Pharmacologic treatment of mitochondrial complex I dysfunction in C. elegans

描述（由申请人提供）：线粒体呼吸链（RC）中复合物I的功能障碍是一组非常常见的多系统疾病的基础，这些疾病困扰着所有年龄和种族。相对不能客观地评估介导广泛变化的疾病表现的细胞机制，这在很大程度上阻碍了对受影响患者进行有效治疗的评估和实施。我们假设，线粒体RC功能障碍的细胞后果的药理学调制将提供有效的治疗RC功能障碍的常见亚组，无论个别致病原因。本提案的总体目标是阐明药理学药物调节RC功能障碍的代谢后果的机制，利用C。elegans动物模型提供。本提案的具体目的是：SA 1：确定是否在C. elegans将减弱或甚至逆转复合物I功能障碍的代谢后果;和SA 2：表征翻译C中“线粒体鸡尾酒”组分的体内后果。elegans复合物I功能障碍的动物模型。将分别在SA 1和SA 2中的RC复合物I突变体中研究直接调节PPAR/SIRT 1相关信号传导途径的5种药理学药物以及4个主要类别（维生素、抗氧化剂、复合物I翻译后激活剂和中间代谢调节剂）中的15种常见“线粒体鸡尾酒”组分。将进行生物信息学整合，以辨别每种药物对5种体内表型的总体疗效，以实现以下两个目标：（a）寿命;（B）通过表达微阵列分析进行代谢途径分析;（c）使用稳定同位素通过中间代谢途径的通量;（d）通过荧光显微镜检查进行线粒体膜电位;以及（e）通过荧光显微镜检查进行相对线粒体氧化剂负荷。潜在的可辨别的表型反应的机制，将探讨在体内功能分析复杂的I突变体窝藏RNA干扰诱导敲除个别的PPAR/SIRT 1途径基因。以这种方式，目前缺乏功效或危害的科学或临床证据的对人类RC疾病具有假定益处的药物可以在C中以低成本和高通量方式客观地和非侵入性地筛选。优雅这项转化研究可能提出特定的治疗靶点和潜在有效的药理学药物，以减轻人类线粒体疾病的全球后遗症。 公共卫生相关性：线粒体复合物I功能障碍发生在一组非常频繁，变化多样，并且在很大程度上无法治疗的遗传性疾病中，这些疾病困扰着所有年龄和种族。秀丽隐杆线虫提供了一个强大的遗传模型，以评估复杂的I功能障碍和治疗候选人的全球影响。这项转化研究可能证明有效的药理学疗法及其特定机制，以潜在地减轻人类线粒体疾病的继发性后果。

DESCRIPTION (provided by applicant): Dysfunction of complex I in the mitochondrial respiratory chain (RC) underlies an astonishingly frequent group of multi-systemic disorders that afflict all ages and ethnicities. A relative inability to objectively assess cellular mechanisms that mediate widely variable disease manifestations has largely prohibited evaluation and implementation of effective therapies in affected patients. We hypothesize that pharmacologic modulation of the cellular consequences of mitochondrial RC dysfunction will offer effective therapies for common subgroups of RC dysfunction, irrespective of individual pathogenic cause. The overall goal of this proposal is to elucidate mechanisms by which pharmacologic agents modulate the metabolic consequences of RC dysfunction, capitalizing on the inherent investigative advantages that the C. elegans animal model provides. The Specific Aims of this proposal are to: SA1: Determine whether pharmacologic modulation of the PPAR/SIRT1 pathway in C. elegans will attenuate, or even reverse, the metabolic consequences of complex I dysfunction; and SA2: Characterize in vivo consequences of "mitochondrial cocktail" components in a translational C. elegans animal model of complex I dysfunction. Five pharmacologic agents that directly modulate PPAR/SIRT1-related signaling pathways as well as fifteen common "mitochondrial cocktail" components in four major classes (vitamins, antioxidants, complex I post- translational activator, and intermediary metabolic modifiers) will be studied in RC complex I mutants in SA1 and SA2, respectively. Bioinformatics integration will be performed to discern the overall efficacy of each pharmacologic agent on 5 in vivo phenotypes for both aims: (a) lifespan; (b) metabolic pathway profiling by expression microarray analysis; (c) flux through intermediary metabolic pathways using stable isotopes; (d) mitochondrial membrane potential by fluorescence microscopy; and (e) relative mitochondrial oxidant burden by fluorescence microscopy. Mechanisms underlying discernible phenotypic responses will be explored by in vivo functional analyses in complex I mutants harboring RNA interference-induced knockdown for individual PPAR/SIRT1 pathway genes. In this manner, drugs having postulated benefit for human RC disease that currently lack scientific or clinical evidence of either efficacy or harm can be objectively and non- invasively screened in a low-cost and high-throughput fashion in C. elegans. This translational research may suggest specific therapeutic targets and potentially effective pharmacologic agents to mitigate the global sequelae of human mitochondrial disease. PUBLIC HEALTH RELEVANCE: Mitochondrial complex I dysfunction occurs in an astonishingly frequent, varied, and largely untreatable group of genetic disorders afflicting all ages and ethnicities. Caenorhabditis elegans offers a robust genetic model in which to assess the global impact of complex I dysfunction and therapeutic candidates. This translational research may demonstrate effective pharmacologic therapies, and their specific mechanisms, to potentially mitigate the secondary consequences of human mitochondrial disease.