PMID- 25342621 OWN - NLM STAT- MEDLINE DCOM- 20150126 LR - 20190327 IS - 1552-5783 (Electronic) IS - 0146-0404 (Print) IS - 0146-0404 (Linking) VI - 55 IP - 12 DP - 2014 Oct 23 TI - LHON gene therapy vector prevents visual loss and optic neuropathy induced by G11778A mutant mitochondrial DNA: biodistribution and toxicology profile. PG - 7739-53 LID - 10.1167/iovs.14-15388 [doi] AB - PURPOSE: To demonstrate safety and efficacy of allotopic human ND4 for treatment of a Leber's hereditary optic neuropathy (LHON) mouse model harboring the G11778A mitochondrial mutation. METHODS: We induced LHON in mice by intravitreal injection of mutant (G11778A) human ND4 DNA, responsible for most cases of LHON, that was directed to mitochondria using an AAV2 vector to which we appended a mitochondrial targeting sequence to the VP2 capsid. We then attempted rescue of visual loss using our test article (ScAAV2-P1ND4v2) containing a synthetic nuclear encoded G11778G ND4 gene that was allotopically expressed. Control mice either were uninjected or received AAV2-GFP or AAV2-mCherry. We performed RT-PCR and confocal microscopy at 2 weeks post injection. Pattern electroretinograms (PERGs), spectral-domain optical coherence tomography (SD-OCT), histology, and transmission electron microscopy (TEM) were performed. For toxicology and biodistribution studies, the test article was administered intravitreally to rats and rhesus macaques at different doses. RESULTS: Mutant and wild-type ND4 were efficiently expressed in the mitochondria of retinal ganglion cells (RGCs). Visual function assessed by serial PERGs and retinal structure by serial SD-OCT showed a significant rescue by the test article. Histology and ultrastructural analysis confirmed that loss of RGCs and demise of axons was prevented by ScAAV2-P1ND4v2. Rat and nonhuman primate biodistribution studies showed that vector spread outside the injected eye into spleen and lymph nodes was minimal. Histopathology of tissues and organs including the eyes was comparable to that of uninfected and saline-injected eyes. CONCLUSIONS: Allotopically expressed wild-type ND4 prevents the phenotype induced by G11778A mitochondrial DNA with a toxicology profile acceptable for testing in a phase I clinical trial. CI - Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc. FAU - Koilkonda, Rajeshwari AU - Koilkonda R AD - Bascom Palmer Eye Institute, University of Miami, Miller School of Medicine, Miami, Florida, United States. FAU - Yu, Hong AU - Yu H AD - Bascom Palmer Eye Institute, University of Miami, Miller School of Medicine, Miami, Florida, United States. FAU - Talla, Venu AU - Talla V AD - Bascom Palmer Eye Institute, University of Miami, Miller School of Medicine, Miami, Florida, United States. FAU - Porciatti, Vittorio AU - Porciatti V AD - Bascom Palmer Eye Institute, University of Miami, Miller School of Medicine, Miami, Florida, United States. FAU - Feuer, William J AU - Feuer WJ AD - Bascom Palmer Eye Institute, University of Miami, Miller School of Medicine, Miami, Florida, United States. FAU - Hauswirth, William W AU - Hauswirth WW AD - Department of Ophthalmology, University of Florida, College of Medicine, Gainesville, Florida, United States. FAU - Chiodo, Vince AU - Chiodo V AD - Department of Ophthalmology, University of Florida, College of Medicine, Gainesville, Florida, United States. FAU - Erger, Kirsten E AU - Erger KE AD - Department of Pediatrics, University of Florida, College of Medicine, Gainesville, Florida, United States. FAU - Boye, Sanford L AU - Boye SL AD - Department of Ophthalmology, University of Florida, College of Medicine, Gainesville, Florida, United States. FAU - Lewin, Alfred S AU - Lewin AS AD - Department of Molecular Genetics and Microbiology, University of Florida, College of Medicine, Gainesville, Florida, United States. FAU - Conlon, Thomas J AU - Conlon TJ AD - Department of Pediatrics, University of Florida, College of Medicine, Gainesville, Florida, United States. FAU - Renner, Lauren AU - Renner L AD - Oregon National Primate Research Center, Oregon Health and Science University, Beaverton, Oregon, United States. FAU - Neuringer, Martha AU - Neuringer M AD - Oregon National Primate Research Center, Oregon Health and Science University, Beaverton, Oregon, United States. FAU - Detrisac, Carol AU - Detrisac C AD - Charles River Pathology Associates-Illinois, Chicago, Illinois, United States. FAU - Guy, John AU - Guy J AD - Bascom Palmer Eye Institute, University of Miami, Miller School of Medicine, Miami, Florida, United States. LA - eng GR - P51 OD011092/OD/NIH HHS/United States GR - P30 EY014801/EY/NEI NIH HHS/United States GR - R01EY01714/EY/NEI NIH HHS/United States GR - R01EY123555/EY/NEI NIH HHS/United States GR - R01 EY019077/EY/NEI NIH HHS/United States GR - R24 EY018600/EY/NEI NIH HHS/United States GR - R24EY018600/EY/NEI NIH HHS/United States GR - P30 EY021721/EY/NEI NIH HHS/United States GR - P51OD011092/OD/NIH HHS/United States GR - P30EY014801/EY/NEI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20141023 PL - United States TA - Invest Ophthalmol Vis Sci JT - Investigative ophthalmology & visual science JID - 7703701 RN - 0 (DNA, Mitochondrial) RN - 0 (NADH dehydrogenase subunit 4) RN - EC 1.6.99.3 (NADH Dehydrogenase) SB - IM MH - Animals MH - Axons/ultrastructure MH - Blindness/genetics/*therapy MH - DNA, Mitochondrial/*genetics MH - Disease Models, Animal MH - Electroretinography MH - Genetic Therapy/adverse effects/*methods MH - Genetic Vectors/genetics MH - Macaca mulatta MH - Mice MH - Microscopy, Electron, Transmission MH - Mitochondria/*genetics/metabolism MH - NADH Dehydrogenase/genetics/*metabolism MH - Optic Atrophy, Hereditary, Leber/genetics/physiopathology/*therapy MH - Rats MH - Retinal Ganglion Cells/metabolism/ultrastructure MH - Tomography, Optical Coherence PMC - PMC4249950 OTO - NOTNLM OT - LHON OT - gene therapy OT - mitochondria EDAT- 2014/10/25 06:00 MHDA- 2015/01/27 06:00 PMCR- 2015/06/01 CRDT- 2014/10/25 06:00 PHST- 2014/10/25 06:00 [entrez] PHST- 2014/10/25 06:00 [pubmed] PHST- 2015/01/27 06:00 [medline] PHST- 2015/06/01 00:00 [pmc-release] AID - iovs.14-15388 [pii] AID - 10.1167/iovs.14-15388 [doi] PST - epublish SO - Invest Ophthalmol Vis Sci. 2014 Oct 23;55(12):7739-53. doi: 10.1167/iovs.14-15388.