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Organ-specific shifts in mtDNA heteroplasmy following systemic delivery of a mitochondria-targeted restriction endonuclease

Gene Therapy volume 17, pages 713–720 (2010)Cite this article

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Abstract

Most pathogenic mtDNA mutations are heteroplasmic and there is a clear correlation between high levels of mutated mtDNA in a tissue and pathology. We have found that in vivo double-strand breaks (DSBs) in mtDNA lead to digestion of cleaved mtDNA and replication of residual mtDNA. Therefore, if DSB could be targeted to mutations in mtDNA, mutant genomes could be eliminated and the wild-type mtDNA would repopulate the cells. This can be achieved by using mitochondria-targeted restriction endonucleases as a means to degrade specific mtDNA haplotypes in heteroplasmic cells or tissues. In this work, we investigated the potential of systemic delivery of mitochondria-targeted restriction endonucleases to reduce the proportion of mutant mtDNA in specific tissues. Using the asymptomatic NZB/BALB mtDNA heteroplasmic mouse as a model, we found that a mitochondria-targeted ApaLI (that cleaves BALB mtDNA at a single site and does not cleave NZB mtDNA) increased the proportion of NZB mtDNA in target tissues. This was observed in heart, using a cardiotropic adeno-associated virus type-6 (AAV6) and in liver, using the hepatotropic adenovirus type-5 (Ad5). No mtDNA depletion or loss of cytochrome c oxidase activity was observed in any of these tissues. These results show the potential of systemic delivery of viral vectors to specific organs for the therapeutic application of mitochondria-targeted restriction enzymes in mtDNA disorders.

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Acknowledgements

We are grateful to James M Allen and Jeffrey S Chamberlain for help preparing high titers of AAV6 and advice. This work was supported by PHS grant EY10804, NS041777, CA85700 and the Muscular Dystrophy Association to CTM. This project had the support of the core grant 5P30EY014801-07.

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  1. S R Bacman and S L Williams: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Neurology, University of Miami School of Medicine, Miami, FL, USA

    S R Bacman, S L Williams, S Garcia & C T Moraes

  2. Department of Cell Biology and Anatomy, University of Miami School of Medicine, Miami, FL, USA

    C T Moraes

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  1. S R Bacman
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Correspondence to C T Moraes.

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Bacman, S., Williams, S., Garcia, S. et al. Organ-specific shifts in mtDNA heteroplasmy following systemic delivery of a mitochondria-targeted restriction endonuclease. Gene Ther 17, 713–720 (2010). https://doi.org/10.1038/gt.2010.25

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