Development of gene theropy for glaucoma by neaws of modulation of proteoglycan praduction in aqueous outflow rocite

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We have investigated our hypothesis on mechanism of primary angly open glaucoma, that an interactive relationship between TGF-beta2 in aqueous humor and proteoglycan (PG) expression in trabecular meshwork (TM) cells, may play crucial roles in regulation of aqueous outflow resistance.Gene expression of the PG core protein was screened in human and bovine cultured TM cells by reverse transcription -initiated polymerase chain reaction (RT-PCR) and the RT-PCR experiments showed that cDNA fragments of the three PGs (decorin, lumican and fibromodulin). Subsequently, the TM cells were cultured with TGF-beta2, and the changes of gene expressions of the PGs were investigated. As a result, the expression of three PGs (decorin, lumican and fibromodulin) were up-regulated by TGF-beta2 stimulation.In addition, we assessed TGF-beta2 in aqueous humor which were collected from variable glaucomatous patients and non-glaucomatous (cataractous) patients with a specific-capture ELISA.Our study revealed that increased levels of TGF-beta2 in aqueous humor was found only in eyes with POAG,but not with EXS and PACG.These findings suggested that increased levels of TGF-beta2 may be different from other types of glaucoma.Furthrmore, we attempted to create a POAG model by means of TGF-beta2 gene transfer and resultant increase of proteolglycan production in TM cells, and in autocrine and/or paracrine manners of production. We have succeeded to efficiently introduce genes to primate and human TM cells by the HVJ liposome method. This results showed the potential of this method for a somatic gene therapy of the trabecular meshwork to treat glaucoma.These result suggested the potential role of increased levels of TGF-beta2 in aqueous humor and resultant increase of proteolglycan production in the pathogenesis of primary open angle glaucoma, and the gene transfer methods may be useful to treat glaucoma.