾脏的潜力尚不清楚。检测STC功能障碍并阐明相关机制 可促进开发适当的工具，以保持其修复能力和肾脏活力。这个 支持这一建议的假设是，RVD导致STC线粒体损伤，从而损害其完整性 和功能，削弱了他们修复肾脏的整体能力。我们将采用新的猪肾脏模型。 动脉狭窄(RAS)、代谢综合征(METS)和METS+RAS，与缺血性和 人类RVD的代谢成分。我们将研究STCs在小鼠缺血模型中的疗效 再灌注损伤(IRI)诱导的AKI。线粒体靶向多肽处理的猪干细胞的传递 (MTP)将确定线粒体功能障碍在RVD患者STC损伤中的作用。两个具体的 将追求目标：特定目标1将检验RVD导致STC线粒体损伤的假设 影响它们的完整性和功能。精益日粮或蛋氨酸日粮16周后从猪身上收集STC，或 没有RAS。线粒体的结构和功能，以及细胞损伤和功能将在 含和不含MTPs预孵育的STCs原代培养。《特定目标2》将检验这一假设 RVD诱导的STCs线粒体损伤削弱了它们在AKI后修复体内肾脏的能力。Aki(IRI) 将在小鼠体内诱导，并在3天后动脉内注射STCs(来自特定目标1)或载体 阿琪。肾功能和纤维化将在2周后进行研究。拟议的研究将极大地推动我们的 了解这种修复系统的脆弱性，并为制定战略做出贡献 提高肾脏修复在肾脏疾病中的效用和疗效。这项建议与 申请者的职业目标，并将为他提供从理论和实验知识到 为他未来的独立职业生涯提供技术支持。