TY - JOUR
T1 - AAV-mediated expression of CNTF promotes long-term survival and regeneration of adult rat retinal ganglion cells
AU - Leaver, Simone
AU - Cui, Q.
AU - Plant, Giles
AU - Arulpragasam, Ajanthy
AU - Hisheh, S.
AU - Verhaagen, J.
AU - Harvey, Alan
PY - 2006
Y1 - 2006
N2 - We compared the effects of intravitreal injection of bicistronic 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 beta III-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: 17450 +/- 358 s.e.m.) and AAV-BDNF-GFP injected eyes (10200 +/- 4064 RGCs/retina). Consistent with increased RGC viability in AAV-CNTF-GFP and AAV-BDNF-GFP injected eyes, these animals possessed many beta III-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 ( 11357367 axons/nerve, 0.5mm 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 ( 72833) surviving RGCs/retina, about 25% of the adult RGC population. Of these, 13 352 (71868) 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.
AB - We compared the effects of intravitreal injection of bicistronic 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 beta III-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: 17450 +/- 358 s.e.m.) and AAV-BDNF-GFP injected eyes (10200 +/- 4064 RGCs/retina). Consistent with increased RGC viability in AAV-CNTF-GFP and AAV-BDNF-GFP injected eyes, these animals possessed many beta III-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 ( 11357367 axons/nerve, 0.5mm 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 ( 72833) surviving RGCs/retina, about 25% of the adult RGC population. Of these, 13 352 (71868) 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.
U2 - 10.1038/sj.gt.3302791
DO - 10.1038/sj.gt.3302791
M3 - Article
C2 - 16708079
SN - 0969-7128
VL - 13
SP - 1328
EP - 1341
JO - Gene Therapy
JF - Gene Therapy
ER -