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Abstract #18335 Published in IGR 3-3
Stromelysin gene transfer into cultured human trabecular cells and rat trabecular meshwork in vivo
Kee C; Sohn S; Hwang JMInvestigative Ophthalmology and Visual Science 2001; 42: 2856-2860
PURPOSE: To determine whether stromelysin gene can be introduced into and expressed in cultured human trabecular cells as well as in the rat eye in vivo by means of a recombinant replication-deficient adenovirus. METHODS: Stromelysin cDNA was obtained by reverse transcription-polymerase chain reaction with mRNA extracted from cultured human trabecular cells after induction with interleukin 1α. Adenovirus vector containing stromelysin cDNA was constructed by cotransfection of pJM17 and PΔA.CMV-str into the 293 cells. The expression of stromelysin in the cultured human trabecular cells was assayed by western blot and zymography. The expression 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 El region. Western blot and zymogram revealed that the stromelysin could be expressed and that it possessed enzymatic activity in cultured human trabecular cells. In situ hybridization and immunostaining of the stromelysin showed that the complete form of stromelysin was expressed in the trabecular meshwork, iris, and uveoscleral outflow pathway of the rat eye. CONCLUSIONS: Stromelysin, a functional gene, can be transferred in vivo into rat eyes and in vitro into cultured human trabecular cells using a replication-deficient adenovirus vector. This shows the possibility of gene therapy in glaucoma.
Dr C. Kee, Department of Ophthalmology, Samsung Medical Center, Sung Kyun Kwan University School of Medicine, 50 Hwon-dong Kangnam-ku, Seoul 135-710, Korea. cwkee@ophthalmology.org
Classification:
11.9 Gene therapy (Part of: 11 Medical treatment)