PMID- 19626133
OWN - NLM
STAT- MEDLINE
DCOM- 20091027
LR  - 20240104
IS  - 1090-0535 (Electronic)
IS  - 1090-0535 (Linking)
VI  - 15
DP  - 2009 Jul 17
TI  - Adeno-associated virus serotype-9 efficiently transduces the retinal outer 
      plexiform layer.
PG  - 1374-82
AB  - PURPOSE: Adeno-associated virus serotype-9 (AAV-9) is a promising gene delivery 
      vector. In this study, we evaluated AAV-9 transduction in the mouse retina. 
      METHODS: Three different AAV vectors were used in our study: AAV-9.RSV.AP, 
      AAV-9.CMV.eGFP, and AAV-9.CMV.R4-23/C. In these vectors, two different promoters 
      (the cytomegalovirus promoter-CMV promoter and the Rous sarcoma virus-RSV 
      promoter) were used to express three different transgenes including the alkaline 
      phosphatase (AP) gene, the enhanced green fluorescent protein (eGFP) gene, and a 
      therapeutic microdystrophin gene (the R4-23/C). Specifically, 1 microl AAV-9 
      reporter gene vectors (1x10(9) viral genome particles of AAV-9.RSV.AP or 1x10(10) 
      viral genome particles of AAV-9.CMV.eGFP) were administered subretinally to young 
      (2-3-week-old), adult (3-month-old), and old (12-month-old) C57BL/6J mice. To 
      evaluate AAV-9 transduction in a diseased retina, we injected subretinally 
      1x10(9) viral genome particles of AAV-9.CMV.R4-23/C to mdx(3cv) mice, which we 
      used as a model for Duchenne muscular dystrophy (DMD). Transgene expression was 
      examined by histochemical as well as immunofluorescence staining at three and 
      five weeks after injection. Electroretinograms were recorded five weeks after 
      subretinal AAV-9.RSV.AP injection. RESULTS: Subretinal injection yielded 
      widespread transduction throughout the retina in all age groups. Robust 
      expression was seen in the retinal pigment epithelium, outer nuclear layer, and 
      in Muller cells. Interestingly a synaptic layer, the outer plexiform layer (OPL), 
      also showed intensive expression. Transduction of the synaptic layer was further 
      confirmed by immunostaining for C-terminal binding protein 2 (CtBP2), a marker 
      for the photoreceptor synaptic ribbon. Dystrophin is normally expressed in the 
      OPL photoreceptor terminals. This expression is lost in DMD patients and mdx(3cv) 
      mice. Consistent with our findings in normal mice, we observed efficient 
      microdystrophin expression in the OPL after AAV-9.CMV.R4-23/C infection. At five 
      weeks after subretinal delivery of AAV-9.RSV.AP, no morphology or ERG 
      abnormalities were observed. CONCLUSIONS: We demonstrated that AAV-9 is a potent 
      vector for retinal gene delivery. Furthermore, subretinal AAV-9 administration 
      did not cause appreciable acute retinal damages. In summary, AAV-9-mediated OPL 
      transduction holds promise for treating diseases that primarily affect this 
      layer.
FAU - Lei, Bo
AU  - Lei B
AD  - Department of Ophthalmology, The First Affiliated Hospital of Chongqing Medical 
      University, 1 You Yi Road, Yu Zhong District, Chongqing, China. bolei99@126.com
FAU - Zhang, Keqing
AU  - Zhang K
FAU - Yue, Yongping
AU  - Yue Y
FAU - Ghosh, Arkasubhra
AU  - Ghosh A
FAU - Duan, Dongsheng
AU  - Duan D
LA  - eng
GR  - R01 AR049419/AR/NIAMS NIH HHS/United States
GR  - AR-49419/AR/NIAMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20090717
PL  - United States
TA  - Mol Vis
JT  - Molecular vision
JID - 9605351
RN  - 0 (Dystrophin)
RN  - 0 (enhanced green fluorescent protein)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Darkness
MH  - Dependovirus/*classification
MH  - Dystrophin/metabolism
MH  - Electroretinography
MH  - Green Fluorescent Proteins/metabolism
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Retina/*cytology/metabolism/*virology
MH  - Serotyping
MH  - *Transduction, Genetic
PMC - PMC2713732
EDAT- 2009/07/25 09:00
MHDA- 2009/10/29 06:00
PMCR- 2009/01/01
CRDT- 2009/07/24 09:00
PHST- 2009/02/10 00:00 [received]
PHST- 2009/06/21 00:00 [accepted]
PHST- 2009/07/24 09:00 [entrez]
PHST- 2009/07/25 09:00 [pubmed]
PHST- 2009/10/29 06:00 [medline]
PHST- 2009/01/01 00:00 [pmc-release]
AID - 145 [pii]
AID - 2009molvis0039 [pii]
PST - epublish
SO  - Mol Vis. 2009 Jul 17;15:1374-82.