Alternate serotype adenovector provides long-term therapeutic gene expression in the eye

Mol Vis. 2008;14:2535-46. Epub 2008 Dec 30.

Abstract

Purpose: To determine whether the duration of transgene expression from an alternate adenovector serotype, Ad35, can provide advantages over an Ad5 serotype vector following a single intravitreal (IVT) administration.

Methods: To assess the transgene expression profile, mice received one IVT injection of Ad5- or Ad35-based vectors expressing green fluorescent protein (GFP), luciferase or pigment epithelium-derived factor (PEDF). At specified time points following vector administration, eyes were monitored for GFP expression, or eyes were harvested and assayed for adenovector genomes, luciferase activity or PEDF levels. Ad35-based vector in vivo biologic activity was investigated using a mouse model of laser-induced choroidal neovascularization (CNV). On Day 0, mice received one IVT injection of Ad5.PEDF or Ad35.PEDF (HI-RGD) followed by laser-induced CNV on Day 28. Fourteen days later, animals were perfused with fluorescein-labeled dextran and CNV lesion size quantitated in choroidal flat mounts.

Results: These studies demonstrate that following a single IVT adenovector administration: 1) gene expression is prolonged following administration of an Ad35 compared to an Ad5-based vector; 2) the amount of vector genomes in the eye remain constant out to 60 days post injection of both Ad5 and Ad35-based vectors; and 3) an Ad35.PEDF (HI-RGD) vector inhibits CNV in a mouse model at 42 days post injection.

Conclusions: These studies show that transgene and genome levels are prolonged in the eye following 1 IVT injection of an Ad35-based vector. Moreover, therapeutic gene levels from 1 IVT administration of Ad35.PEDF (HI-RGD) vector block abnormal blood vessel growth in a laser-induced CNV mouse model.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adenoviridae / classification*
  • Adenoviridae / genetics*
  • Animals
  • Choroidal Neovascularization / genetics
  • Choroidal Neovascularization / pathology
  • Choroidal Neovascularization / therapy
  • Eye / metabolism*
  • Eye / pathology
  • Eye Proteins / genetics
  • Eye Proteins / therapeutic use
  • Female
  • Genetic Therapy / methods*
  • Genetic Vectors / genetics*
  • Genome
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Injections
  • Luciferases / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Nerve Growth Factors / genetics
  • Nerve Growth Factors / therapeutic use
  • Polymerase Chain Reaction
  • Serotyping
  • Serpins / genetics
  • Serpins / therapeutic use
  • Time Factors
  • Transgenes

Substances

  • Eye Proteins
  • Nerve Growth Factors
  • Serpins
  • pigment epithelium-derived factor
  • Green Fluorescent Proteins
  • Luciferases