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: 9235872
• 负责人: ROSLYN B MANNON
• 金额: --
• 依托单位: BIRMINGHAM 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 2020-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9235872
• 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; 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; 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; Nephrotoxic; 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; exhaustion; improved; in vivo; inflammatory marker; injury and repair; insight; interstitial; kidney cell; kidney preservation; knock-down; mRNA Expression; macrophage; mitochondrial dysfunction; mouse model; nephrotoxicity; novel; novel therapeutic intervention; novel therapeutics; organ transplant rejection; paracrine; 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.

实体器官移植需要终身免疫抑制。特别是钙调神经磷酸酶 抑制剂（CNI）几乎用于所有移植受者，因为它们是有效的免疫抑制剂。在 肾移植时，使用 CNI 的排斥率非常低，每年约 7-10%。然而， 长期移植物存活率并没有随着其使用而显着改善，平均半衰期为 10 年 肾脏移植。晚期移植物损伤的一个关键因素是 CNI 肾毒性，其表现为肾功能下降 肾功能和间质纤维化、肾小管萎缩和小动脉透明变性的组织学特征。这 CNI引起肾损伤和衰竭的机制是多因素的，迄今为止，尚无具体的机制 减轻这种损伤的治疗策略。本次提交的目的是确定一种新颖且具有临床意义的药物 改善 CNI 相关肾损伤的适用策略。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因子，与 调节肾损伤。此外，AMPK 激活的丧失与器官炎症和纤维化有关。 我们假设 AMPK 激活可以改善肾脏疾病的一些不良代谢后果。 管状上皮细胞，作为推论，AMPK 激活可能是一种临床相关的干预措施，可减轻 长期 CNI 肾毒性。我们的研究将重点关注环孢素 A (CsA)，一种经典的 CNI 治疗药物。 我们将利用 CsA 损伤小鼠模型进行一系列补充性体外和体内研究 我们已经在实验室成功使用了。在体外，我们将重点关注肾小管上皮，即初级肾小管上皮。 体内损伤的目标。我们将研究 AMPK 激活以及敲低的影响 药物制剂和沉默 RNA。结果将包括生物能量学、线粒体的评估 功能、促炎标记物和 DAMP HMGB1，以及对巨噬细胞及其旁分泌的影响 差异化。在体内，我们将通过评估来测试 AMPK 激活对改善 CsA 肾损伤的影响 肾功能（血清肌酐）、肾脏内炎症介质的 mRNA 表达，以及 损伤和纤维化的生化、细胞和组织学评估。我们将评估先发制人 抑制以及确定 CsA 肾毒性后 AMPK 激活的影响。到 为了补充这些药理学研究，我们还将采用 AMPK α1 和 α2 缺陷的新用途 老鼠。交叉移植研究将使我们能够剖析 AMPK 的全身表达与肾脏表达的作用 受伤。我们相信，这种综合方法将为改善或改善提供关键的机制见解。 减轻 CNI 肾毒性。使用临床相关激活剂二甲双胍将提供机会 快速转化为肾脏和其他实体器官移植的人类接受者。