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Laboratory Sciences
Feburary 2007

Use of Mitochondrial Antioxidant Defenses for Rescue of Cells With a Leber Hereditary Optic Neuropathy–Causing Mutation

Author Affiliations

Author Affiliations: Departments of Ophthalmology (Drs Qi, Sun, Hauswirth, and Guy), Molecular Genetics and Microbiology (Drs Hauswirth and Lewin), and Neurology (Dr Guy), College of Medicine, University of Florida, Gainesville.

Arch Ophthalmol. 2007;125(2):268-272. doi:10.1001/archopht.125.2.268

Objective  To explore a treatment paradigm for Leber hereditary optic neuropathy (LHON), we augmented mitochondrial antioxidant defenses to rescue cells with the G11778A mutation in mitochondrial DNA.

Methods  Cells homoplasmic for the G11778A mutation in mitochondrial DNA were infected with an adeno-associated viral vector containing the human mitochondrial superoxide dismutase (SOD2) gene. Control cells were infected with an adeno-associated viral (AAV) vector expressing the green fluorescent protein (GFP). Two days later, the high-glucose culture medium was exchanged for a glucose-free medium containing galactose. After 1 or 2days, cellular production of superoxide was assessed using the fluorescent probe dihydroethidium, and we used TUNEL (terminal deoxynucleotidyl transferase–mediated biotin–deoxyuridine triphosphate nick-end labeling) staining to detect apoptotic nuclei. The effect of SOD2 on LHON cell survival was quantitated after 2 or 3 days.

Results  Comparisons of AAV-SOD2–infected LHON cells relative to control cells infected with AAV–green fluorescent protein showed increased expression of mitochondrial SOD that attenuated superoxide-induced fluorescence by 26% (P = .003) and suppressed TUNEL-induced fluorescence by 21% (P = .048)after 2 days of growth in galactose medium, when cell survival increased by 25% (P=.05). After 3 days in galactose medium, SOD2 increased LHON survival by 89% (P = .006) relative to controls.

Conclusion  Protection against mitochondrial oxidative stress may be useful for treatment of LHON.

Clinical Relevance  Gene therapy with antioxidant genes may protect patients with LHON against visual loss.