Pharmacologic treatment of mitochondrial complex I dysfunction in C. elegans

线虫线粒体复合物 I 功能障碍的药物治疗

基本信息

批准号：
8484859
负责人：
MARNI J FALK
金额：
$ 37.83万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-27 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8484859
关键词：
Acetylcysteine Adult Affect Age Agmatine Amino Acids Animal Model Animals Antioxidants Arginine Attenuated Biochemical Bioinformatics Biological Caenorhabditis elegans Carnitine Citric Acid Cycle Clinical Coenzyme Q10 Complex Creatine Development Diagnosis Disease Dose Drug Prescriptions Ethnic Origin Evaluation Fenofibrate Fluorescence Microscopy Functional disorder Gases Genes Genetic Genetic Models Goals Hereditary Disease Human Hydroxybutyrates Impairment Individual Leucovorin Life Longevity Mammals Mediating Membrane Potentials Metabolic Metabolic Pathway Microarray Analysis Mitochondria Mitochondrial Diseases Modeling Molecular Profiling Nematoda Nicotinic Acids Organ failure Organism Oxidants Pathway interactions Patients Peroxisome Proliferator-Activated Receptors Pharmaceutical Preparations Phenotype Physiological Prevalence Probucol RNA Interference Relative (related person)Respiratory Chain Resveratrol Signal Pathway Subgroup Symptoms System Therapeutic Thiamine Thioctic Acid Translational Research Vitamin E Vitamins Work base cost effective therapy feeding in vivo mitochondrial membrane mutant new therapeutic target orotate oxidant stress public health relevance response rosiglitazone sirtinol stable isotope therapeutic target

项目摘要

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.

描述（由申请人提供）：线粒体呼吸链（RC）中复合物 I 的功能障碍是一组令人惊讶的常见多系统疾病的基础，这些疾病困扰着所有年龄和种族的人。相对无法客观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的有效治疗的评估和实施。我们假设，无论个体致病原因如何，对线粒体 RC 功能障碍的细胞后果进行药理学调节将为常见的 RC 功能障碍亚组提供有效的治疗方法。该提案的总体目标是利用秀丽隐杆线虫动物模型提供的固有研究优势，阐明药物调节 RC 功能障碍代谢后果的机制。该提案的具体目标是： SA1：确定秀丽隐杆线虫中 PPAR/SIRT1 通路的药理调节是否会减弱甚至逆转复合物 I 功能障碍的代谢后果； SA2：表征复杂 I 功能障碍的转化秀丽隐杆线虫动物模型中“线粒体鸡尾酒”成分的体内后果。 RC 复合物将研究直接调节 PPAR/SIRT1 相关信号通路的五种药物制剂以及四大类（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 15 种常见的“线粒体鸡尾酒”成分I 分别为 SA1 和 SA2 突变体。将进行生物信息学整合，以辨别每种药物对 5 种体内表型的总体功效，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) 通过荧光显微镜观察线粒体膜电位； (e) 通过荧光显微镜观察线粒体的相对氧化剂负荷。将通过对复杂 I 突变体进行体内功能分析来探索可辨别表型反应的机制，该突变体含有 RNA 干扰诱导的单个 PPAR/SIRT1 途径基因的敲低。以这种方式，可以在秀丽隐杆线虫中以低成本和高通量的方式客观且非侵入性地筛选目前缺乏功效或危害的科学或临床证据的假定对人类RC疾病有益的药物。这项转化研究可能会提出特定的治疗靶点和潜在有效的药物制剂，以减轻人类线粒体疾病的全球后遗症。