Adenovirus‐mediated gene transfer to murine retinal cells in vitro and in vivo

Abstract

Adenovirus-mediated gene transfer to retinal cells was evaluated using the replication-defective recombinant adenovirus vector Ad2/CMVlacZ-1 (coding for /I-galactosidase) both in an in vitro murine culture model and in vivo in adult mice. In vitro, no difference in infectability of neuronal and glial cells was observed, and 5096 of neurons expressed the exogenous gene at low viral concentration (10 pfulcell). In vivo, intraocular injection of 3 x IO6 pfu Ad2/CMVlacZ-1 resulted in expression of the transferred /3galactosidase gene in retinal pigment epithelium and ganglion cells. These results demonstrate that Ad2lCMVkzcZ-1 is an effective vector for gene transfer into retinal cells. &y wor& Defective recombinant adenovirus; /I-Galactosidase; Retina; Gene transfer; Retinal cell culture Introductioll

A vitrectomy improves the transfection efficiency of adenoviral vector-mediated gene transfer to Müller cells

“…3 An adenovirus-mediated gene transfection depends on the direct contact of the virus with the recipient cells. 18 The vitreous body can be a large obstacle to direct contact between an adenovirus and the retina.…”

“…[1][2][3][4][5][6][7][8][9][10][11][12][13][14] In common retinal diseases such as diabetic retinopathy, proliferative vitreoretinopathy and glaucoma, the neural retina is a logical target for the transfer of genes for therapeutic purposes, because the neural retina is primarily damaged in most of these diseases and various attempts have been made to either protect or increase the healing of the neural retina. [3][4][5]7,9,10 The adenoviral vector-mediated gene transfer to the retina can be classified into the following two groups, an extra-ocular injection or an intra-ocular injection.…”

“…[1][2][3][4][5][6][7][8][9][10][11][12][13][14] In common retinal diseases such as diabetic retinopathy, proliferative vitreoretinopathy and glaucoma, the neural retina is a logical target for the transfer of genes for therapeutic purposes, because the neural retina is primarily damaged in most of these diseases and various attempts have been made to either protect or increase the healing of the neural retina. [3][4][5]7,9,10 The adenoviral vector-mediated gene transfer to the retina can be classified into the following two groups, an extra-ocular injection or an intra-ocular injection. [1][2][3][4][5][6][7][8] Cayoutte et al 7 reported that the injection of the adenoviral vector into the superior colluculus can deliver genes to the retinal ganglion cells, however, the brain is also transfected by this procedure and as a result, the procedure itself (namely, an injection into the brain) might induce more serious problems in the patients.…”

“…[3][4][5]7,9,10 The adenoviral vector-mediated gene transfer to the retina can be classified into the following two groups, an extra-ocular injection or an intra-ocular injection. [1][2][3][4][5][6][7][8] Cayoutte et al 7 reported that the injection of the adenoviral vector into the superior colluculus can deliver genes to the retinal ganglion cells, however, the brain is also transfected by this procedure and as a result, the procedure itself (namely, an injection into the brain) might induce more serious problems in the patients. Regarding intra-ocular injections, a subretinal injection can deliver genes to the retina but is limited to the retinal pigment epithelium of adult animals and the subretinal injection procedure itself sometimes induces apoptosis of photoreceptor cells.…”

“…[1][2][3][4][5][6][7][8] This is because an adenoviral vector can transfer genes to nondividing cells with high efficiency. [1][2][3][4][5][6][7][8]11,12 In spite of this advantage, there are still several problems concerning the use of an adenoviral vector, one of which is the possibility of inducing an inflammation due to the expression of viral proteins which might induce irreversible damage to the retinal 1089 function. 11,12 There has yet to be a report which studies the function of the gene-transfected retina.…”