Pre-clinical studies of novel mitochondrial gene therapies

 描述(申请人提供)：线粒体疾病是一种常见的破坏性疾病，预后极差。已经提出了涉及从细胞核异位表达重新编码的线粒体基因的基因治疗方法，然而，这种方法的可行性仍然存在争议。我们发现，限制新型基因治疗发展的主要技术障碍是来自复合体中内源突变蛋白的竞争以及这些蛋白的疏水性。我们发现了一种新的线粒体翻译抑制(TLI)方法来防止突变蛋白在线粒体内的表达。我们建议测试线粒体TLI与一种针对线粒体编码RNA的新治疗方法的组合。这种方法使用新的和以前未经测试的方法直接解决了两个技术障碍。任何旨在开发新的线粒体基因疗法的严肃研究都需要一个具有良好特征的、致病的、内源性的线粒体突变，在一个可在体内证明和优化可行性的遗传系统中。我们提出了一项严格的线粒体TLI和线粒体靶向RNA表达的测试，使用了几种生化和表型功能分析，在一个特征良好的动物模型系统中。

 DESCRIPTION (provided by applicant): Mitochondrial diseases are common and devastating conditions with an extremely poor prognosis. Gene therapies have been proposed involving allotopic expression of recoded mitochondrial genes from the nucleus, however, the viability of such an approach remains controversial. We have discovered that the major technical hurdles to such an approach that limit the development of a novel gene therapy are competition from the endogenous mutant protein in the complex and the hydrophobicity of these proteins. We have discovered a novel mitochondrial translation inhibition (TLI) approach to prevent expression of the mutant protein within mitochondria. We propose to test the combination of mitochondrial TLI with a novel therapy approach that targets coding RNAs to mitochondria. Such an approach directly addresses both technical hurdles using novel and previously untested methods. Any serious investigation aimed at developing a novel mitochondrial gene therapy would require a well-characterized, pathogenic, endogenous mitochondrial mutation in an amenable genetic system where feasibility can be demonstrated and optimized in vivo. We propose a rigorous test of mitochondrial TLI and mitochondrial-targeted RNA expression using several biochemical and phenotypic assays of function in a well-characterized animal model system.