AMP Kinase Activation in Calcineurin Inhibitor Nephrotoxicity

钙调神经磷酸酶抑制剂肾毒性中的 AMP 激酶激活

• 批准号: 10187188
• 负责人: ROSLYN B MANNON
• 金额: --
• 依托单位: OMAHA VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-10-01 至 2021-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10187188
• 关键词: 5' -AMP-activated protein kinase; Acute Renal Failure with Renal Papillary Necrosis; Adverse effects; Affect; Allografting; Apoptosis; Atrophic; Autophagocytosis; Biochemical; Bioenergetics; Biogenesis; Bone Marrow; Calcineurin inhibitor; Caring; Cellular Metabolic Process; Chronic; Clinical; Clinical Management; Complement; Coupled; Creatinine; Cyclic AMP-Dependent Protein Kinases; Cyclosporine; Data; Diabetic Nephropathy; Dose; Duct (organ) structure; Epithelial; Epithelial Cells; Epithelium; Exposure to; Failure; Fibrosis; Functional disorder; Goals; Graft Survival; HMGB1 gene; Half-Life; Health; Heart; Histologic; Homeostasis; Human; Immune; Immunosuppression; Immunosuppressive Agents; In Vitro; Individual; Inflammation; Inflammation Mediators; Inflammatory; Injury; Injury to Kidney; Intervention; Kidney; Kidney Diseases; Kidney Failure; Kidney Transplantation; Laboratories; Laboratory mice; Lead; Link; Liver; Lung; Mediating; Mediator of activation protein; Medical center; Metabolic; Metformin; Mitochondria; Mus; NADPH Oxidase; Organ; Organ Transplantation; Organ failure; Outcome; Oxidation-Reduction; Oxidative Stress; Pancreas; Paracrine Communication; Pathway interactions; Patient-Focused Outcomes; Pharmaceutical Preparations; Pharmacology; Pharmacology Study; Polycystic Kidney Diseases; Population; Production; Property; Protein Kinase; RNA Interference; Reactive Oxygen Species; Renal function; Role; Series; Serum; Solid; Testing; Therapeutic; Therapeutic Agents; Therapeutic immunosuppression; Time; Tissues; Toxic effect; Translations; Transplant Recipients; Transplantation; Tubular formation; United States; Veterans; Whole Organism; adenylate kinase; cell injury; clinical application; clinically relevant; effective therapy; epithelial injury; exhaustion; improved; in vivo; inflammatory marker; injury and repair; insight; interstitial; kidney cell; kidney dysfunction; kidney preservation; knock-down; mRNA Expression; macrophage; mitochondrial dysfunction; mouse model; nephrotoxicity; novel; novel therapeutic intervention; novel therapeutics; organ transplant rejection; paracrine; preservation; prevent; programs; success

