title: Anti-angiogenic activities of snake venom CRISP isolated from Echis carinatus sochureki.
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.
  
  
  CONCLUSIONS
  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: Temple University, College of Engineering, Department of Bioengineering, Philadelphia, PA, USA.
authors: Shimon
authors: S
authors: Lecht
authors: Temple University, College of Engineering, Department of Bioengineering, Philadelphia, PA, USA.
authors: Rachel A
authors: RA
authors: Chiaverelli
authors: Temple University, College of Engineering, Department of Bioengineering, Philadelphia, PA, USA.
authors: Jonathan
authors: J
authors: Gerstenhaber
authors: Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Cientificas, 46010 Valencia, Spain.
authors: Juan J
authors: JJ
authors: Calvete
authors: The Hebrew University of Jerusalem, School of Pharmacy, Institute for Drug Research, Jerusalem, Israel.
authors: Philip
authors: P
authors: Lazarovici
authors: The Alistar Reid Venom Research Unit, Liverpool School of Tropical Medicine, Liverpool L3 5QA, United Kingdom.
authors: Nicholas R
authors: NR
authors: Casewell
authors: The Alistar Reid Venom Research Unit, Liverpool School of Tropical Medicine, Liverpool L3 5QA, United Kingdom.
authors: Robert
authors: R
authors: Harrison
authors: Temple University, College of Engineering, Department of Bioengineering, Philadelphia, PA, USA.
authors: Peter I
authors: PI
authors: Lelkes
authors: Temple University, College of Engineering, Department of Bioengineering, Philadelphia, PA, USA. Electronic address: cmarcink@temple.edu.
authors: Cezary
authors: C
authors: Marcinkiewicz
mesh_terms: D000595:Amino Acid Sequence; D020533:Angiogenesis Inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D042501:Angiogenic Proteins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D049033:Chorioallantoic Membrane / Q000098:blood supply*; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D005910:Glioma / Q000098:blood supply* / Q000473:pathology; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009389:Neovascularization, Pathologic; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D011487:Protein Conformation; D011784:Quail; D015398:Signal Transduction / Q000187:drug effects; D014757:Viper Venoms / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D020533:Angiogenesis Inhibitors; D042501:Angiogenic Proteins; C000600681:CRISP protein, Echis carinatus sochureki; D014757:Viper Venoms
keywords: Angiogenesis; CRISP; Cell proliferation; Endothelial cells; Migration