Rescue of a mitochondrial deficiency causing leber hereditary optic neuropathy

John Guy, Xiaoping Qi, Francesco Pallotti, Eric A. Schon, Giovanni Manfredi, Valerio Carelli, Andrea Martinuzzi, William W. Hauswirth, Alfred S. Lewin

Research output: Contribution to journalArticle

185 Citations (Scopus)

Abstract

A G to A transition at nucleotide 11778 in the ND4 subunit gene of complex I was the first point mutation in the mitochondrial genome linked to a human disease. It causes Leber Hereditary Optic Neuropathy, a disorder with oxidative phosphorylation deficiency. To overcome this defect, we made a synthetic ND4 subunit compatible with the "universal" genetic code and imported it into mitochondria by adding a mitochondrial targeting sequence. For detection we added a FLAG tag. This gene was inserted in an adeno-associated viral vector. The ND4FLAG protein was imported into the mitochondria of cybrids harboring the G11778A mutation, where it increased their survival rate threefold, under restrictive conditions that forced the cells to rely predominantly on oxidative phosphorylation to produce ATP. Since assays of complex I activity were normal in G11778A cybrids we focused on changes in ATP synthesis using complex I substrates. The G11778A cybrids showed a 60% reduction in the rate of ATP synthesis. Relative to mock-transfected G11778A cybrids, complemented G11778A cybrids showed a threefold increase in ATP synthesis, to a level indistinguishable from that in cybrids containing normal mitochondrial DNA. Restoration of respiration by allotopic expression opens the door for gene therapy of Leber Hereditary Optic Neuropathy.

Original languageEnglish
Pages (from-to)534-542
Number of pages9
JournalAnnals of Neurology
Volume52
Issue number5
DOIs
StatePublished - Nov 1 2002
Externally publishedYes

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Leber's Hereditary Optic Atrophy
Adenosine Triphosphate
Mitochondria
Genetic Code
Mitochondrial Diseases
Mitochondrial Genome
Oxidative Phosphorylation
Mitochondrial DNA
Point Mutation
Genetic Therapy
Genes
Respiration
Nucleotides
Mutation
Proteins

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Guy, J., Qi, X., Pallotti, F., Schon, E. A., Manfredi, G., Carelli, V., ... Lewin, A. S. (2002). Rescue of a mitochondrial deficiency causing leber hereditary optic neuropathy. Annals of Neurology, 52(5), 534-542. https://doi.org/10.1002/ana.10354

Rescue of a mitochondrial deficiency causing leber hereditary optic neuropathy. / Guy, John; Qi, Xiaoping; Pallotti, Francesco; Schon, Eric A.; Manfredi, Giovanni; Carelli, Valerio; Martinuzzi, Andrea; Hauswirth, William W.; Lewin, Alfred S.

In: Annals of Neurology, Vol. 52, No. 5, 01.11.2002, p. 534-542.

Research output: Contribution to journalArticle

Guy, J, Qi, X, Pallotti, F, Schon, EA, Manfredi, G, Carelli, V, Martinuzzi, A, Hauswirth, WW & Lewin, AS 2002, 'Rescue of a mitochondrial deficiency causing leber hereditary optic neuropathy', Annals of Neurology, vol. 52, no. 5, pp. 534-542. https://doi.org/10.1002/ana.10354
Guy, John ; Qi, Xiaoping ; Pallotti, Francesco ; Schon, Eric A. ; Manfredi, Giovanni ; Carelli, Valerio ; Martinuzzi, Andrea ; Hauswirth, William W. ; Lewin, Alfred S. / Rescue of a mitochondrial deficiency causing leber hereditary optic neuropathy. In: Annals of Neurology. 2002 ; Vol. 52, No. 5. pp. 534-542.
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