Corneal angiogenesis modulation by cysteine cathepsins: in vitro and in vivo studies

Corneal angiogenesis modulation by cysteine cathepsins: in vitro and in vivo studies

Author Coppini, Larissa Pereira Autor UNIFESP Google Scholar
Visniauskas, Bruna Autor UNIFESP Google Scholar
Costa, Elaine F. Google Scholar
Filho, Milton N. Autor UNIFESP Google Scholar
Rodrigues, Eduardo B. Autor UNIFESP Google Scholar
Chagas, Jair R. Autor UNIFESP Google Scholar
Farah, Michel E. Autor UNIFESP Google Scholar
Barros, Nilana M. T. Autor UNIFESP Google Scholar
Carmona, Adriana K. Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Univ Fed Maranhao
Abstract Corneal avascularization is essential for normal vision. Several antiangiogenic factors were identified in cornea such as endostatin and angiostatin. Cathepsin V, which is highly expressed in the cornea, can hydrolyze human plasminogen to release angiostatin fragments. Herein, we describe a detailed investigation of the expression profile of cathepsins B, L, S and V in the human cornea and the role of cysteine peptidases in modulating angiogenesis both in vitro and in vivo. We used various methodological tools for this purpose, including real-time PCR, SDS-PAGE, western blotting, catalytic activity assays, cellular assays and induction of corneal neovascularity in rabbit eyes. Human corneal enzymatic activity assays revealed the presence of cysteine proteases that were capable of processing endogenous corneal plasminogen to produce angiostatin-like fragments. Comparative real-time analysis of cathepsin B, L, S and V expression revealed that cathepsin V was the most highly expressed, followed by cathepsins L, B and S. However, cathepsin V depletion revealed that this enzyme is not the major cysteine protease responsible for plasminogen degradation under non-pathological conditions. Furthermore, western blotting analysis indicated that only cathepsins B and S were present in their enzymatically active forms. in vivo analysis of angiogenesis demonstrated that treatment with the cysteine peptidase inhibitor E64 caused a reduction in neovascularization. Taken together, our results show that human corneal cysteine proteases are critically involved in angiogenesis. (C) 2015 Elsevier B.V. All rights reserved.
Keywords Cysteine proteases
Plasminogen
Ocular angiogenesis
Cathepsins
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Grant number CNPq: 482400/2013-7
Date 2015-05-01
Published in Experimental Eye Research. London: Academic Press Ltd- Elsevier B.V., v. 134, p. 39-46, 2015.
ISSN 0014-4835 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 39-46
Origin http://dx.doi.org/10.1016/j.exer.2015.03.012
Access rights Closed access
Type Article
Web of Science ID WOS:000354142000005
URI http://repositorio.unifesp.br/handle/11600/39043

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