PMID- 16708079 OWN - NLM STAT- MEDLINE DCOM- 20070412 LR - 20121115 IS - 0969-7128 (Print) IS - 0969-7128 (Linking) VI - 13 IP - 18 DP - 2006 Sep TI - AAV-mediated expression of CNTF promotes long-term survival and regeneration of adult rat retinal ganglion cells. PG - 1328-41 AB - 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. FAU - Leaver, S G AU - Leaver SG AD - School of Anatomy and Human Biology, The University of Western Australia, Western Australia, Australia. FAU - Cui, Q AU - Cui Q FAU - Plant, G W AU - Plant GW FAU - Arulpragasam, A AU - Arulpragasam A FAU - Hisheh, S AU - Hisheh S FAU - Verhaagen, J AU - Verhaagen J FAU - Harvey, A R AU - Harvey AR LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20060518 PL - England TA - Gene Ther JT - Gene therapy JID - 9421525 RN - 0 (Ciliary Neurotrophic Factor) RN - 147336-22-9 (Green Fluorescent Proteins) SB - IM MH - Animals MH - Axotomy MH - Cell Survival MH - Ciliary Neurotrophic Factor/analysis/*genetics/metabolism MH - Dependovirus/*genetics MH - Female MH - Gene Expression MH - Genetic Therapy/*methods MH - Genetic Vectors/*administration & dosage/genetics MH - Green Fluorescent Proteins/analysis/genetics MH - Immunohistochemistry MH - Injections MH - Nerve Regeneration MH - Optic Nerve Injuries/metabolism/pathology/*therapy MH - Rats MH - Rats, Wistar MH - Retinal Ganglion Cells/metabolism/pathology/virology MH - Reverse Transcriptase Polymerase Chain Reaction MH - Transduction, Genetic/*methods MH - Vitreous Body EDAT- 2006/05/19 09:00 MHDA- 2007/04/14 09:00 CRDT- 2006/05/19 09:00 PHST- 2006/05/19 09:00 [pubmed] PHST- 2007/04/14 09:00 [medline] PHST- 2006/05/19 09:00 [entrez] AID - 3302791 [pii] AID - 10.1038/sj.gt.3302791 [doi] PST - ppublish SO - Gene Ther. 2006 Sep;13(18):1328-41. doi: 10.1038/sj.gt.3302791. Epub 2006 May 18.