Selective elimination of mutant mitochondrial genomes as therapeutic strategy for the treatment of NARP and MILS syndromes
Selective elimination of mutant mitochondrial genomes as therapeutic strategy for the treatment of NARP and MILS syndromes
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DOI：
10.1038/gt.2008.11
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发表时间：
2008-04-01
期刊：
影响因子：
5.1
通讯作者：
Wilson, G. L.
中科院分区：
文献类型：
--
作者：
Alexeyev, M. F.;Venediktova, N.;Wilson, G. L.
线粒体疾病并不罕见，可能是由核和线粒体DNA（mtDNA）突变引起的。目前，只有姑息疗法可用于这些疾病，并在开发有效的治疗方案的兴趣很高。在这里，我们证明了在细胞异质性的T8993G突变，这是一个原因，NARP和MILS综合征，感染腺病毒，它编码的尿道靶向R。XmaI限制性内切酶，导致突变mtDNA的选择性破坏。这种破坏以时间和剂量依赖性的方式进行，并导致细胞的耗氧率和ATP产生率显着增加。R.线粒体的XmaI伴随着利用半乳糖作为唯一碳源的能力的提高，这是氧化磷酸化能力的替代指标。同时，这些细胞产生乳酸的速率降低，这是线粒体功能障碍的标志。我们进一步证明，磷酸化的P53和γ H2ax蛋白，核DNA损伤的标志物的水平，不改变重组腺病毒感染，表明核DNA损伤的情况下，该技术的相对安全。最后，一些所提出的方法的优点和局限性进行了讨论。
Mitochondrial diseases are not uncommon, and may result from mutations in both nuclear and mitochondrial DNA (mtDNA). At present, only palliative therapies are available for these disorders, and interest in the development of efficient treatment protocols is high. Here, we demonstrate that in cells heteroplasmic for the T8993G mutation, which is a cause for the NARP and MILS syndromes, infection with an adenovirus, which encodes the mitochondrially targeted R. XmaI restriction endonuclease, leads to selective destruction of mutant mtDNA. This destruction proceeds in a time-and dose-dependent manner and results in cells with significantly increased rates of oxygen consumption and ATP production. The delivery of R. XmaI to mitochondria is accompanied by improvement in the ability to utilize galactose as the sole carbon source, which is a surrogate indicator of the proficiency of oxidative phosphorylation. Concurrently, the rate of lactic acid production by these cells, which is a marker of mitochondrial dysfunction, decreases. We further demonstrate that levels of phosphorylated P53 and gamma H2ax proteins, markers of nuclear DNA damage, do not change in response to infection with recombinant adenovirus indicating the absence of nuclear DNA damage and the relative safety of the technique. Finally, some advantages and limitations of the proposed approach are discussed.