title: Ocular localization and transduction by adenoviral vectors are serotype-dependent and can be modified by inclusion of RGD fiber modifications.
abstract: OBJECTIVE
  To evaluate localization and transgene expression from adenoviral vector of serotypes 5, 35, and 28, ± an RGD motif in the fiber following intravitreal or subretinal administration.
  
  
  METHODS
  Ocular transduction by adenoviral vector serotypes ± RGD was studied in the eyes of mice receiving an intravitreous or subretinal injection. Each serotype expressed a CMV-GFP expression cassette and histological sections of eyes were examined. Transgene expression levels were examined using luciferase (Luc) regulated by the CMV promoter.
  
  
  RESULTS
  GFP localization studies revealed that serotypes 5 and 28 given intravitreously transduced corneal endothelial, trabecular, and iris cells. Intravitreous delivery of the unmodified Ad35 serotype transduced only trabecular meshwork cells, but, the modification of the RGD motif into the fiber of the Ad35 viral vector base expanded transduction to corneal endothelial and iris cells. Incorporation of the RGD motif into the fiber knob with deletion of RGD from the penton base did not affect the transduction ability of the Ad5 vector base. Subretinal studies showed that RGD in the Ad5 knob shifted transduction from RPE cells to photoreceptor cells. Using a CMV-Luc expression cassette, intravitreous delivery of all the tested vectors, such as Ad5-, Ad35- and Ad28- resulted in an initial rapid induction of luciferase activity that thereafter declined. Subretinal administration of vectors showed a marked difference in transgene activity. Ad35-Luc gene expression peaked at 7 days and remained elevated for 6 months. Ad28-Luc expression was high after 1 day and remained sustained for one month.
  
  
  CONCLUSIONS
  Different adenoviral vector serotypes ± modifications transduce different cells within the eye. Transgene expression can be brief or extended and is serotype and delivery route dependent. Thus, adenoviral vectors provide a versatile platform for the delivery of therapeutic agents for ocular diseases.
authors.affiliation: Department of Ophthalmology, Saitama Medical University, Moroyama, Iruma, Saitama, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Ueyama
authors.affiliation: Department of Ophthalmology, Saitama Medical University, Moroyama, Iruma, Saitama, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Mori
authors.affiliation: Department of Ophthalmology, Saitama Medical University, Moroyama, Iruma, Saitama, Japan.
authors.forename: Takuhei
authors.initials: T
authors.lastname: Shoji
authors.affiliation: Department of Ophthalmology, Saitama Medical University, Moroyama, Iruma, Saitama, Japan.
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Omata
authors.affiliation: Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Peter L
authors.initials: PL
authors.lastname: Gehlbach
authors.affiliation: GenVec, Inc., Gaithersburg, Maryland, United States of America.
authors.forename: Douglas E
authors.initials: DE
authors.lastname: Brough
authors.affiliation: GenVec, Inc., Gaithersburg, Maryland, United States of America; National Eye Institute, Bethesda, Maryland, United States of America.
authors.forename: Lisa L
authors.initials: LL
authors.lastname: Wei
authors.affiliation: Department of Ophthalmology, Saitama Medical University, Moroyama, Iruma, Saitama, Japan.
authors.forename: Shin
authors.initials: S
authors.lastname: Yoneya
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D005260:Female; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D056965:Injections, Intraocular; D008810:Mice, Inbred C57BL; D009842:Oligopeptides / Q000235:genetics*; D014161:Transduction, Genetic
chemical_list: D009842:Oligopeptides; D049452:Green Fluorescent Proteins; C047981:arginyl-glycyl-aspartic acid