Abstract
Abstract: :
Purpose:Currently a phase I clinical trial of adenoviral vector is now underway for intraocular neovascularization. We have previously reported that eyes with laser photocoagulation (PC) and photodynamic therapy increased and sustained transduction in the area of the treatment, which are already established for the treatment of retinal and/or choroidal neovascularization (Anzai K, et al. ARVO 2003). This study is designed to demonstrate the mechanisms of enhanced gene transduction by adenoviral vector in the PC treated retina. Methods:Five C57/BL6 mice received no treatment or fundus PC with 532 nm diode laser. Twenty–four hours after the treatment eyes were enucleated and total RNA was extracted from laser photocoagulated retina and choroid. Samples were processed for qRT–PCR (Prism 7700, Applied Biosystems) to quantify mRNA expression of coxsackie adenovirus receptor (CAR) and integrins αV, ß3 and ß5, which have been shown to play a role in cell surface interaction and internalization of viral vector during the transduction process. Results:Relative expression of integrin ß3 of retina with PC, retina without PC, choroid with PC, and choroid without PC were 2.08±0.31, 1.34±0.35, 2.11±0.27, and 1.41±0.42. The integrin ß3 mRNA expression was significantly upregulated in PC treated retina and choroid. Relative expression of integrin ß5 was also upregulated in the laser–treated retina (2.66±0.90) or choroid (0.38±0.19) when compared to the untreated retina (1.01±0.09) or choroid (0.20±0.12). There was no significant difference in relative expression of CAR between the eyes with or without PC treatment. Conclusions:In the laser treated retina and choroid, cell adhesion molecule; integrins are upregulated when compared to untreated retina and choroid. This, at least in part, may support our previous data demonstrating enhanced and sustained gene transduction by adenoviral vector in the PC treated retina.
Keywords: cell adhesions/cell junctions • laser • adenovirus