title: Gene therapy with mitochondrial heat shock protein 70 suppresses visual loss and optic atrophy in experimental autoimmune encephalomyelitis.
abstract: OBJECTIVE
  To rescue visual loss and optic neuropathy in experimental autoimmune encephalomyelitis (EAE).
  
  
  METHODS
  Encephalomyelitis was induced in mice that received intravitreal injections of AAV2-mtHSP70Flag or AAV2-Cox8-mCherry. Additional mice were injected with AAV2-Cox8-mCherry, but not sensitized for EAE. Visual function was assessed by pattern electroretinograms (PERG) at 1, 3, and 6 months post injection (MPI). Optical coherence tomography (OCT) evaluated the thickness of the inner plexiform layer + nerve fiber layers at 1, 3, and 6 MPI. Retinas and optic nerves (ONs) of mice euthanized 6 MPI were processed for light and electron microscopy. Expression of mtHSP70Flag in the retina and ONs was evaluated by RT-PCR, immunofluorescence, and Western blotting. The activities of respiratory complexes I and III, as well as mitochondrial protein import were quantitated.
  
  
  RESULTS
  Expression: immunofluorescence revealed punctate and perinuclear expression of mtHSP70Flag that colocalized with mitochondrial porin in thy1.2 labeled retinal ganglion cells (RGCs). Immunoblotting and RT-PCR confirmed mtHSP70Flag expression in the retina and ON. Rescue: treatment with mtHSP70Flag resulted in a 44% increase in PERG amplitude and less delays in latency relative to the EAE-mCherry group that also showed progressive inner retinal thinning. At 6 MPI, the almost 50% loss of RGCs and optic nerve axons in EAE mice was suppressed by mtHSP70Flag. In addition, retinas of EAE-mtHSP70Flag mice showed nearly complete rescue of complex I and III activities that was reduced by one-third in the EAE-mCherry retinas. Lastly, reductions in import of COX8-mCherry into mitochondria of mice sensitized for EAE improved by 30% with mtHSP70Flag gene therapy.
  
  
  CONCLUSIONS
  Mitochondrial HSP70 ameliorates mitochondrial dysfunction that culminates in irreversible visual loss and atrophy of the optic nerve in EAE suggesting that it may be useful to prevent irreversible disability in patients with optic neuritis and multiple sclerosis (MS).
authors.affiliation: Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami, Miller School of Medicine, Miami, Florida, United States.
authors.forename: Venu
authors.initials: V
authors.lastname: Talla
authors.affiliation: Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami, Miller School of Medicine, Miami, Florida, United States.
authors.forename: Vittorio
authors.initials: V
authors.lastname: Porciatti
authors.affiliation: Department of Ophthalmology, University of Florida, Gainesville, Florida, United States.
authors.forename: Vince
authors.initials: V
authors.lastname: Chiodo
authors.affiliation: Department of Ophthalmology, University of Florida, Gainesville, Florida, United States.
authors.forename: Sanford L
authors.initials: SL
authors.lastname: Boye
authors.affiliation: Department of Ophthalmology, University of Florida, Gainesville, Florida, United States.
authors.forename: William W
authors.initials: WW
authors.lastname: Hauswirth
authors.affiliation: Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami, Miller School of Medicine, Miami, Florida, United States.
authors.forename: John
authors.initials: J
authors.lastname: Guy
mesh_terms: D000818:Animals; D001766:Blindness / Q000209:etiology / Q000628:therapy*; D004596:Electroretinography; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000150:complications* / Q000235:genetics; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000502:physiology / Q000627:therapeutic use*; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000378:metabolism*; D009896:Optic Atrophy / Q000209:etiology / Q000628:therapy*; D009900:Optic Nerve / Q000378:metabolism; D012160:Retina / Q000378:metabolism / Q000502:physiology; D012165:Retinal Ganglion Cells / Q000378:metabolism
chemical_list: D018840:HSP70 Heat-Shock Proteins; D024101:Mitochondrial Proteins
keywords: gene therapy; mtHSP70; optic neuropathy