gms | German Medical Science

61st Annual Meeting of the German Society of Neurosurgery (DGNC) as part of the Neurowoche 2010
Joint Meeting with the Brazilian Society of Neurosurgery on the 20 September 2010

German Society of Neurosurgery (DGNC)

21 - 25 September 2010, Mannheim

Loss of CCM2, but not CCM3, impairs the integrity of endothelial tube structure – An association with the incidence of intracerebal haemorrhage of inherited cerebral cavernous malformations (CCMs)?

Meeting Abstract

  • Yuan Zhu - Klinik für Neurochirurgie, Universitätsklinikum Essen, Deutschland
  • Jin-Fang Xu - Klinik für Neurochirurgie, Universitätsklinikum Essen, Deutschland
  • Dorothea Miller - Klinik für Neurochirurgie, Universitätsklinikum Essen, Deutschland
  • I. Erol Sandalcioglu - Klinik für Neurochirurgie, Universitätsklinikum Essen, Deutschland
  • Ulrich Sure - Klinik für Neurochirurgie, Universitätsklinikum Essen, Deutschland

Deutsche Gesellschaft für Neurochirurgie. 61. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC) im Rahmen der Neurowoche 2010. Mannheim, 21.-25.09.2010. Düsseldorf: German Medical Science GMS Publishing House; 2010. DocP1839

doi: 10.3205/10dgnc310, urn:nbn:de:0183-10dgnc3106

Published: September 16, 2010

© 2010 Zhu et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.



Objective: Mutations in CCM genes may cause autosomal dominant cerebral cavernous malformations (CCMs). Indeed, a different intracranial hemorrhage rate has been observed in CCMs carrying individual CCM gene mutations. We proposed a differential regulation of angiogenesis by these disease genes. The present study thus investigated the function of CCM2 and CCM3 in tube formation, a key step involved in angiogenesis.

Methods: CCM2 and CCM3 were silenced by the specific siRNAs in endothelial cells (HUVEC). The time course of the gene silencing was established by detecting the efficiency of knockdown using real-time PCR. For tube formation assay, the transfected cells were pooled in matrigel-coated 96-well plate followed by 6 h, 24 h and 48 h of incubation. The tube formation was analyzed under a microscope.

Results: Real-time PCR showed that CCM2 expression declined by more that 80% at 48 h and 72 h, and returned to the control level at 96 h after the transfection, whereas CCM3 gene levels were consistently reduced around 80% from 48 h to 96 h after siRNA. Under these efficient knockdown conditions, we observed that the tube was similarly formed in the CCM2- and CCM3-silenced cells and in control at 6h after the incubation. However, the maintenance of tube integrity was largely impaired by CCM2, but not CCM3, silencing at 24 h and 48 h after pooling cells on matrigel. Quantification of the number of tubes revealed that CCM2 silencing resulted in a 50% of reduction of tubes at 24 h and a complete loss of the tube structure at 48h after the incubation.

Conclusions: Our results demonstrated a distinct role of CCM2 and CCM3 in maintenance of tube stability. The severe impairment of the integrity of tube structure after endothelial CCM2 silencing may be associated with the clinical observation that CCM2-mutated CCMs present higher incidence of haemorrhage in comparison to CCM3-mutated cases.