corneal transduction by intra-stromal injection of aav vectors in vivo in the mouse and ex vivo in human explants角膜转导intra-stromal注入aav载体体内的老鼠和人类外植体体外.pdfVIP

Corneal Transduction by Intra-Stromal Injection of AAV Vectors In Vivo in the Mouse and Ex Vivo in Human Explants Claire Hippert1,2.¤a, Sandy Ibanes1,2., Nicolas Serratrice1,2, Franck Court1,2, Franc¸ois Malecaze3,4, 1,2 1,2 ¤b Eric J. Kremer , Vasiliki Kalatzis * ´ ´ ´ ´ 1 Institut de Genetique Moleculaire de Montpellier, CNRS, Montpellier, France, 2 Universites Montpellier I II, Montpellier, France, 3 Inserm U563, Toulouse, France, ´ ˆ 4 Departement d’Ophtalmologie, Hopital Purpan, Toulouse, France Abstract The cornea is a transparent, avascular tissue that acts as the major refractive surface of the eye. Corneal transparency, assured by the inner stroma, is vital for this role. Disruption in stromal transparency can occur in some inherited or acquired diseases. As a consequence, light entering the eye is blocked or distorted, leading to decreased visual acuity. Possible treatment for restoring transparency could be via viral-based gene therapy. The stroma is particularly amenable to this strategy due to its immunoprivileged nature and low turnover rate. We assayed the potential of AAV vectors to transduce keratocytes following intra-stromal injection in vivo in the mouse cornea and ex vivo in human explants. In murine and human corneas, we transduced the entire stroma using a single injection, preferentially targeted keratocytes and achieved long-term gene transfer (up to 17 months in vivo in mice). Of the serotypes tested, AAV2/8 was the most promising for gene transfer in both mouse and man. Furthermore, transgene expression could be transiently increased following aggression to the cornea