Stromelysin gene transfer into cultured human trabecular cells and rat trabecular meshwork in vivo

PURPOSE. To determine whether stromelysin gene can be introduced into and e xpressed in the cultured human trabecular cells as well as in the rat eye i n vivo through means of a recombinant replication-deficient adenovirus. METHODS. Stromelysin cDNA was obtained by reverse transcription-polymerase chain reaction with mRNA extracted from the cultured human trabecular cells after induction with interleukin 1 alpha. Adenovirus vector that contains stromelysin cDNA was constructed by cotransfection of pJM17 and p DeltaA. C MV-str into the 293 cells. The expression of stromelysin in the cultured hu man trabecular cells was assayed by Western blot and zymography. The expres sion of stromelysin in the trabecular meshwork of the rat eyes was detected by in situ hybridization and immunohistochemistry. RESULTS. The constructed adenovirus vector contained Stromelysin cDNA, but no E1 region. Western blot and zymogram revealed that the stromelysin could be expressed and that it possessed enzymatic activity in cultured human tr abecular cells. In situ hybridization and immunostaining of the Stromelysin showed that the complete form of stromelysin was expressed in the trabecul ar meshwork, the iris, and the uveoscleral outflow pathway of the rat eye. CONCLUSIONS. Stromelysin, a functional gene, can be transferred in vivo int o rat eyes and in vitro into cultured human trabecular cells using a replic ation-deficient adenovirus vector. This shows the possibility of gene thera py in glaucoma.