mitoTALEN reduces the mutant mtDNA load in neurons
mitoTALEN reduces the mutant mtDNA load in neurons
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DOI：
10.1016/j.omtn.2024.102132
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发表时间：
2024-02-16
期刊：
影响因子：
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通讯作者：
Moraes，Carlos T.
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文献类型：
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作者：
Bacman，Sandra R.;Barrera-Paez，Jose Domingo;Moraes，Carlos T.
线粒体DNA内的突变经常引起严重的脑病。鉴于这些mtDNA缺陷中的大多数以异质状态存在，我们利用靶向线粒体的TALEN（mitoTALEN）的精确性来选择性地消除在线粒体tRNA丙氨酸基因（m.5024C>T）中具有异质突变的小鼠模型的CNS内的突变mtDNA。这种靶向方法是通过使用AAV-PHP.eB和神经元特异性突触蛋白启动子来实现的，用于mitoTALEN的有效神经元递送和表达。我们发现，大多数中枢神经系统区域被有效地转导，并表现出显着减少突变mtDNA。这种减少伴随着线粒体tRNA丙氨酸水平的增加，这是由m.5024C>T突变急剧减少。这些结果表明，脑靶向基因编辑可以有效地减少CNS突变体的体内mtDNA，为线粒体脑病患者的临床试验铺平了道路。
Mutations within mtDNA frequently give rise to severe encephalopathies. Given that a majority of these mtDNA defects exist in a heteroplasmic state, we harnessed the precision of mitochondrial-targeted TALEN (mitoTALEN) to selectively eliminate mutant mtDNA within the CNS of a murine model harboring a heteroplasmic mutation in the mitochondrial tRNA alanine gene (m.5024C>T). This targeted approach was accomplished by the use of AAV-PHP.eB and a neuron-specific synapsin promoter for effective neuronal delivery and expression of mitoTALEN. We found that most CNS regions were effectively transduced and showed a significant reduction in mutant mtDNA. This reduction was accompanied by an increase in mitochondrial tRNA alanine levels, which are drastically reduced by the m.5024C>T mutation. These results showed that mitochondrial-targeted gene editing can be effective in reducing CNS-mutant mtDNAin vivo, paving the way for clinical trials in patients with mitochondrial encephalopathies.