Department of Neurology, University of Miami School of Medicine, Miami, FL, USA.
Gene Ther. 2012 Nov;19(11):1101-6. doi: 10.1038/gt.2011.196. Epub 2011 Dec 1.
Mitochondrial diseases are frequently caused by heteroplasmic mitochondrial DNA (mtDNA) mutations. As these mutations express themselves only at high relative ratios, any approach able to manipulate mtDNA heteroplasmy can potentially be curative. In this study, we developed a system to manipulate mtDNA heteroplasmy in all skeletal muscles from neonate mice. We selected muscle because it is one of the most clinically affected tissues in mitochondrial disorders. A mitochondria-targeted restriction endonuclease (mito-ApaLI) expressed from AAV9 particles was delivered either by intraperitoneal or intravenous injection in neonate mice harboring two mtDNA haplotypes, only one of which was susceptible to ApaLI digestion. A single injection was able to elicit a predictable and marked change in mtDNA heteroplasmy in all striated muscles analyzed, including heart. No health problems or reduction in mtDNA levels were observed in treated mice, suggesting that this approach could have clinical applications for mitochondrial myopathies.
线粒体疾病通常由异质体线粒体 DNA(mtDNA)突变引起。由于这些突变仅在相对高的比例下表达,因此任何能够操纵 mtDNA 异质性的方法都可能具有治疗效果。在这项研究中,我们开发了一种在新生小鼠所有骨骼肌中操纵 mtDNA 异质性的系统。我们选择肌肉是因为它是线粒体疾病中最受临床影响的组织之一。一种靶向线粒体的限制性内切酶(mito-ApaLI)由 AAV9 颗粒表达,通过腹腔内或静脉内注射递送至携带两种 mtDNA 单倍型的新生小鼠中,其中只有一种易受 ApaLI 消化。单次注射能够引起所有横纹肌(包括心脏)中 mtDNA 异质性的可预测且显著变化。在治疗的小鼠中未观察到健康问题或 mtDNA 水平降低,表明该方法可能对线粒体肌病具有临床应用价值。