Mouse Modeling of Leigh Disease and Complex I Assembly

Leigh 病和复合物 I 组装的小鼠模型

• 批准号: 7230077
• 负责人: Carl A. Pinkert
• 金额: $ 14.8万
• 依托单位: AUBURN UNIVERSITY AT AUBURN
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-15 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7230077
• 关键词: Mouse; Modeling; Leigh; Disease; Complex

DESCRIPTION (provided by applicant): Mitochondria provide energy for cells via oxidative phosphorylation (OXPHOS). The average cell contains hundreds to thousands of maternally-inherited mitochondria replete with their own mitochondrially-encoded genome. The interactions of mitochondrial and nuclear-encoded OXPHOS genes are critical in energy generation. Mutations in both nuclear- and mitochondrially-encoded OXPHOS genes have been identified but few targeted animal models exist. Defects within the OXPHOS genes are intimately associated with various metabolic diseases including severe childhood disorders including Leigh disease. Leigh disease is a neurodegenerative mitochondrial disorder characterized by a motor control loss and localized necrotic lesions. Leigh Disease can result from either mitochondrial or nuclear mutations - that cause devastating muscle and neurological degeneration between 3 months and 2 years of age along with a cascade of debilitating pathologies. This project focuses on developing genetically-engineered mouse models to characterize the role of specific OXPHOS genes and their involvement in OXPHOS pathways and disease progression. The majority of Leigh disease cases, whether acquired or sporadic, result from an insufficiency in production or function of particular electron transport proteins involved in oxidative phosphorylation. Various mutations in the NADH dehydrogenase-ubiquinone-FeS 4 (NDUFS4) gene have been associated with decreased Complex I activity in mitochondrial electron transport and Leigh disease. Specific phosphorylation of the NDUFS4 protein occurs in response to cAMP-dependent kinase activation and correlates with activation of Complex I. Conditional Cre-LoxP NDUFS4 heterozygous knockout mouse embryonic stem cells will be created to evaluate in vitro phenotype and to subsequently establish a homozygous knockout mouse model to examine in vivo phenotype and tissue specificity. A second embryonic stem cell line will be constructed to contain a heterozygous NDUFS4 point mutation. This will facilitate the creation of a homozygous point mutant mouse model that will address the specific mechanisms by which loss of a functional NDUFS4 protein can lead to Leigh disease. Phenotype will be identified by gross and histopathological analyses, in addition to specific measures of Complex I activity, cellular respiration and mitochondrial function. The two NDUFS4 mouse models will allow examination of mechanisms leading to Leigh disease. These models will further our understanding of mitochondrial function and Complex I assembly. As such, this project represents a first step toward developing therapeutic strategies and targeted interventions for debilitating mitochondrial disorders.

描述（由申请人提供）：线粒体通过氧化磷酸化（OXPHOS）为细胞提供能量。平均细胞包含数百至数千个母体亲属的线粒体充满自己的线粒体编码基因组。线粒体和核编码的OXPHOS基因的相互作用在能量产生中至关重要。已经鉴定出核和线粒体编码的OXPHOS基因的突变，但几乎没有靶向动物模型。 OXPHOS基因内的缺陷与包括严重儿童疾病在内的各种代谢疾病密切相关。 Leigh病是一种神经退行性线粒体疾病，其特征是运动控制损失和局部坏死病变。线粒体或核突变可能导致雷疾病 - 在3个月至2岁之间引起毁灭性的肌肉和神经系统变性以及一系列衰弱的病理。该项目着重于开发遗传工程的小鼠模型，以表征特定的Oxphos基因的作用及其参与OXPHOS途径和疾病进展。大多数Leigh疾病病例，无论是获得的还是零星的，是由于参与氧化磷酸化的特定电子转运蛋白的生产或功能不足而导致的。 NADH脱氢酶 - 泛素酮FES 4（NDUFS4）基因中的各种突变与线粒体电子转运和Leigh疾病中复合物I活性的降低有关。 NDUFS4蛋白的特异性磷酸化是响应cAMP依赖性激酶激活而发生的，并且与复杂的激活相关。条件CRE-LOXP NDUFS4杂合敲除小鼠胚胎干细胞将被创建以评估体外表型，并随后以结实的敲除小鼠模型，以建立VIVIV的探测和组织。将构建第二个胚胎干细胞系以包含杂合NDUFS4点突变。这将有助于创建纯合点突变小鼠模型，该模型将解决功能性NDUFS4蛋白损失会导致Leigh疾病的特定机制。除了复杂的I活性，细胞呼吸和线粒体功能的特定量度外，还将通过总体和组织病理学分析来鉴定表型。两种NDUFS4小鼠模型将允许检查导致Leigh疾病的机制。这些模型将进一步了解线粒体功能和复杂I组装。因此，该项目代表了制定治疗策略和针对性衰弱的线粒体疾病的第一步。