Mitochondrial Therapy for Kidney Injury

项目摘要或摘要 预防和治疗急性肾损伤(AKI)仍然是一个重大的健康问题。 大量数据表明，免疫系统介导了AKI和抗炎的发展 治疗可以显著减轻组织损伤和功能丧失。然而，共同的副作用 抗炎治疗结合缺乏临床数据，支持免疫参与 系统在AKI的发病机制中，有阻碍抗炎发展的选择。线粒体， AKI中的关键角色具有作为主要能量来源(ATP)和细胞死亡的关键调节因素的双重作用。 肾缺血再灌流(IRI)诱导近端小管线粒体碎裂 (PT)细胞以及线粒体生物发生调控因子Pgc1(PPARγ- 辅活化子1)。全身静脉注射荧光标记的分离健康线粒体信号 早在注射后15分钟，就在包括脾、肾、肝和肺在内的各种组织中发现了这种病毒。在肾脏中 分离的标记线粒体主要被PT摄取。我们的初步研究表明， 从健康来源(肌肉或肝脏)分离的健康线粒体的潜在治疗作用 免疫和肾脏常驻细胞反应，以防止损伤(前处理)和诱导恢复(后 治疗)，这可能涉及通过诱导pGc1α激活受体线粒体的生物发生。这个 转移的线粒体1)提高受体细胞的ATP产量和耗氧率，2)定位 对于肾脏，3)减少炎症反应(细胞死亡、细胞因子、炎性细胞浸润)，以及4) 诱导受体细胞Pgc1，肾脏反应的关键决定因素，以保护肾脏免受急性肾损伤。 因此，直接线粒体转移作为一种治疗干预手段来修复、重新编程和替换 线粒体，改善线粒体健康，恢复呼吸功能，有利于预防和/或 疾病的治疗。因此，我们假设用健康的线粒体治疗可以增强 受体细胞产生能量，通过诱导pGc1α和拯救来帮助修复受损的线粒体 细胞功能。我们将通过以下三个目标来检验我们的假设：目标1：检验以下假设 用分离的健康线粒体进行治疗有助于维持细胞功能，以阻断(预处理)AKI和 在两种AKI小鼠模型(IRI和脂多糖诱导的脓毒症)中预防(治疗后)疾病的进展。 目的2：验证摄取健康线粒体诱导线粒体代谢基因的假说 (前列环素α)，增加线粒体数量和细胞内三磷酸腺苷，以保护免受急性脑损伤。目标3：测试 假设用分离的线粒体治疗有助于维持较大动物(猪)的细胞功能 以保护肾脏免受IRI的影响。

Project Summary or Abstract Prevention and treatment of acute kidney injury (AKI) continues to remain a significant health problem. Considerable data suggest that the immune system mediates AKI, and development of anti-inflammatory treatments can significantly attenuate tissue injury and loss of function. However, the side effects of common anti-inflammatory therapies combined with the lack of clinical data, supporting the involvement of the immune system in AKI pathogenesis, have hindered the development of anti-inflammatory options. Mitochondria, critical players in AKI, have dual roles as a primary source of energy (ATP) and as key regulators of cell death. Renal ischemia followed by reperfusion (IRI) induces mitochondrial fragmentation in 30-40% of proximal tubule (PT) cells along with reduced expression of the mitochondrial biogenesis regulator, PGC1 (PPARγ- coactivator 1). Systemic intravenous injection of fluorochrome labeled isolated healthy mitochondria signal is found in various tissue including spleen, kidneys, liver and lungs as early as 15 mins after injection. In kidneys the isolated labeled mitochondria are predominantly taken up by PT. Our preliminary studies demonstrate a potential therapeutic role for healthy mitochondria isolated from a healthy source (muscle or liver) to alter immune and kidney resident cellular responses to prevent injury (pretreatment) and induce recovery (post- treatment) that may involve activation of recipient mitochondria biogenesis through induction of PGC1α. The transferred mitochondria 1) enhance ATP production and oxygen consumption rate of recipient cell, 2) localize to the kidneys, 3) reduce inflammatory responses (cell death, cytokines, inflammatory cell infiltration), and 4) induce recipient cell PGC1, a pivotal determinant of renal responses, to protect kidneys from AKI. Therefore, direct mitochondrial transfer as a therapeutic intervention to repair, reprogram and replace mitochondria, improve mitochondrial health and restore respiratory function is beneficial for prevention and/or treatment of disease. Accordingly, we hypothesize that treatment with healthy mitochondria enhances recipient cell energy production to help replace damaged mitochondria by inducing PGC1α and rescues cellular functions. We will test our hypothesis with the following three aims: Aim 1: To test the hypothesis that treatment with isolated healthy mitochondria helps maintain cellular functions to block (pretreatment) AKI and prevent (post-treatment) progression of disease in two models (IRI and LPS-induced sepsis) of AKI in mice. Aim 2: To test the hypothesis that uptake of healthy mitochondria induces the mitochondrial metabolism gene (PGC1α) to increase mitochondrial numbers and intracellular ATP to protect from AKI. Aim 3: To test the hypothesis that treatment with isolated mitochondria helps maintain cellular functions in larger animals (swine) to protect kidneys from IRI.