Translational Genomic Study of Mitochondrial Complex I Dysfunction in C. elegans

线虫线粒体复合物 I 功能障碍的转化基因组研究

• 批准号: 7126382
• 负责人: MARNI J FALK
• 金额: $ 13.39万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-30 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7126382
• 关键词: Caenorhabditis elegans; RNA interference; alternatives to animals in research; anesthesia; biochemical evolution; cellular respiration; developmental genetics; disease /disorder etiology; functional /structural genomics; gene expression; genetic models; genome; helminth genetics; inborn metabolism disorder; longevity; microarray technology; mitochondrial disease /disorder; oxidative phosphorylation; pharmacogenetics

DESCRIPTION (provided by applicant): Mitochondrial oxidative phosphorylation dysfunction is one of the most common inborn errors of metabolism. However, it is difficult to diagnose and its genetic etiology is often unclear. This proposal's immediate goal is to exploit the nematode C. elegans as a translational model system to enhance the genetic study of mitochondrial disease. The long-term objective is to improve diagnostic capabilities for the heterogeneous subset of human patients with genetic-based mitochondrial disease. Complex I is the largest and most commonly implicated mitochondrial respiratory chain (MRC) complex in human mitochondrial disease. This project will clarify contributions of structural subunits of complex I to integrated mitochondrial function in C. elegans. The proposed approach is based on two hypotheses: 1)Protein components of MRC complexes conserved between C. elegans and humans are critical to proper MRC function; and 2)Consistent patterns of compensatory responses in gene expression occur in the nuclear genome when MRC function is impaired. Specific aims for this proposal are to: 1) Determine which of the evolutionary conserved, nuclear-encoded subunits of complex I are integral to mitochondrial function in C. elegans, and 2) Identify a representative gene expression-based pattern within biologically relevant pathways that is indicative of MRC dysfunction. An RNA interference feeding approach will be used to inhibit expression of conserved complex I subunit nuclear genes. The effect of this disruption will be characterized on mitochondrial function, as assessed by oxidative phosphorylation capacity; on whole organism function as assessed by anesthetic sensitivity, lifespan, and growth rate; and on cellular compensatory mechanisms as assessed by microarray analysis of the entire C. elegans nuclear genome using gene set enrichment analysis of selected biologically-relevant gene clusters. RELEVANCE: Despite increased awareness of the important role mitochondria play in common diseases, such as type II diabetes, to less common inborn errors of metabolism, there is limited ability to pursue definitive genetic diagnoses. C. elegans presents an unparalleled ability to correlate findings of mitochondrial dysfunction to mutations in specific genes, and to suggest gene patterns that may be used to screen or diagnose human mitochondrial disease. Understanding specific genetic causes of mitochondrial disease will provide the basis for rational clinical diagnosis, treatment, and perhaps, cure.

描述（由申请人提供）： 线粒体氧化磷酸化功能障碍是最常见的代谢误差之一。但是，很难诊断，其遗传病因通常不清楚。该提议的近期目标是利用线虫秀丽隐杆线虫作为转化模型系统，以增强线粒体疾病的遗传研究。长期目标是改善基于遗传的线粒体疾病的人类患者的异质子集的诊断能力。复合体I是人线粒体疾病中最大，最常见的线粒体呼吸链（MRC）复合物。该项目将阐明复合物I的结构亚基对秀丽隐杆线虫中的线粒体功能的贡献。提出的方法基于两个假设：1）秀丽隐杆线虫和人之间保守的MRC复合物的蛋白质成分对于适当的MRC功能至关重要； 2）当MRC功能受损时，基因表达中补偿性反应的一致模式发生。该提案的具体目的是：1）确定复合物I的进化保守的，核编码的亚基是秀丽隐杆线虫中线粒体功能不可或缺的一部分，以及2）确定一种在生物学上相关的途径中，这表明MRC功能障碍。 RNA干扰进食方法将用于抑制保守的复合物I亚基核基因的表达。通过氧化磷酸化能力评估，这种破坏的影响将在线粒体功能上进行表征。通过麻醉敏感性，寿命和生长速率评估整个生物体的功能；以及通过对整个秀丽隐杆线虫核基因组进行微阵列分析的细胞补偿机制，使用选定的与生物学相关的基因簇的基因集富集分析。相关性：尽管对线粒体在常见疾病（例如II型糖尿病）中的重要作用提高了对代谢的先天错误，但追求确定的遗传诊断的能力有限。秀丽隐杆线虫具有将线粒体功能障碍与特定基因突变相关联的无与伦比的能力，并提出了可用于筛查或诊断人类线粒体疾病的基因模式。了解线粒体疾病的特定遗传原因将为合理的临床诊断，治疗以及治愈提供基础。