title: In vitro characterization of the angiogenic phenotype and genotype of the endothelia derived from sporadic cerebral cavernous malformations.
abstract: BACKGROUND
  Cerebral cavernous malformation (CCM) is mainly a disorder of endothelial cells. Although the endothelial function of CCM genes has been characterized in familial CCMs, little attention has been paid to the pathological alterations of the endothelium in sporadic CCMs.
  
  
  OBJECTIVE
  We assumed that the endothelia derived from sporadic CCMs present genotypic and/or phenotypic alterations and exhibit unique responses to the pathogenic stimuli.
  
  
  METHODS
  Endothelial cells were prepared from fresh operative specimens of sporadic CCMs with a single lesion (CCM-ECs, n = 20). The expression of VEGF and its receptors and CCM1-3 genes were detected by real-time reverse transcriptase polymerase chain reaction (RT-PCR). Endothelial cell proliferation, migration, sprouting, and tube formation were compared between CCM-ECs and control endothelial cells after different angiogenic stimuli and after silencing CCM1.
  
  
  RESULTS
  RT-PCR revealed a highly activated VEGF system in CCM-ECs without significant alteration in CCM1-3 gene expression. Accordingly, CCM-ECs exhibited great growth potential under normal culture conditions and a significantly high proliferation activity in response to various angiogenic stimuli including hypoxia, fetal calf serum, and vascular endothelial growth factor treatment. A considerably higher mobility, spontaneous sprouting and extensive tube-branching were exclusively detected in CCM-ECs. In comparison with control endothelia, CCM-EC resisted apoptotic stimuli and showed distinct responses to activating angiogenesis after silencing CCM1.
  
  
  CONCLUSIONS
  Distinct genotypic and phenotypic features occur in CCM-EC independently from the deficiency in CCM1-3 gene expression. The distinct responses of CCM-EC to different pathogenic stimuli suggest that CCM-EC is a valuable in vitro model for further study of CCMs.
authors.affiliation: Department of Neurosurgery, University of Duisburg-Essen, Essen, Germany. yuan.zhu@uk-essen.de
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhu
authors.forename: Qun
authors.initials: Q
authors.lastname: Wu
authors.forename: Moritz
authors.initials: M
authors.lastname: Fass
authors.forename: Jin-Fang
authors.initials: JF
authors.lastname: Xu
authors.forename: Chao
authors.initials: C
authors.lastname: You
authors.forename: Oliver
authors.initials: O
authors.lastname: Müller
authors.forename: I Erol
authors.initials: IE
authors.lastname: Sandalcioglu
authors.forename: Jian-Min
authors.initials: JM
authors.lastname: Zhang
authors.forename: Ulrich
authors.initials: U
authors.lastname: Sure
mesh_terms: D000328:Adult; D017209:Apoptosis / Q000502:physiology; D015687:Cell Hypoxia; D002465:Cell Movement / Q000502:physiology; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D049109:Cell Proliferation; D002469:Cell Separation; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D042783:Endothelial Cells / Q000502:physiology; D004730:Endothelium, Vascular / Q000473:pathology*; D005260:Female; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D020868:Gene Silencing; D005838:Genotype; D020786:Hemangioma, Cavernous, Central Nervous System / Q000235:genetics* / Q000473:pathology*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007150:Immunohistochemistry; D002538:Intracranial Arteriovenous Malformations / Q000235:genetics* / Q000473:pathology*; D008297:Male; D009389:Neovascularization, Pathologic / Q000235:genetics*; D009924:Organ Culture Techniques; D010641:Phenotype; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D018076:DNA, Complementary; D012313:RNA