Shimon Lecht1, Rachel A Chiaverelli1, Jonathan Gerstenhaber1, Juan J Calvete2, Philip Lazarovici3, Nicholas R Casewell4, Robert Harrison4, Peter I Lelkes1, Cezary Marcinkiewicz5. 1. Temple University, College of Engineering, Department of Bioengineering, Philadelphia, PA, USA. 2. Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Cientificas, 46010 Valencia, Spain. 3. The Hebrew University of Jerusalem, School of Pharmacy, Institute for Drug Research, Jerusalem, Israel. 4. The Alistar Reid Venom Research Unit, Liverpool School of Tropical Medicine, Liverpool L3 5QA, United Kingdom. 5. Temple University, College of Engineering, Department of Bioengineering, Philadelphia, PA, USA. Electronic address: cmarcink@temple.edu.
Abstract
BACKGROUND: Cysteine-rich secretory protein (CRISP) is present in majority of vertebrate including human. The physiological role of this protein is not characterized. We report that a CRISP isolated from Echis carinatus sochureki venom (ES-CRISP) inhibits angiogenesis. METHODS: The anti-angiogenic activity of purified ES-CRISP from snake venom was investigated in vitro using endothelial cells assays such as proliferation, migration and tube formation in Matrigel, as well as in vivo in quail embryonic CAM system. The modulatory effect of ES-CRISP on the expression of major angiogenesis factors and activation of angiogenesis pathways was tested by qRT-PCR and Western blot. RESULTS: The amino acid sequence of ES-CRISP was found highly similar to other members of this snake venom protein family, and shares over 50% identity with human CRISP-3. ES-CRISP supported adhesion to endothelial cells, although it was also internalized into the cytoplasm in a granule-like manner. It blocked EC proliferation, migration and tube formation in Matrigel. In the embryonic quail CAM system, ES-CRISP abolished neovascularization process induced by exogenous growth factors (bFGF, vpVEGF) and by developing gliomas. CRISP modulates the expression of several factors at the mRNA level, which were characterized as regulators of angiogenesis and blocked activation of MAPK Erk1/2 induced by VEGF. CONCLUSIONS: ES-CRISP was characterized as a negative regulator of the angiogenesis, by direct interaction with endothelial cells. GENERAL SIGNIFICANCE: The presented work may lead to the development of novel angiostatic therapy, as well as contribute to the identification of the physiological relevance of this functionally uncharacterized protein.
BACKGROUND: Cysteine-rich secretory protein (CRISP) is present in majority of vertebrate including human. The physiological role of this protein is not characterized. We report that a CRISP isolated from Echis carinatus sochureki venom (ES-CRISP) inhibits angiogenesis. METHODS: The anti-angiogenic activity of purified ES-CRISP from snake venom was investigated in vitro using endothelial cells assays such as proliferation, migration and tube formation in Matrigel, as well as in vivo in quail embryonic CAM system. The modulatory effect of ES-CRISP on the expression of major angiogenesis factors and activation of angiogenesis pathways was tested by qRT-PCR and Western blot. RESULTS: The amino acid sequence of ES-CRISP was found highly similar to other members of this snake venom protein family, and shares over 50% identity with human CRISP-3. ES-CRISP supported adhesion to endothelial cells, although it was also internalized into the cytoplasm in a granule-like manner. It blocked EC proliferation, migration and tube formation in Matrigel. In the embryonic quail CAM system, ES-CRISP abolished neovascularization process induced by exogenous growth factors (bFGF, vpVEGF) and by developing gliomas. CRISP modulates the expression of several factors at the mRNA level, which were characterized as regulators of angiogenesis and blocked activation of MAPK Erk1/2 induced by VEGF. CONCLUSIONS: ES-CRISP was characterized as a negative regulator of the angiogenesis, by direct interaction with endothelial cells. GENERAL SIGNIFICANCE: The presented work may lead to the development of novel angiostatic therapy, as well as contribute to the identification of the physiological relevance of this functionally uncharacterized protein.
Authors: Montamas Suntravat; Walter E Cromer; Jessenia Marquez; Jacob A Galan; David C Zawieja; Peter Davies; Emelyn Salazar; Elda E Sánchez Journal: Toxicon Date: 2019-04-20 Impact factor: 3.033
Authors: Geoffrey Istas; Ken Declerck; Maria Pudenz; Katarzyna Szarc Vel Szic; Veronica Lendinez-Tortajada; Montserrat Leon-Latre; Karen Heyninck; Guy Haegeman; Jose A Casasnovas; Maria Tellez-Plaza; Clarissa Gerhauser; Christian Heiss; Ana Rodriguez-Mateos; Wim Vanden Berghe Journal: Sci Rep Date: 2017-07-11 Impact factor: 4.379
Authors: Marcelo R V Diniz; Ana L B Paiva; Clara Guerra-Duarte; Milton Y Nishiyama; Mauricio A Mudadu; Ursula de Oliveira; Márcia H Borges; John R Yates; Inácio de L Junqueira-de-Azevedo Journal: PLoS One Date: 2018-08-01 Impact factor: 3.240
Authors: Maja Lang Balija; Adrijana Leonardi; Marija Brgles; Dora Sviben; Tihana Kurtović; Beata Halassy; Igor Križaj Journal: Toxins (Basel) Date: 2020-03-16 Impact factor: 4.546
Authors: Montamas Suntravat; Oscar Sanchez; Armando Reyes; Abcde Cirilo; Jack S Ocheltree; Jacob A Galan; Emelyn Salazar; Peter Davies; Elda E Sanchez Journal: Toxins (Basel) Date: 2021-08-31 Impact factor: 4.546