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.

该申请对Par-18-103的响应，“ NIDDK K01/K08/K23收件人（R03）的小型赠款计划”， 在最后两点的某个时候，它提供了NIDDK支持的K01，K08和K23接收者的支持 他们的K奖颁发了多年，因为他们完成了向完全独立的调查员地位的过渡。 肾血管疾病（RVD）仍然是老龄化人群肾衰竭的重要原因。我的K08 研究表明，猪的RVD诱导管状细胞中的线粒体损伤，但这是否反映了 缺血和代谢异常对管状细胞或内源性修复的直接作用 静脉后肾脏未知。 RVD呈现出重复的侮辱情节，模仿急性肾脏受伤 （AKI）肾脏通常可以恢复。 CD133+/CD24+散射的管状细胞（STC） 代表可通过生存管状上皮细胞采用的去分化表型。 STC可以 增殖并重新分化以替代丢失的相邻细胞。我们的初步数据表明RVD诱导 猪STC线粒体中的结构和功能异常，但是RVD是否会损害恢复 肾脏的潜力是未知的。检测STC功能障碍并阐明负责的机制 可能有助于开发足够的工具来保留其修复效力和肾脏活力。这 该提案的基础假设是RVD诱导STC线粒体损伤，损害其完整性 和功能，使他们的整体修复肾脏能力钝化。我们将采用肾脏的新型猪模型 动脉狭窄（RAS），代谢综合征（MetS）和Mets+Ras，它们紧密模仿缺血性和 人RVD的代谢成分。我们将在缺血模型中研究STC的效率 再灌注损伤（IRI）引起的AKI。用线粒体靶向肽预处理的猪STC的递送 （MTP）将建立线粒体功能障碍对RVD中STC损伤的贡献。两个具体 将追求目标：特定目标1将检验RVD诱导的STC线粒体损伤的假设 影响他们的完整性和功能。在使用或大都会饮食16周后，将从猪中收集STC 没有拉斯。线粒体结构和功能以及细胞损伤和功能将在 具有和没有MTP预先孵育的STC的一级培养。具体目标2将检验假设 RVD引起的STC中的线粒体损伤钝化了AKI后修复体内肾脏的能力。 aki（iri） 将在小鼠中诱导，而STC（来自特定的目标1）或车辆将在三天后进行动力内注射 aki。两周后将研究肾功能和纤维化。拟议的研究可以极大地推动我们的 了解该维修系统的脆弱性，并有助于制定战略 改善肾脏疾病中肾脏修复的效用和效率。该建议与 申请人的职业目标，并将为他提供从理论和实验知识到 他未来独立职业的技术水平。