Adenovirus-mediated Gene Transfer of MMP-2 into Cultured Porcine Trabecular Meshwork Cells

This study aimed to use adenoviral gene transfer to express matrix metalloproteinase (MMP)-2 in cultured porcine trabecular meshwork cells and to evaluate the duration of adenovirus-mediated MMP-2 expression and its enzymatic activity. MMP-2 cDNA was synthesized by ligating three segments of MMP-2 cDNA obtained by reverse transcription-polymerase chain reaction (RT-PCR) with mRNA extracted from mouse lungs. MMP-2 cDNA was inserted into replication-deficient adenoviral vectors. Western blotting revealed that MMP-2 was highly expressed by adenoviral gene transfer in cultured porcine trabecular meshwork cells. Zymography confirmed that the expressed MMP-2 possessed enzymatic activity and that MMP-2 activity increased dose-responsively with the viral titer. MMP-2 expression was detected two days after the additional virus preparation and continued for at least three weeks. Adenoviral vectors could efficiently deliver MMP-2 cDNA to cultured trabecular meshwork cells, with MMP-2 gene expression persisting for three weeks after infection. Our data have implications for future gene therapy in glaucoma.