OBJECTIVE: Germline mutations in 3 genes have been found in familial cases of cerebral cavernous malformations (CCMs). We previously discovered somatic and germline truncating mutations in the KRIT1 gene, supporting the "2-hit" mechanism of CCM lesion formation in a single lesion. The purpose of this study was to screen for somatic, nonheritable mutations in 3 more lesions from different patients and identify the cell type(s) in which somatic mutations occur. METHODS: Somatic mutations were sought in DNA from 3 surgically excised, fresh-frozen CCM lesions by cloning and screening polymerase chain reaction products generated from KRIT1 or PDCD10 coding regions. Laser capture microdissection was used on isolated endothelial and nonendothelial cells to determine whether somatic mutations were found in endothelial cells. RESULTS: CCM lesions harbor somatic and germline KRIT1 mutations on different chromosomes and are therefore biallelic. Both mutations are predicted to truncate the protein. The KRIT1 somatic mutations (novel c.1800delG mutation and previously identified 34 nucleotide deletion) in CCMs from 2 different patients were found only in the vascular endothelial cells lining caverns. No obvious somatic mutations were identified in the 2 other lesions; however, the results were inconclusive, possibly owing to the technical limitations or the fact that these specimens had a small proportion of vascular endothelial cells lining pristine caverns. CONCLUSION: The "2-hit" mechanism occurs in vascular endothelial cells lining CCM caverns from 2 patients with somatic and Hispanic-American KRIT1 germline mutations. Methods for somatic mutation detection should focus on vascular endothelial cells lining pristine caverns.
OBJECTIVE: Germline mutations in 3 genes have been found in familial cases of cerebral cavernous malformations (CCMs). We previously discovered somatic and germline truncating mutations in the KRIT1 gene, supporting the "2-hit" mechanism of CCM lesion formation in a single lesion. The purpose of this study was to screen for somatic, nonheritable mutations in 3 more lesions from different patients and identify the cell type(s) in which somatic mutations occur. METHODS: Somatic mutations were sought in DNA from 3 surgically excised, fresh-frozen CCM lesions by cloning and screening polymerase chain reaction products generated from KRIT1 or PDCD10 coding regions. Laser capture microdissection was used on isolated endothelial and nonendothelial cells to determine whether somatic mutations were found in endothelial cells. RESULTS: CCM lesions harbor somatic and germline KRIT1 mutations on different chromosomes and are therefore biallelic. Both mutations are predicted to truncate the protein. The KRIT1 somatic mutations (novel c.1800delG mutation and previously identified 34 nucleotide deletion) in CCMs from 2 different patients were found only in the vascular endothelial cells lining caverns. No obvious somatic mutations were identified in the 2 other lesions; however, the results were inconclusive, possibly owing to the technical limitations or the fact that these specimens had a small proportion of vascular endothelial cells lining pristine caverns. CONCLUSION: The "2-hit" mechanism occurs in vascular endothelial cells lining CCM caverns from 2 patients with somatic and Hispanic-American KRIT1 germline mutations. Methods for somatic mutation detection should focus on vascular endothelial cells lining pristine caverns.
Authors: T Watnick; N He; K Wang; Y Liang; P Parfrey; D Hefferton; P St George-Hyslop; G Germino; Y Pei Journal: Nat Genet Date: 2000-06 Impact factor: 38.330
Authors: Jon S Zawistowski; Ilya G Serebriiskii; Maximilian F Lee; Erica A Golemis; Douglas A Marchuk Journal: Hum Mol Genet Date: 2002-02-15 Impact factor: 6.150
Authors: C Denier; S Goutagny; P Labauge; V Krivosic; M Arnoult; A Cousin; A L Benabid; J Comoy; P Frerebeau; B Gilbert; J P Houtteville; M Jan; F Lapierre; H Loiseau; P Menei; P Mercier; J J Moreau; A Nivelon-Chevallier; F Parker; A M Redondo; J M Scarabin; M Tremoulet; M Zerah; J Maciazek; E Tournier-Lasserve Journal: Am J Hum Genet Date: 2004-01-22 Impact factor: 11.025
Authors: Christina L Liquori; Michel J Berg; Adrian M Siegel; Elizabeth Huang; Jon S Zawistowski; T'Prien Stoffer; Dominique Verlaan; Fiyinfolu Balogun; Lori Hughes; Tracey P Leedom; Nicholas W Plummer; Milena Cannella; Vittorio Maglione; Ferdinando Squitieri; Eric W Johnson; Guy A Rouleau; Louis Ptacek; Douglas A Marchuk Journal: Am J Hum Genet Date: 2003-11-17 Impact factor: 11.025
Authors: Alexander R Edelmann; Sarah Schwartz-Baxter; Christopher F Dibble; Warren C Byrd; Jim Carlson; Ivandario Saldarriaga; Sompop Bencharit Journal: Expert Rev Proteomics Date: 2014-03-31 Impact factor: 3.940
Authors: Aubrey C Chan; Stavros G Drakos; