AAV-mediated expression of CNTF promotes long-term survival and regeneration of adult rat retinal ganglion cells

Simone G Leaver, Qi Cui, Giles W Plant, A. Arulpragasam, S Hisheh, J Verhaagen, Alan R Harvey

Research output: Contribution to journal/periodicalArticleScientificpeer-review

245 Citations (Scopus)

Abstract

We compared the effects of intravitreal injection of bi-cistronic adeno-associated viral (AAV-2) vectors encoding enhanced green fluorescent protein (GFP) and either ciliary neurotrophic factor (CNTF), brain-derived neurotrophic factor (BDNF) or growth-associated protein-43 (GAP43) on adult retinal ganglion cell (RGC) survival and regeneration following (i) optic nerve (ON) crush or (ii) after ON cut and attachment of a peripheral nerve (PN). At 7 weeks after ON crush, quantification of betaIII-tubulin immunostaining revealed that, compared to AAV-GFP controls, RGC survival was not enhanced by AAV-GAP43-GFP but was increased in AAV-CNTF-GFP (mean RGCs/retina: 17 450+/-358 s.e.m.) and AAV-BDNF-GFP injected eyes (10 200+/-4064 RGCs/retina). Consistent with increased RGC viability in AAV-CNTF-GFP and AAV-BDNF-GFP injected eyes, these animals possessed many betaIII-tubulin- and GFP-positive fibres proximal to the ON crush. However, only in the AAV-CNTF-GFP group were regenerating RGC axons seen in distal ON (1135+/-367 axons/nerve, 0.5 mm post-crush), some reaching the optic chiasm. RGCs were immunoreactive for CNTF and quantitative RT-PCR revealed a substantial increase in CNTF mRNA expression in retinas transduced with AAV-CNTF-GFP. The combination of AAV-CNTF-GFP transduction of RGCs with autologous PN-ON transplantation resulted in even greater RGC survival and regeneration. At 7 weeks after PN transplantation there were 27 954 (+/-2833) surviving RGCs/retina, about 25% of the adult RGC population. Of these, 13 352 (+/-1868) RGCs/retina were retrogradely labelled after fluorogold injections into PN grafts. In summary, AAV-mediated expression of CNTF promotes long-term survival and regeneration of injured adult RGCs, effects that are substantially enhanced by combining gene and cell-based therapies/interventions.

Original languageEnglish
Pages (from-to)1328-41
Number of pages14
JournalGene Therapy
Volume13
Issue number18
DOIs
Publication statusPublished - Sept 2006

Keywords

  • Animals
  • Axotomy
  • Cell Survival
  • Ciliary Neurotrophic Factor
  • Dependovirus
  • Female
  • Gene Expression
  • Genetic Therapy
  • Genetic Vectors
  • Green Fluorescent Proteins
  • Immunohistochemistry
  • Injections
  • Nerve Regeneration
  • Optic Nerve Injuries
  • Rats
  • Rats, Wistar
  • Retinal Ganglion Cells
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transduction, Genetic
  • Vitreous Body
  • Journal Article
  • Research Support, Non-U.S. Gov't

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