TY - JOUR
T1 - Anti-connective tissue growth factor antibody treatment reduces Extracellular matrix production in Trabecular meshwork and Lamina Cribrosa cells
AU - Wallace, Deborah M.
AU - Clark, Abbot F.
AU - Lipson, Kenneth E.
AU - Andrews, Darrell
AU - Crean, John K.
AU - O'Brien, Colm J.
PY - 2013/11/7
Y1 - 2013/11/7
N2 - Purpose. We have previously demonstrated elevated levels of connective tissue growth factor (CTGF/CCN2) in the aqueous humor (AqH) of pseudoexfoliation glaucoma (PXFG) patients when compared with cataract controls. Furthermore, there is a significant trabecular meshwork (TM) and lamina cribrosa (LC) fibrotic phenotype associated with glaucoma, possibly driven by CTGF. The purpose of this study was to investigate the potential of anti-CTGF immunotherapy in glaucoma. Methods. Primary TM and LC cells were cultured from human donors with (GTM/GLC) and without (NTM/NLC) primary open angle glaucoma (POAG). Aqueous humor samples from PXFG, POAG, and control cataract patients were applied to N/GTM and N/GLC cells in the presence or absence of a therapeutic, humanized monoclonal anti-CTGF antibody FG-3019 (10 lg/mL). Hydrogen peroxide (H2O2) was also used as a stimulus. Expression of fibrotic genes (fibronectin-1, fibrillin-1, CTGF, collagen type I a1, and a-smooth muscle actin) was assessed by q-PCR. Protein expression of collagen 1A1 and a-smooth muscle actin was examined in N/G TM cells by SDS-PAGE. The modulatory effect of FG-3019 (10 lg/mL) and IgG (10 lg/mL) were also assessed. Results. Treatment of cells with AqH from PXFG and POAG patients and H2O2 induced a significant (P < 0.05) increase in expression of profibrotic genes, which was significantly reduced by pretreatment with FG-3019 (P < 0.05). FG-3019 also reduced expression of a-smooth muscle actin and collagen 1A1 protein expression in N/GTM cells. Conclusions. FG-3019 is effective in blocking extracellular matrix production in TM and LC cells, thus supporting a role for the use of anti-CTGF immunotherapy in the treatment of glaucoma. 2013 The Association for Research in Vision and Ophthalmology, Inc.
AB - Purpose. We have previously demonstrated elevated levels of connective tissue growth factor (CTGF/CCN2) in the aqueous humor (AqH) of pseudoexfoliation glaucoma (PXFG) patients when compared with cataract controls. Furthermore, there is a significant trabecular meshwork (TM) and lamina cribrosa (LC) fibrotic phenotype associated with glaucoma, possibly driven by CTGF. The purpose of this study was to investigate the potential of anti-CTGF immunotherapy in glaucoma. Methods. Primary TM and LC cells were cultured from human donors with (GTM/GLC) and without (NTM/NLC) primary open angle glaucoma (POAG). Aqueous humor samples from PXFG, POAG, and control cataract patients were applied to N/GTM and N/GLC cells in the presence or absence of a therapeutic, humanized monoclonal anti-CTGF antibody FG-3019 (10 lg/mL). Hydrogen peroxide (H2O2) was also used as a stimulus. Expression of fibrotic genes (fibronectin-1, fibrillin-1, CTGF, collagen type I a1, and a-smooth muscle actin) was assessed by q-PCR. Protein expression of collagen 1A1 and a-smooth muscle actin was examined in N/G TM cells by SDS-PAGE. The modulatory effect of FG-3019 (10 lg/mL) and IgG (10 lg/mL) were also assessed. Results. Treatment of cells with AqH from PXFG and POAG patients and H2O2 induced a significant (P < 0.05) increase in expression of profibrotic genes, which was significantly reduced by pretreatment with FG-3019 (P < 0.05). FG-3019 also reduced expression of a-smooth muscle actin and collagen 1A1 protein expression in N/GTM cells. Conclusions. FG-3019 is effective in blocking extracellular matrix production in TM and LC cells, thus supporting a role for the use of anti-CTGF immunotherapy in the treatment of glaucoma. 2013 The Association for Research in Vision and Ophthalmology, Inc.
KW - Extracellular matrix
KW - Glaucoma pharmacology
KW - Lamina cribrosa
KW - Trabecular meshwork
UR - http://www.scopus.com/inward/record.url?scp=84888873912&partnerID=8YFLogxK
U2 - 10.1167/iovs.13-12494
DO - 10.1167/iovs.13-12494
M3 - Article
C2 - 24204045
AN - SCOPUS:84888873912
SN - 0146-0404
VL - 54
SP - 7836
EP - 7848
JO - Investigative Ophthalmology and Visual Science
JF - Investigative Ophthalmology and Visual Science
IS - 13
ER -