Photoreceptor protection by adeno-associated virus-mediated LEDGF expression in the RCS rat model of retinal degeneration: probing the mechanism

Invest Ophthalmol Vis Sci. 2009 Aug;50(8):3897-906. doi: 10.1167/iovs.08-3153. Epub 2009 Mar 25.

Abstract

Purpose: Lens epithelium-derived growth factor (LEDGF) is upregulated in response to stress and enhances the survival of neurons in the retina and optic nerve, as well as a wide range of other cells, such as fibroblasts and keratinocytes. Photoreceptor protection was investigated in the RCS rat retinal degeneration model after Ledgf delivery with an adeno-associated virus (AAV) and the mechanism of protection explored.

Methods: Thirty-six RCS and nine P23H rats had bilateral subretinal injections of AAV-Ledgf in one eye and buffer in the contralateral eye as the control. Retinal function was evaluated 8 weeks later by the electroretinogram and compared with photoreceptor cell layer count. LEDGF mRNA and protein levels and mRNA levels of known stress-related factors were compared in treated and control retinas to explore the mechanism of LEDGF protection. Nine RCS rats were treated with adenovirus-heat shock protein 27 (Ad-HSP27) and examined for protection.

Results: Significant photoreceptor protection was evident functionally and morphologically in 65% to 100% of the RCS rats treated at early ages of up to 7 weeks. Cell protection was more prominent in the superior retinal hemisphere which has a slower natural degeneration rate in untreated eyes. Although many of the heat shock proteins and other stress-related genes showed significant elevation in the AAV-Ledgf-treated eyes, all increases were approximately twofold or less. Transduction of retinal cells with Ad-HSP27 also resulted in photoreceptor protection. AAV-Ledgf elicited no photoreceptor functional protection in P23H rhodopsin transgenic rat retina.

Conclusions: Chronic LEDGF treatment via AAV-Ledgf administration gave successful protection of photoreceptors in the RCS rat retina and retarded cell death by about 2 weeks. Induction of heat shock proteins also gave photoreceptor protection. However, compelling evidence was not found that LEDGF protection was associated with upregulation of heat shock proteins.

Publication types

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

MeSH terms

  • Adaptor Proteins, Signal Transducing / genetics*
  • Adaptor Proteins, Signal Transducing / metabolism
  • Animals
  • Animals, Genetically Modified
  • Blotting, Western
  • Cell Count
  • Cell Survival
  • Cytoprotection
  • Dependovirus / genetics*
  • Electroretinography
  • Gene Expression Regulation / physiology*
  • Genetic Therapy / methods*
  • Genetic Vectors
  • HSP27 Heat-Shock Proteins / genetics
  • HSP27 Heat-Shock Proteins / metabolism
  • Mutagenesis, Site-Directed
  • Photoreceptor Cells, Vertebrate / physiology*
  • Plasmids
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Mutant Strains
  • Retinal Degeneration / metabolism
  • Retinal Degeneration / physiopathology
  • Retinal Degeneration / therapy*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Transduction, Genetic
  • Up-Regulation

Substances

  • Adaptor Proteins, Signal Transducing
  • HSP27 Heat-Shock Proteins
  • Psip1 protein, rat
  • RNA, Messenger
  • Transcription Factors