INSERM U613; Université de Brest, Faculté de Médecine et des Sciences de la Santé; Etablissement Français du Sang (EFS) Bretagne; Centre Hospitalier Régional Universitaire Brest, Hop Morvan, Laboratoire de Génétique Moléculaire, F-29200 Brest, France.
Proc Natl Acad Sci U S A. 2011 Jul 19;108(29):11989-94. doi: 10.1073/pnas.1101478108. Epub 2011 Jun 29.
Due to the lack of relevant animal models, development of effective treatments for human mitochondrial diseases has been limited. Here we establish a rapid, yeast-based assay to screen for drugs active against human inherited mitochondrial diseases affecting ATP synthase, in particular NARP (neuropathy, ataxia, and retinitis pigmentosa) syndrome. This method is based on the conservation of mitochondrial function from yeast to human, on the unique ability of yeast to survive without production of ATP by oxidative phosphorylation, and on the amenability of the yeast mitochondrial genome to site-directed mutagenesis. Our method identifies chlorhexidine by screening a chemical library and oleate through a candidate approach. We show that these molecules rescue a number of phenotypes resulting from mutations affecting ATP synthase in yeast. These compounds are also active on human cybrid cells derived from NARP patients. These results validate our method as an effective high-throughput screening approach to identify drugs active in the treatment of human ATP synthase disorders and suggest that this type of method could be applied to other mitochondrial diseases.
由于缺乏相关的动物模型,人类线粒体疾病的有效治疗方法的发展受到了限制。在这里,我们建立了一种快速的酵母基础检测方法,用于筛选针对影响 ATP 合酶的人类遗传性线粒体疾病的药物,特别是 NARP(神经病、共济失调和视网膜色素变性)综合征。该方法基于从酵母到人线粒体功能的保守性、酵母在没有氧化磷酸化产生 ATP 的情况下生存的独特能力,以及酵母线粒体基因组对定点诱变的适应性。我们的方法通过筛选化学文库鉴定了洗必泰,并通过候选方法鉴定了油酸。我们表明,这些分子可挽救影响酵母中 ATP 合酶的突变引起的多种表型。这些化合物对源自 NARP 患者的人杂种细胞也具有活性。这些结果验证了我们的方法作为一种有效的高通量筛选方法,用于鉴定可有效治疗人类 ATP 合酶疾病的药物,并表明这种类型的方法可应用于其他线粒体疾病。