TGF-beta2-induced matrix modification and cell transdifferentiation in the human lens capsular bag.

Abstract

PURPOSE:

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.

• Authors: I Michael Wormstone, Shigeo Tamiya, Ian Anderson, George Duncan
• Journal: Investigative ophthalmology & visual science (Invest Ophthalmol Vis Sci) Vol. 43 Issue 7 Pg. 2301-8 (Jul 2002) ISSN: 0146-0404 [Print] United States
• PMID: 12091431 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Actins
• Antibodies, Monoclonal
• Fibronectins
• TGFB2 protein, human
• Transforming Growth Factor beta
• Transforming Growth Factor beta2
• Matrix Metalloproteinase 2
• Matrix Metalloproteinase 9

Topics

• Actins (metabolism)
• Antibodies, Monoclonal (therapeutic use)
• Cataract (etiology, metabolism, pathology, prevention & control)
• Cataract Extraction
• Cell Differentiation (drug effects)
• Enzyme-Linked Immunosorbent Assay
• Extracellular Matrix (metabolism)
• Fibronectins (metabolism)
• Fluorescent Antibody Technique, Indirect
• Humans
• Lens Capsule, Crystalline (drug effects, metabolism, pathology)
• Lens Implantation, Intraocular
• Matrix Metalloproteinase 2 (metabolism)
• Matrix Metalloproteinase 9 (metabolism)
• Organ Culture Techniques
• Transforming Growth Factor beta (immunology, metabolism, pharmacology)
• Transforming Growth Factor beta2