PMID- 27439313
OWN - NLM
STAT- MEDLINE
DCOM- 20170823
LR  - 20220409
IS  - 1557-7422 (Electronic)
IS  - 1043-0342 (Print)
IS  - 1043-0342 (Linking)
VI  - 27
IP  - 8
DP  - 2016 Aug
TI  - Highly Efficient Delivery of Adeno-Associated Viral Vectors to the Primate 
      Retina.
PG  - 580-97
LID - 10.1089/hum.2016.085 [doi]
AB  - Adeno-associated virus (AAV) has emerged as the preferred vector for targeting 
      gene expression to the retina. Subretinally injected AAV can efficiently 
      transduce retinal pigment epithelium and photoreceptors in primate retina. Inner 
      and middle primate retina can be transduced by intravitreally delivered AAV, but 
      with low efficiency. This is due to dilution of vector, potential neutralization 
      of capsid because it is not confined to the immune-privileged retinal 
      compartment, and the presence of the inner limiting membrane (ILM), a barrier 
      separating the vitreous from the neural retina. We here describe a novel "subILM" 
      injection method that addresses all three issues. Specifically, vector is placed 
      in a surgically induced, hydrodissected space between the ILM and neural retina. 
      In an initial experiment, we injected viscoelastic (Healon((R))), a substance we 
      confirmed was biocompatible with AAV, to create a subILM bleb and subsequently 
      injected AAV2-GFP into the bleb after irrigation with basic salt solution. For 
      later experiments, we used a Healon-AAV mixture to place single, subILM 
      injections. In all cases, subILM delivery of AAV was well tolerated-no 
      inflammation or gross structural changes were observed by ophthalmological 
      examination or optical coherence tomography. In-life fluorescence imaging 
      revealed profound transgene expression within the area of the subILM injection 
      bleb that persisted for the study duration. Uniform and extensive transduction of 
      retinal ganglion cells (RGCs) was achieved in the areas beneath the subILM bleb. 
      Transduction of Muller glia, ON bipolar cells, and photoreceptors was also 
      observed. Robust central labeling from green fluorescent protein-expressing RGCs 
      confirmed their continued survival, and was observed in the lateral geniculate 
      nucleus, the superior colliculus, and the pretectum. Our results confirm that the 
      ILM is a major barrier to transduction by AAV in primate retina and that, when it 
      is circumvented, the efficiency and depth to which AAV2 promotes transduction of 
      multiple retinal cell classes are greatly enhanced.
FAU - Boye, Shannon E
AU  - Boye SE
AD  - 1 Department of Ophthalmology, University of Florida College of Medicine , 
      Gainesville, Florida.
FAU - Alexander, John J
AU  - Alexander JJ
AD  - 2 Department of Human Genetics, Emory University , Atlanta, Georgia.
FAU - Witherspoon, C Douglas
AU  - Witherspoon CD
AD  - 3 Department of Ophthalmology, University of Alabama at Birmingham , Birmingham, 
      Alabama.
FAU - Boye, Sanford L
AU  - Boye SL
AD  - 1 Department of Ophthalmology, University of Florida College of Medicine , 
      Gainesville, Florida.
FAU - Peterson, James J
AU  - Peterson JJ
AD  - 1 Department of Ophthalmology, University of Florida College of Medicine , 
      Gainesville, Florida.
FAU - Clark, Mark E
AU  - Clark ME
AD  - 3 Department of Ophthalmology, University of Alabama at Birmingham , Birmingham, 
      Alabama.
FAU - Sandefer, Kristen J
AU  - Sandefer KJ
AD  - 4 Department of Neurology, University of Alabama at Birmingham , Birmingham, 
      Alabama.
FAU - Girkin, Chris A
AU  - Girkin CA
AD  - 3 Department of Ophthalmology, University of Alabama at Birmingham , Birmingham, 
      Alabama.
FAU - Hauswirth, William W
AU  - Hauswirth WW
AD  - 1 Department of Ophthalmology, University of Florida College of Medicine , 
      Gainesville, Florida.
FAU - Gamlin, Paul D
AU  - Gamlin PD
AD  - 3 Department of Ophthalmology, University of Alabama at Birmingham , Birmingham, 
      Alabama.
LA  - eng
GR  - R01 EY024280/EY/NEI NIH HHS/United States
GR  - R01 EY025555/EY/NEI NIH HHS/United States
GR  - P30 EY021721/EY/NEI NIH HHS/United States
GR  - P30 EY003039/EY/NEI NIH HHS/United States
GR  - UL1 RR025777/RR/NCRR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Hum Gene Ther
JT  - Human gene therapy
JID - 9008950
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Dependovirus/*genetics
MH  - Genetic Vectors/*administration & dosage
MH  - Green Fluorescent Proteins/metabolism
MH  - Macaca nemestrina
MH  - Male
MH  - Retina/*metabolism/pathology
MH  - Retinal Ganglion Cells/*metabolism/pathology
MH  - Transduction, Genetic
MH  - Transgenes/*genetics
PMC - PMC4991591
EDAT- 2016/07/22 06:00
MHDA- 2017/08/24 06:00
PMCR- 2017/08/01
CRDT- 2016/07/22 06:00
PHST- 2016/07/22 06:00 [entrez]
PHST- 2016/07/22 06:00 [pubmed]
PHST- 2017/08/24 06:00 [medline]
PHST- 2017/08/01 00:00 [pmc-release]
AID - 10.1089/hum.2016.085 [pii]
AID - 10.1089/hum.2016.085 [doi]
PST - ppublish
SO  - Hum Gene Ther. 2016 Aug;27(8):580-97. doi: 10.1089/hum.2016.085.