Direct in vivo gene transfer to urological organs

Purpose: Patients with urological disorders may benefit from gene based the rapy. We investigated the feasibility of delivering exogenous genes into ur ological tissues in vivo using direct in vivo electrotransfection. Materials and Methods: Gene transfer to rat kidneys, testes and bladders wa s accomplished via direct local injection of pGL3/luciferase and beta-galac tosidase reporter gene constructs, followed by an electrical pulse ranging from 55 to 115 msec. at 100 V. Direct injection of deoxyribonucleic acid wi thout an electrical pulse served as the control. The transfected and nontra nsfected organs were retrieved and analyzed by luciferase activity assay, h istochemical and immunocytochemical staining for beta-galactosidase, and re verse transcription polymerase chain reaction with primers specific for bet a-galactosidase messenger ribonucleic acid. Results: There was significant luciferase activity 1, 3 and 5 days after di rect in vivo electrotransfection in kidneys and testes, and after 3, 5, 7 a nd 10 days in bladders. Positive beta-galactosidase enzyme activity and bet a-galactosidase immunoreactivity were observed in the transfected renal tub ular cells, testicular interstitial and germ cells, and uroepithelial bladd er layer. Reverse transcription-polymerase chain reaction products of the t ransfected organs were noted, indicating the successful transcription of me ssenger ribonucleic acid. Conclusions: This study demonstrates that direct in vivo electrotransfectio n is a feasible method of transient gene delivery into intact urological or gans. Its apparent safety and relative simplicity suggest that direct in vi vo electrotransfection may be useful clinically.