Mitochondrial-Targeted Therapeutics for Treatment of Spinal Cord Injury

线粒体靶向疗法治疗脊髓损伤

基本信息

批准号：
8447510
负责人：
Alexander George Rabchevsky
金额：
$ 31.35万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-06-15 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8447510
关键词：
Acetylcysteine Acute Amides Antioxidants Behavioral Bioenergetics Carnitine Cell Count Cells Collaborations Contusions Data Dose Enzymes Event Glutathione Healthcare Histopathology Homeostasis Impairment Injury Intervention Levocarnitine Acetyl Limb structure Maintenance Methylprednisolone Mitochondria Mitochondrial Proteins N-Acetylcysteinamide Neuroglia Neurons Outcome Measure Oxidative Phosphorylation Population Production Publishing Rattus Reactive Oxygen Species Recovery Recovery of Function Reporting Site Source Spinal cord injury Synapses Techniques Testing Therapeutic Tissues United States Universities Work base central nervous system injury combinatorial design dosage excitotoxicity improved kinematics mitochondrial dysfunction neuronal cell body neuroprotection novel novel therapeutic intervention oxidative damage prevent research study response therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Spinal cord injury (SCI) is a long-term health care problem in the United States, and with the exception of the modestly effective methylprednisolone, there is currently no neuroprotective intervention clinically available for treatment of acute SCI. Our published data and preliminary results demonstrate that oxidative damage to key mitochondrial enzymes and subsequent mitochondrial dysfunction is key to the neuropathological sequalae following SCI. This proposal focuses on directly targeting mitochondrial dysfunction as a novel therapeutic intervention for contusion SCI, the fundamental concept being that SCI-induced excitotoxicity increases mitochondrial Ca2+ cycling/overload and the production of reactive oxygen species (ROS), ultimately leading to mitochondrial dysfunction and glutathione (GSH) depletion. Our approach is two-pronged, aimed at reducing mitochondrial ROS production utilizing a novel, cell-permeant antioxidant and GSH precursor, NACA (the amide form of N-acetylcysteine), as well as an alternative biofuel substrate for energy production, acetyl-l-carnitine (ALC), following SCI. Our published and preliminary data signify that both NACA and ALC improve mitochondrial bioenergetics following contusion SCI in rats, and that prolonged NACA or ALC treatment increases tissue sparing following injury. The planned experiments are designed to test the novel hypothesis that reducing oxidative damage to key mitochondrial proteins will maintain mitochondrial bioenergetics, thus leading to increased neuroprotection and improved functional recovery following contusion SCI. Specifically we will: 1) Characterize oxidative damage to specific mitochondrial proteins involved in bioenergetics and test the hypothesis that NACA treatment ameliorates mitochondrial oxidative damage following SCI, 2) Test the hypothesis that a combinatorial treatment with NACA and ALC will act synergistically to preserve mitochondrial homeostasis following SCI, and 3) Test the hypothesis that a combinatorial treatment with NACA and ALC will increase tissue sparing and promote long-term functional recovery following SCI. Critically, this application is built around the utilization of several novel techniques we have developed for isolating synaptic (neuronal) and non-synaptic (soma and glia) mitochondria from the injured spinal cord, as well as an L1/L2 contusion SCI paradigm that demonstrates a significant correlation between neuroprotection and remarkable improvements in recovery of hind limb function. Collectively, the proposed experiments will pinpoint key mitochondrial events that could be potential novel targets for pharmacological interventions to more effectively treat SCI and, perhaps, other CNS injuries.

描述（申请人提供）：脊髓损伤（SCI）在美国是一个长期的医疗保健问题，除了效果不大的甲基强的松龙外，目前临床上还没有可用于治疗急性 SCI 的神经保护干预措施。我们发表的数据和初步结果表明，关键线粒体酶的氧化损伤和随后的线粒体功能障碍是 SCI 后神经病理学后遗症的关键。该提案的重点是直接针对线粒体功能障碍作为挫伤性 SCI 的一种新型治疗干预措施，基本概念是 SCI 诱导的兴奋性毒性增加线粒体 Ca2+ 循环/过载和活性氧 (ROS) 的产生，最终导致线粒体功能障碍和谷胱甘肽(谷胱甘肽)耗尽。我们的方法是双管齐下的，旨在利用新型细胞渗透性抗氧化剂和 GSH 前体 NACA（N-乙酰半胱氨酸的酰胺形式）以及用于能量生产的替代生物燃料底物乙酰-L 来减少线粒体 ROS 的产生。 -肉碱 (ALC)，遵循 SCI。我们发表的初步数据表明，NACA 和 ALC 均可改善大鼠挫伤性 SCI 后的线粒体生物能学，并且延长 NACA 或 ALC 治疗可增加损伤后的组织保护。计划中的实验旨在测试新的假设，即减少关键线粒体蛋白的氧化损伤将维持线粒体生物能，从而增强神经保护并改善挫伤性 SCI 后的功能恢复。具体来说，我们将：1) 表征生物能学中涉及的特定线粒体蛋白的氧化损伤，并检验 NACA 治疗可改善 SCI 后线粒体氧化损伤的假设，2) 检验 NACA 和 ALC 的组合治疗将协同作用以维持线粒体稳态的假设SCI 后，3) 检验 NACA 和 ALC 联合治疗将增加组织保留并促进 SCI 后长期功能恢复的假设。重要的是，该应用程序是围绕利用我们开发的几种新技术来从受伤的脊髓中分离突触（神经元）和非突触（体细胞和神经胶质细胞）线粒体以及 L1/L2 挫伤 SCI 范式而构建的神经保护与后肢功能恢复的显着改善之间存在显着相关性。总的来说，拟议的实验将查明关键的线粒体事件，这些事件可能成为药物干预的潜在新目标，以更有效地治疗 SCI 以及其他中枢神经系统损伤。