Mitochondrial Therapy for Kidney Injury

项目摘要或摘要 预防和治疗急性肾损伤（阿基）仍然是一个重要的健康问题。 相当多的数据表明，免疫系统介导阿基，并且抗炎性免疫应答的发展是由免疫系统介导的。 治疗可以显著减轻组织损伤和功能丧失。然而，常见的副作用 抗炎治疗与缺乏临床数据相结合，支持免疫系统的参与。 系统在阿基发病机制中的作用，阻碍了抗炎选择的发展。线粒体， 阿基中的关键参与者具有作为主要能量来源（ATP）和作为细胞死亡的关键调节剂的双重作用。 肾缺血再灌注（IRI）诱导近端小管30-40%的线粒体断裂 (PT)细胞沿着线粒体生物合成调节因子PGC 1 β（过氧化物酶体增殖物激活受体γ- 共活化剂1 μ g）。全身静脉注射荧光染料标记的分离的健康线粒体信号是 早在注射后15分钟就在包括脾、肾、肝和肺的各种组织中发现。肾脏 分离的标记的线粒体主要被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， 免疫和肾脏驻留细胞反应，以防止损伤（治疗前）和诱导恢复（治疗后）。 可能涉及通过诱导PGC 1 α激活受体线粒体生物合成。的 转移的线粒体1）增强受体细胞的ATP产生和耗氧速率，2）定位 3）减少炎症反应（细胞死亡、细胞因子、炎性细胞浸润），和4） 诱导受体细胞PGC 1 β，肾反应的关键决定因素，以保护肾脏免受阿基。 因此，直接线粒体转移作为一种治疗干预，以修复，重编程和替换 线粒体、改善线粒体健康和恢复呼吸功能有益于预防和/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PGC 1 α和拯救来帮助替换受损的线粒体 细胞功能。我们将测试我们的假设与以下三个目标：目标1：为了测试假设， 用分离的健康线粒体治疗有助于维持细胞功能以阻断（预处理）阿基， 在小鼠阿基的两种模型（IRI和LPS诱导的脓毒症）中预防（治疗后）疾病进展。 目的2：验证健康线粒体摄取诱导线粒体代谢基因的假说 (PGC1α）增加线粒体数量和细胞内ATP，以防止阿基。目标3：测试 用分离的线粒体治疗有助于维持较大动物（猪）的细胞功能的假说 保护肾脏免受IRI的侵害。

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.