TGF-beta2-induced matrix modification and cell transdifferentiation in the human lens capsular bag.
OBJECTIVE
To study the role of TGF-beta2 in posterior capsule opacification (PCO) and to determine whether CAT-152 (lerdelimumab), a fully human monoclonal antibody that neutralizes the effect of TGF-beta2, can also provide therapeutic benefit for PCO.


METHODS
In vitro capsular bags were prepared from human donor eyes and maintained in a 5% CO(2) atmosphere at 35 degrees C. To investigate expression of active TGF-beta2, capsular bags were incubated in serum-free EMEM for 2, 28, or more than 100 days and analyzed by ELISA (n > or = 4 at each time point). To study underlying mechanisms, match-pair experiments were also performed, so that the medium was supplemented with 0, 1 or 10 ng/mL TGF-beta2 with or without 10 microg/mL CAT-152 (n = 4 in all cases). On-going observations were by phase-contrast microscopy. In addition, donor material from patients who had undergone cataract surgery was analyzed. Cellular architecture was examined by fluorescence cytochemistry. Expression of matrix metalloproteinase (MMP)-2 and -9 was assessed by gelatin zymography.


RESULTS
Analysis of capsular bags from donor eyes that had received an intraocular lens (IOL) revealed the presence of endogenous active TGF-beta2, matrix wrinkling, and expression of transdifferentiation markers alphaSMA and fibronectin. When cultured in vitro, donor bags also showed sustained release of MMP-2 and -9. Culture of capsular bags prepared in vitro from whole lenses showed that TGF-beta2 (1-10 ng/mL) stimulated transdifferentiation and contraction of the capsular bag, resulting in light scatter. TGF-beta2 also induced sustained release of MMP-2 and -9. Active TGF-beta2 was detected in these cultures. The human monoclonal anti-TGF-beta2 antibody CAT-152 (10 microg/mL) effectively inhibited all TGF-beta2-induced effects.


CONCLUSIONS
Addition of TGF-beta2 accelerates transdifferentiation and contraction of the capsular bag, resulting in light scatter. CAT-152 inhibited all the effects of TGF-beta2 that were examined and therefore has the potential to suppress development of PCO and provide potential therapeutic benefit to cataract patients.
Wormstone
I Michael
IM
School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK. i.m.wormstone@uea.ac.uk
Tamiya
Shigeo
S
Anderson
Ian
I
Duncan
George
G
D000199:Actins / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000627:therapeutic use; D002386:Cataract / Q000209:etiology* / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control; D002387:Cataract Extraction; D002454:Cell Differentiation / Q000187:drug effects*; D004797:Enzyme-Linked Immunosorbent Assay; D005109:Extracellular Matrix / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007903:Lens Capsule, Crystalline / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D019654:Lens Implantation, Intraocular; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D009924:Organ Culture Techniques; D016212:Transforming Growth Factor beta / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D053781:Transforming Growth Factor beta2
D000199:Actins; D000911:Antibodies, Monoclonal; D005353:Fibronectins; C509041:TGFB2 protein, human; D016212:Transforming Growth Factor beta; D053781:Transforming Growth Factor beta2; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9