Surgical Removal of Internal Limiting Membrane and Layering of AAV Vector on the Retina Under Air Enhances Gene Transfection in a Nonhuman Primate

Investigative Ophthalmology & Visual Science
Kelvin Yi Chong TeoIan Jeffery Constable

Abstract

To determine if the surgical removal of the internal limiting membrane (ILM) in nonhuman primates (NHPs) will result in safe and effective transfection of adeno-associated viral (AAV2) vectors using green fluorescent protein (GFP) as a reporter. Six Macaca fascicularis NHP eyes underwent vitrectomy, ILM peel with layering of 1.7 × 1013 genome copies per milliliter of AAV2-GFP under air. Four control eyes underwent only vitrectomy and pooling under air. The intensity and area transfected was quantified in vivo with fundus autofluorescence (FAF) imaging. NHPs were euthanized 16 weeks postsurgery and immunohistochemical analysis assessed GFP expression at the cellular level. There was a larger area of fluorescence in ILM peeled eyes then in non-ILM peeled eyes (50.7 [33.1-58.4] pixel2 versus 5.1 [0.6-7.6] pixel2, P < 0.01). The intensity of fluorescence was also higher in ILM peeled eyes (10.3 [2.2-18.5] vs. 1.9 [0.6-4.4], P = 0.05). Non-ILM peeled eyes displayed fluorescence confined to the foveal center. Histological sections showed colocalization in the Müller cell layer, ganglion cell layer, and photoreceptor cell layer in the ILM peeled eyes. In non-ILM peeled eyes GFP expression was only in the ganglion cell layer in three e...Continue Reading

References

Feb 22, 2000·American Journal of Ophthalmology·N HagimuraS Kishi
Aug 6, 2000·American Journal of Ophthalmology·V Mester, F Kuhn
Jun 25, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Yu ZhouAdriana Di Polo
Jul 19, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Chooi-May LaiElizabeth P Rakoczy
Jul 22, 2005·Clinical & Experimental Ophthalmology·Alvin Kh KwokKenneth Sc Yuen
Mar 1, 2006·The Journal of Gene Medicine·Teresa BorrásSarah S Chisolm
Oct 6, 2006·The New England Journal of Medicine·Philip J RosenfeldUNKNOWN MARINA Study Group
Feb 23, 2007·International Journal of Clinical Practice·P K Kaiser, D V Do
Jul 3, 2007·American Journal of Ophthalmology·Ates YanyaliAhmet F Nohutcu
Aug 13, 2009·Molecular Therapy : the Journal of the American Society of Gene Therapy·Deniz DalkaraJohn G Flannery
May 29, 2010·Ophthalmic Surgery, Lasers & Imaging : the Official Journal of the International Society for Imaging in the Eye·Sachin MehtaM Gilbert Grand
Feb 12, 2011·Investigative Ophthalmology & Visual Science·Lu YinWilliam H Merigan
Jun 24, 2011·Science Translational Medicine·Luk H VandenbergheJean Bennett
Sep 14, 2011·Archives of Ophthalmology·T Michael NorkBrian J Christian
May 9, 2012·Lancet·Laurence S LimTien Y Wong
Feb 6, 2013·PloS One·Luk H VandenbergheJames M Wilson
Jan 24, 2015·European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik E.V·Christian SchönStylianos Michalakis
Dec 10, 2015·Investigative Ophthalmology & Visual Science·James BellinghamJacqueline van der Spuy
Apr 5, 2016·Clinical Ophthalmology·Kamyar VaziriHarry W Flynn
Jul 22, 2016·Human Gene Therapy·Shannon E BoyePaul D Gamlin
Jan 28, 2017·Molecular Therapy : the Journal of the American Society of Gene Therapy·Kazuhisa TakahashiTakashi Okada
Feb 6, 2017·Human Molecular Genetics·Jeffrey O'CallaghanPeter Humphries

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Citations

Jun 25, 2020·Human Gene Therapy·Sophia-Martha Kleine HolthausRobin R Ali
Mar 24, 2021·Experimental Eye Research·Kevin Y Zhang, Thomas V Johnson
Apr 18, 2021·Trends in Pharmacological Sciences·Cole W PetersKillian S Hanlon
Jun 4, 2021·Bioconjugate Chemistry·Marco BassettoOlivier Zelphati

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