Solid organ transplantation necessitates the use of lifelong immunosuppression. In particular, the calcineurin inhibitors (CNIs) are utilized in nearly all transplant recipients as they are potent immunosuppressants. In kidney transplantation, the use of CNIs has led to the very low rejection rates of ~7-10% per year. However, long term graft survival has not improved substantially with their utilization, with a mean half-life of 10 years for kidney transplants. A key contributor to late graft injury is CNI-nephrotoxicity which is manifested by declining kidney function and the histological features of interstitial fibrosis, tubular atrophy, and arteriolar hyalinosis. The mechanism by which CNI induce kidney injury and failure is multifactorial, and to date, there are no specific therapeutic strategies to mitigate this injury. The goal of this submission is to identify a novel and clinically applicable strategy to ameliorate CNI-associated kidney injury. Recently, AMP-Activated Protein Kinase (AMPK), a key regulator of cell metabolism, autophagy, and mitochondrial biogenesis, has been linked to modulating kidney injury. Further, loss of AMPK activation is associated with organ inflammation and fibrosis. We hypothesize that AMPK activation could ameliorate some of the adverse metabolic consequences in renal tubular epithelium and as a corollary, AMPK activation could be a clinically relevant intervention to mitigate long term CNI nephrotoxicity. Our study will focus on cyclosporine A (CsA), a classic CNI therapeutic agent. We will utilize a complementary series of studies, both in vitro and in vivo, using a mouse model of CsA injury that we have used successfully in our laboratory. In vitro, we will focus on renal tubular epithelium, the primary target of in vivo injury. We will study both the impact of AMPK activation as well as knock down using pharmacologic agents and silencing RNAs. Outcomes will include assessments of bioenergetics, mitochondrial function, pro-inflammatory markers and the DAMP HMGB1, and paracrine impact on macrophages and their differentiation. In vivo, we will test the impact of AMPK activation on ameliorating CsA renal injury by assessing kidney function (serum creatinine), mRNA expression of inflammatory mediators within the kidney, and biochemical, cellular and histologic assessments of injury and fibrosis. We will assess both pre-emptive inhibition as well as determine the impact of AMPK activation after established CsA nephrotoxicity. To complement these pharmacologic studies, we will also employ the novel use of AMPK 1 and 2 deficient mice. Cross-transplant studies will allow us to dissect the role of systemic versus renal expression of AMPK in injury. We believe that this comprehensive approach will provide key mechanistic insights into ameliorating or mitigating CNI nephrotoxicity. The use of a clinically relevant activator, metformin, will provide an opportunity for rapid translation into our human recipients of kidneys and other solid organ transplants.

固体器官移植需要使用终身免疫抑制。特别是钙调蛋白 抑制剂（CNIS）几乎在所有移植受者中都使用，因为它们是潜在的免疫抑制剂。在 肾脏移植，CNIS的使用导致每年的排斥率非常低约7-10％。然而， 长期移植生存的利用情况并未大大提高，平均半衰期为10年 肾脏移植。引起疗法晚期损伤的关键因素是CNI-氯毒性，通过下降来表现 肾功能和间质纤维化，管状萎缩和Arterniolar Hyalinisois的组织学特征。 CNI影响肾脏损伤和失败的机制是多因素的，迄今为止，没有特定的 减轻这种伤害的治疗策略。此提交的目的是确定小说和诊所 适用于改善CNI相关肾脏损伤的策略。最近，AMP激活的蛋白激酶 （AMPK）是细胞代谢，自噬和线粒体生物发生的关键调节剂，已与 调节肾脏损伤。此外，AMPK激活的丧失与器官感染和纤维化有关。 我们假设AMPK激活可以改善肾脏中的某些不良代谢后果 肾小管上皮和推论，AMPK激活可能是临床上相关的干预措施，以减轻 长期CNI肾毒性。我们的研究将集中于经典的CNI治疗剂Cyclosporine A（CSA）。 我们将使用CSA损伤的小鼠模型在体外和体内使用一系列完整的研究 我们在实验室成功使用了。在体外，我们将专注于肾小管上皮，主要 体内损伤的靶标。我们将研究AMPK激活的影响以及使用 药理学剂和沉默的RNA。结果将包括对生物能学，线粒体的评估 功能，促炎标记和潮湿HMGB1以及旁分泌对巨噬细胞的影响及其 分化。在体内，我们将通过评估来测试AMPK激活对改善CSA肾脏损伤的影响 肾功能（血清肌酐），肾脏中炎症介质的mRNA表达和 损伤和纤维化的生化，细胞和组织学评估。我们将评估两个先发制人 抑制作用并确定建立CSA肾毒性后AMPK激活的影响。到 补充这些药理学研究，我们还将采用AMPK1和2的新颖使用 老鼠。跨移植研究将使我们能够剖析AMPK在 受伤。我们认为，这种全面的方法将为改善或 减轻CNI肾毒性。使用临床相关激活剂二甲双胍将提供机会 快速转化为我们的人类的孩子的接受者和其他固体器官移植。