The optimized allotopic expression of ND1 or ND4 genes restores respiratory chain complex I activity in fibroblasts harboring mutations in these genes
The optimized allotopic expression of ND1 or ND4 genes restores respiratory chain complex I activity in fibroblasts harboring mutations in these genes
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DOI：
10.1016/j.bbamcr.2008.04.018
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发表时间：
2008-10-01
影响因子：
5.1
通讯作者：
Corral-Debrinski, Marisol
中科院分区：
文献类型：
--
作者：
Bonnet, Crystel;Augustin, Sebastien;Corral-Debrinski, Marisol
Leber遗传性视神经病变（LHON）是第一个被鉴定的母系遗传性线粒体疾病，现在被认为是最普遍的线粒体疾病。LHON患者携带线粒体DNA（mtDNA）突变。在大约90%的病例中，所涉及的基因编码呼吸链复合物I的蛋白质。尽管分子基础是已知的，因为20年来，几乎所有仍然是关于病理生理学和治疗做。在这项研究中，我们报告了一个严重的减少复杂的I活性培养的皮肤成纤维细胞分离两个LHON患者窝藏ND4或ND1基因突变。最重要的是，我们能够可持续地恢复（a）在半乳糖上生长的能力，（B）ATP合成速率和（c）最初在这些细胞中受损的复合物I活性。我们的策略包括迫使核编码的ND1和ND4基因的mRNA定位到线粒体表面。观察到的呼吸链缺陷的拯救是可能的，通过离散量的杂交mRNA和融合蛋白，证明其线粒体输入的效率。因此，我们在此证实了位于细胞器中的两个线粒体基因，优化的异位表达方法代表了一种强大的工具，最终可以应用于LHON的人类治疗。(c)2008 Elsevier BY，保留所有权利。
Leber's Hereditary Optic Neuropathy (LHON) was the first maternally inherited mitochondrial disease identified and is now considered the most prevalent mitochondrial disorder. LHON patients harbor mutations in mitochondrial DNA (mtDNA). In about 90% of cases, the genes involved encode proteins of the respiratory chain complex I. Even though the molecular bases are known since 20 years almost all remains to be done regarding physiopathology and therapy. In this study, we report a severe decrease of complex I activity in cultured skin fibroblasts isolated from two LHON patients harboring mutations in ND4 or ND1 genes. Most importantly, we were able to restore sustainably (a) the ability to grow on galactose, (b) the ATP synthesis rate and (c) the complex I activity, initially impaired in these cells. Our strategy consisted of forcing mRNAs from nuclearly-encoded ND1 and ND4 genes to localize to the mitochondrial surface. The rescue of the respiratory chain defect observed was possible by discreet amounts of hybrid mRNAs and fusion proteins demonstrating the efficiency of their mitochondrial import. Hence, we confirmed here for two mitochondrial genes located in the organelle that the optimized allotopic expression approach represents a powerful tool that could ultimately be applied in human therapy for LHON. (c) 2008 Elsevier BY, All rights reserved.