Actions of the Kunitz-type serine protease inhibitor Amblyomin-X on VEGF-A-induced angiogenesis

C C Drewes; R Y S Dias; C B Hebeda; S M Simons; S A Barreto; J M Ferreira Jr; A M Chudzinski-Tavassi; S H P Farsky

doi:10.1016/j.toxicon.2012.04.349

Actions of the Kunitz-type serine protease inhibitor Amblyomin-X on VEGF-A-induced angiogenesis

Toxicon. 2012 Sep 1;60(3):333-40. doi: 10.1016/j.toxicon.2012.04.349. Epub 2012 May 7.

Authors

C C Drewes¹, R Y S Dias, C B Hebeda, S M Simons, S A Barreto, J M Ferreira Jr, A M Chudzinski-Tavassi, S H P Farsky

Affiliation

¹ Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of Sao Paulo, Sao Paulo, SP, Brazil.

PMID: 22575283
DOI: 10.1016/j.toxicon.2012.04.349

Abstract

Amblyomin-X is a Kunitz-type serine protease inhibitor (Kunitz-type SPI) designed from the cDNA library of the Amblyomma cajennense tick, which displays in vivo anti-tumor activities. Here, the mechanisms of actions of Amblyomin-X in vascular endothelial growth factor A (VEGF-A)-induced angiogenesis were characterized. Topical application of Amblyomin-X (10 or 100 ng/10 μl; each 48 h) inhibited VEGF-A-induced (10 ng/10 μl; each 48 h) angiogenesis in the dorsal subcutaneous tissue in male Swiss mice. Moreover, similar effect was observed in the VEGF-A-induced angiogenesis in the chicken chorioallantoic membrane (CAM). Additional in vitro assays in t-End cells showed that Amblyomin-X treatment delayed the cell cycle, by maintaining them in G0/G1 phase, and inhibited cell proliferation and adhesion, tube formation and membrane expression of the adhesion molecule platelet-endothelial cell adhesion molecule-1 (PECAM-1), regardless of mRNA synthesis. Together, results herein reveal the role of Kunitz-type SPI on in vivo VEGF-A-induced angiogenesis, by exerting modulatory actions on endothelial cell proliferation and adhesion, especially on membrane expression of PECAM-1. These data provide further mechanisms of actions of Kunitz-type SPI, corroborating their relevance as scientific tools in the design of therapeutic molecules.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Angiogenesis Inhibitors / genetics
Angiogenesis Inhibitors / metabolism
Angiogenesis Inhibitors / pharmacology*
Animals
Arthropod Proteins / genetics
Arthropod Proteins / metabolism
Arthropod Proteins / pharmacology
Cell Adhesion / drug effects
Cell Line, Transformed
Cell Proliferation / drug effects
Chick Embryo
Chorioallantoic Membrane / drug effects
Chorioallantoic Membrane / metabolism
Endothelial Cells / drug effects*
Endothelial Cells / metabolism
Factor Xa Inhibitors
Ixodidae / metabolism
Male
Mice
Neovascularization, Pathologic / drug therapy*
Neovascularization, Physiologic / drug effects*
Platelet Endothelial Cell Adhesion Molecule-1 / genetics
Platelet Endothelial Cell Adhesion Molecule-1 / metabolism
Recombinant Proteins / metabolism
Recombinant Proteins / pharmacology
Resting Phase, Cell Cycle / drug effects
Salivary Proteins and Peptides / genetics
Salivary Proteins and Peptides / metabolism
Salivary Proteins and Peptides / pharmacology*
Serine Proteinase Inhibitors / pharmacology*
Subcutaneous Tissue / drug effects
Subcutaneous Tissue / metabolism
Vascular Endothelial Growth Factor A / metabolism*

Substances

Amblyomin-X protein, Amblyomma cajennense
Angiogenesis Inhibitors
Arthropod Proteins
Factor Xa Inhibitors
Platelet Endothelial Cell Adhesion Molecule-1
Recombinant Proteins
Salivary Proteins and Peptides
Serine Proteinase Inhibitors
Vascular Endothelial Growth Factor A