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Effect of Crotoxin on several steps in angiogenese evaluated in two dimensional and three dimensional matrices: in vitro studies

Grant number: 12/51241-5
Support Opportunities:Regular Research Grants
Start date: February 01, 2013
End date: January 31, 2015
Field of knowledge:Biological Sciences - Pharmacology - General Pharmacology
Principal Investigator:Sandra Coccuzzo Sampaio Vessoni
Grantee:Sandra Coccuzzo Sampaio Vessoni
Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

Several studies, in vivo and in vitro have demonstrated antitumor activity of Crotoxin (CTX), the major toxin of Crotalus durissus terrificus venom. Angiogenesis plays a key role in tumorigenesis and metastatic process. The formation of new blood vessels is the principal process of angiogenesis and involves adhesion, proliferation and migration of endothelial cells to reach remote targets, assembly of endothelial cells into new capillary tubes. Despite evidence of antitumor action of crotoxin, was not demonstrated yet the action of this toxin on basic parameters for neovascularization, essential for the growth and survival of the tumor. Thus, studies demonstrating the action of crotoxin on basic parameters for neovascularization may contribute to the characterization of the antitumor action of CTX and contribute to the elucidation of novel mechanisms of action. This study aims to evaluate the effects of CTX on the invasion and migration in vitro endothelial cell lineage thymic (tEnd.1) systems using two-dimensional (2-D type I collagen gel containing fibronectin) and three-dimensional (3-D I collagen gel containing fibronectin). The following parameters are determined: 1) the action of CTX on cell invasion t.END.1 extracellular matrix by means of adhesion behavior of endothelial cells to their natural ligands, such as collagen 1, fibronectin, and fibrinogen, using competition assay and migration evaluated by the Wound healing, 2) The migratory behavior (speed and type of migration) cells t.End.1 substrate in 2-D and 3-D collagen type I containing fibronectin, assessed by real-time technique of time-lapse, 3) on the action of CTX induced vessel formation in matrigel in the 3D model, 4) the expression of Rho GTPases (RhoA, Cdc42 and Rac 1) and integrin α2β1 t.END.1 cells evaluated by immunocytochemistry assays; 5) release of metalloproteinases (MMP-2 and MMP-9) and VEGF, after treatment with crotoxin, by enzyme immunoassay (EIA). (AU)

Articles published in Agência FAPESP Newsletter about the research grant:
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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
BUFALO, MICHELLE C.; ALMEIDA, MAIRA E.; FRANCA, ISABELLA ARAUJO; ZAMBELLI, VANESSA O.; SANT'ANNA, MORENA BRAZIL MARTINS; KIMURA, LOUISE F.; GIARDINI, ALINE CAROLINA; CURY, YARA; SAMPAIO, SANDRA COCCUZZO. Advanced glycation endproducts produced by in vitro glycation of type I collagen modulate the functional and secretory behavior of dorsal root ganglion cells cultivated in two-dimensional system. Experimental Cell Research, v. 382, n. 2-3, . (16/12128-0, 15/50040-4, 12/51241-5, 16/10886-4)
KATO, ELLEN EMI; PIMENTA, LUCIANA ARAUJO; SOARES DE ALMEIDA, MAIRA ESTANISLAU; ZAMBELLI, VANESSA OLZON; DOS SANTOS, MARINILCE FAGUNDES; SAMPAIO, SANDRA COCCUZZO. rotoxin Inhibits Endothelial Cell Functions in Two- and Three-dimensional Tumor Microenvironmen. FRONTIERS IN PHARMACOLOGY, v. 12, . (13/07467-1, 12/51241-5)
PIMENTA, LUCIANA DE ARAUJO; DE ALMEIDA, MAIRA ESTANISLAU S.; BRETONES, MARISA LANGEANI; CIRILLO, MARIA CRISTINA; CURI, RUI; SAMPAIO, SANDRA COCCUZZO. Crotoxin promotes macrophage reprogramming towards an antiangiogenic phenotype. SCIENTIFIC REPORTS, v. 9, . (13/17190-7, 08/57898-0, 12/51241-5)