title: Tyrosine triple mutated AAV2-BDNF gene therapy in a rat model of transient IOP elevation.
abstract: We examined the neuroprotective effects of exogenous brain-derived neurotrophic factor (BDNF), which provides protection to retinal ganglion cells (RGCs) in rodents, in a model of transient intraocular pressure (IOP) elevation using a mutant (triple Y-F) self-complementary adeno-associated virus type 2 vector encoding BDNF (tm-scAAV2-BDNF).
  
  
  
  The tm-scAAV2-BDNF or control vector encoding green fluorescent protein (GFP; tm-scAAV2-GFP) was intravitreally administered to rats, which were then divided into four groups: control, ischemia/reperfusion (I/R) injury only, I/R injury with tm-scAAV2-GFP, and tm-scAAV2-BDNF. I/R injury was then induced by transiently increasing IOP, after which the rats were euthanized to measure the inner retinal thickness and cell counts in the RGC layer.
  
  
  
  Intravitreous injection of tm-scAAV2-BDNF resulted in high levels of BDNF expression in the neural retina. Histological analysis showed that the inner retinal thickness and cell numbers in the RGC layer were preserved after transient IOP elevation in eyes treated with tm-scAAV2-BDNF but not in the other I/R groups. Significantly reduced glial fibrillary acidic protein (GFAP) immunostaining after I/R injury in the rats that received tm-scAAV2-BDNF indicated reduced retinal stress, and electroretinogram (ERG) analysis confirmed preservation of retinal function in the tm-scAAV2-BDNF group.
  
  
  
  These results demonstrate the feasibility and effectiveness of neuroprotective gene therapy using tm-scAAV2-BDNF to protect the inner retina from transiently high intraocular pressure. An in vivo gene therapeutic approach to the clinical management of retinal diseases in conditions such as glaucoma, retinal artery occlusion, hypertensive retinopathy, and diabetic retinopathy thus appears feasible.
authors.affiliation: Department of Ophthalmology, Nippon Medical School, Tokyo, Japan; Department of Biochemistry and Molecular Biology, Division of Gene Therapy Research Center for Advanced Medical Technology, Nippon Medical School, Tokyo, Japan.
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Igarashi
authors.affiliation: Department of Biochemistry and Molecular Biology, Division of Gene Therapy Research Center for Advanced Medical Technology, Nippon Medical School, Tokyo, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Miyake
authors.affiliation: Department of Ophthalmology, Nippon Medical School, Tokyo, Japan; Department of Biochemistry and Molecular Biology, Division of Gene Therapy Research Center for Advanced Medical Technology, Nippon Medical School, Tokyo, Japan.
authors.forename: Maika
authors.initials: M
authors.lastname: Kobayashi
authors.affiliation: Department of Ophthalmology, Nippon Medical School Chiba Hokusoh Hospital, Chiba, Japan.
authors.forename: Shuhei
authors.initials: S
authors.lastname: Kameya
authors.affiliation: Department of Ophthalmology, Nippon Medical School, Tokyo, Japan.
authors.forename: Chiaki
authors.initials: C
authors.lastname: Fujimoto
authors.affiliation: Department of Ophthalmology, Nippon Medical School, Tokyo, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Nakamoto
authors.affiliation: Department of Ophthalmology, Nippon Medical School, Tokyo, Japan.
authors.forename: Hisatomo
authors.initials: H
authors.lastname: Takahashi
authors.affiliation: Department of Pediatrics, Nippon Medical School, Tokyo, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Igarashi
authors.affiliation: Department of Biochemistry and Molecular Biology, Division of Gene Therapy Research Center for Advanced Medical Technology, Nippon Medical School, Tokyo, Japan.
authors.forename: Noriko
authors.initials: N
authors.lastname: Miyake
authors.affiliation: Department of Biochemistry and Molecular Biology, Division of Gene Therapy Research Center for Advanced Medical Technology, Nippon Medical School, Tokyo, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Iijima
authors.affiliation: Department of Biochemistry and Molecular Biology, Division of Gene Therapy Research Center for Advanced Medical Technology, Nippon Medical School, Tokyo, Japan.
authors.forename: Yukihiko
authors.initials: Y
authors.lastname: Hirai
authors.affiliation: Department of Biochemistry and Molecular Biology, Division of Gene Therapy Research Center for Advanced Medical Technology, Nippon Medical School, Tokyo, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimada
authors.affiliation: Department of Biochemistry and Molecular Biology, Division of Gene Therapy Research Center for Advanced Medical Technology, Nippon Medical School, Tokyo, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Okada
authors.affiliation: Department of Ophthalmology, Nippon Medical School, Tokyo, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Takahashi
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics* / Q000627:therapeutic use*; D002452:Cell Count; D000229:Dependovirus / Q000378:metabolism*; D004195:Disease Models, Animal; D004596:Electroretinography; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007429:Intraocular Pressure; D009154:Mutation / Q000235:genetics*; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000293:injuries / Q000473:pathology; D012165:Retinal Ganglion Cells / Q000378:metabolism / Q000473:pathology; D014161:Transduction, Genetic; D014443:Tyrosine / Q000235:genetics*
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D005904:Glial Fibrillary Acidic Protein; D049452:Green Fluorescent Proteins; D014443:Tyrosine