Abstract
Purpose: :
We have previously demonstrated elevated levels of connective tissue growth factor (CTGF) in the aqueous humour of Pseudoexfoliation glaucoma (PXFG) and Primary open angle glaucoma (POAG) patients compared to controls. CTGF was also shown to upregulate fibrillin-1 (a component of PXF-material) expression in cultured human trabecular meshwork (TM) cells. The purpose of this study was to investigate the therapeutic potential of anti-CTGF immunotherapy in PXFG and POAG in combating fibrosis using oxidative stress and aqueous humour samples as stimuli in primary TM and primary lamina cribrosa (LC) cells.
Methods: :
Aqueous humour samples from PXFG, POAG and control patients (n=3) were applied to porcine TM and human LC cells in the presence or absence of the monoclonal anti-CTGF antibody FG-3019 (10μM) (FibroGen Inc, South San Francisco, CA). Hydrogen peroxide (200μM H2O2, a subtoxic dose as determined by crystal violet assay) was used as a glaucoma-like (oxidative stress) stimulus +/- FG-3019. CTGF, fibrillin-1 and fibronectin-1 expression levels were assessed by quantitative PCR using gene specific exon-exon spanning primers. IgG antibody (10μM) was included as a negative control in both sets of experiments.
Results: :
Results showed that incubation of LC and TM cells with aqueous humour from both PXFG and POAG patients induced a significant (P<0.05) increase in expression of CTGF, fibrillin 1 and fibronectin 1. This fibrotic response was significantly reduced by the inclusion of FG-3019 (P<0.05). Similar results were obtained for oxidative stress treatment of LC and TM cells (P<0.05).
Conclusions: :
The anti-CTGF antibody FG-3019 is effective in blocking CTGF, fibrillin 1 and fibronectin 1 production. These observed anti-fibrotic effects support a pathologically significant role for the use of anti-CTGF immunotherapy as a possible approach for treatment of PXFG and POAG.
Keywords: extracellular matrix • trabecular meshwork • lamina cribrosa