Selective elimination of mutant mitochondrial genomes as therapeutic strategy for the treatment of NARP and MILS syndromes

Gene Ther. 2008 Apr;15(7):516-23. doi: 10.1038/gt.2008.11. Epub 2008 Feb 7.


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 gammaH2ax 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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adenosine Triphosphate / analysis
  • Adenosine Triphosphate / biosynthesis
  • Adenoviridae / genetics
  • Blood Platelets
  • Cell Line, Tumor
  • Cell Proliferation
  • Cell Respiration
  • Coculture Techniques
  • DNA, Mitochondrial / genetics*
  • Deoxyribonucleases, Type II Site-Specific / genetics*
  • Deoxyribonucleases, Type II Site-Specific / metabolism
  • Galactose / metabolism
  • Gene Deletion
  • Genetic Engineering
  • Genetic Markers
  • Genetic Therapy / methods*
  • Genetic Vectors / administration & dosage
  • Genome, Mitochondrial
  • Humans
  • Lactic Acid / metabolism
  • Mitochondrial Diseases / genetics
  • Mitochondrial Diseases / metabolism
  • Mitochondrial Diseases / therapy*
  • Mutation*
  • Oxidative Phosphorylation
  • Transduction, Genetic / methods*


  • DNA, Mitochondrial
  • Genetic Markers
  • Lactic Acid
  • Adenosine Triphosphate
  • CCCGGG-specific type II deoxyribonucleases
  • Deoxyribonucleases, Type II Site-Specific
  • Galactose