Pre-clinical studies of novel mitochondrial gene therapies

 描述（由申请人提供）：线粒体疾病是一种常见的、具有破坏性的疾病，预后极差。已经提出了涉及从细胞核中异位表达重新编码的线粒体基因的基因疗法，然而，这种方法的可行性仍然存在争议。我们已经发现，这种方法的主要技术障碍，限制了一种新的基因治疗的发展是竞争的内源性突变蛋白质的复杂性和这些蛋白质的疏水性。我们已经发现了一种新的线粒体翻译抑制（TLI）方法，以防止线粒体内的突变蛋白的表达。我们建议测试线粒体TLI与靶向编码RNA至线粒体的新治疗方法的组合。这种方法直接解决了这两个技术障碍，使用新的和以前未经测试的方法。任何旨在开发新的线粒体基因疗法的认真研究都需要在可行的遗传系统中充分表征致病性内源性线粒体突变，其中可以在体内证明和优化可行性。我们提出了一个严格的测试线粒体TLI和神经靶向RNA的表达，使用几个生化和表型功能测定在一个良好的特征动物模型系统。

 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.