PMID- 21212183
OWN - NLM
STAT- MEDLINE
DCOM- 20110615
LR  - 20220224
IS  - 1552-5783 (Electronic)
IS  - 0146-0404 (Print)
IS  - 0146-0404 (Linking)
VI  - 52
IP  - 5
DP  - 2011 Apr
TI  - Whirlin replacement restores the formation of the USH2 protein complex in whirlin 
      knockout photoreceptors.
PG  - 2343-51
LID - 10.1167/iovs.10-6141 [doi]
AB  - PURPOSE: Whirlin is the causative gene for Usher syndrome type IID (USH2D), a 
      condition manifested as both retinitis pigmentosa and congenital deafness. 
      Mutations in this gene cause disruption of the USH2 protein complex composed of 
      USH2A and VLGR1 at the periciliary membrane complex (PMC) in photoreceptors. In 
      this study, the adeno-associated virus (AAV)-mediated whirlin replacement was 
      evaluated as a treatment option. METHODS: Murine whirlin cDNA driven by the human 
      rhodopsin kinase promoter (hRK) was packaged as an AAV2/5 vector and delivered 
      into the whirlin knockout retina through subretinal injection. The efficiency, 
      efficacy, and safety of this treatment were examined using immunofluorescent 
      staining, confocal imaging, immunoelectron microscopy, Western blot analysis, 
      histologic analysis, and electroretinogram. RESULTS: The AAV-mediated whirlin 
      expression started at two weeks, reached its maximum level at 10 weeks, and 
      lasted up to six months post injection. The transgenic whirlin product had a 
      molecular size and an expression level comparable to the wild-type. It was 
      distributed at the PMC in both rod and cone photoreceptors from the central to 
      peripheral retina. Importantly, the transgenic whirlin restored the cellular 
      localization and expression level of both USH2A and VLGR1 and did not cause 
      defects in the retinal histology and function in the whirlin knockout mouse. 
      CONCLUSIONS: Whirlin transgene recruits USH2A and VLGR1 to the PMC and is 
      sufficient for the formation of the USH2 protein complex in photoreceptors. The 
      combined hRK and AAV gene delivery system could be an effective gene therapy 
      approach to treat retinal degeneration in USH2D patients.
CI  - Copyright 2011 The Association for Research in Vision and Ophthalmology, Inc.
FAU - Zou, Junhuang
AU  - Zou J
AD  - Department of Ophthalmology and Visual Sciences, Moran Eye Center, University of 
      Utah, Salt Lake City, Utah 84132, USA.
FAU - Luo, Ling
AU  - Luo L
FAU - Shen, Zuolian
AU  - Shen Z
FAU - Chiodo, Vince A
AU  - Chiodo VA
FAU - Ambati, Balamurali K
AU  - Ambati BK
FAU - Hauswirth, William W
AU  - Hauswirth WW
FAU - Yang, Jun
AU  - Yang J
LA  - eng
GR  - R01 EY020853/EY/NEI NIH HHS/United States
GR  - R01 EY08571/EY/NEI NIH HHS/United States
GR  - P30 EY008571/EY/NEI NIH HHS/United States
GR  - P30 EY014800-07/EY/NEI NIH HHS/United States
GR  - P30 EY021721/EY/NEI NIH HHS/United States
GR  - P30 EY014800/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 - 20110412
PL  - United States
TA  - Invest Ophthalmol Vis Sci
JT  - Investigative ophthalmology & visual science
JID - 7703701
RN  - 0 (Extracellular Matrix Proteins)
RN  - 0 (Mass1 protein, mouse)
RN  - 0 (Membrane Proteins)
RN  - 0 (Receptors, G-Protein-Coupled)
RN  - 0 (Ush2a protein, mouse)
RN  - 0 (Whrn protein, mouse)
RN  - EC 2.7.11.14 (G-Protein-Coupled Receptor Kinase 1)
SB  - IM
MH  - Animals
MH  - Animals, Newborn
MH  - Blotting, Western
MH  - Dependovirus/genetics
MH  - Electroretinography
MH  - Extracellular Matrix Proteins/*genetics/metabolism
MH  - Fluorescent Antibody Technique, Indirect
MH  - G-Protein-Coupled Receptor Kinase 1/genetics
MH  - Gene Deletion
MH  - Gene Expression Regulation/*physiology
MH  - *Genetic Vectors
MH  - Membrane Proteins/*genetics/metabolism
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Mice, Knockout
MH  - Microscopy, Confocal
MH  - Microscopy, Immunoelectron
MH  - Photoreceptor Cells, Vertebrate/*metabolism
MH  - Plasmids
MH  - Polymerase Chain Reaction
MH  - Promoter Regions, Genetic
MH  - Receptors, G-Protein-Coupled/genetics/metabolism
MH  - Transgenes
MH  - Usher Syndromes/genetics
PMC - PMC3081228
EDAT- 2011/01/08 06:00
MHDA- 2011/06/16 06:00
PMCR- 2011/10/01
CRDT- 2011/01/08 06:00
PHST- 2011/01/08 06:00 [entrez]
PHST- 2011/01/08 06:00 [pubmed]
PHST- 2011/06/16 06:00 [medline]
PHST- 2011/10/01 00:00 [pmc-release]
AID - iovs.10-6141 [pii]
AID - 10-6141 [pii]
AID - 10.1167/iovs.10-6141 [doi]
PST - epublish
SO  - Invest Ophthalmol Vis Sci. 2011 Apr 12;52(5):2343-51. doi: 10.1167/iovs.10-6141. 
      Print 2011 Apr.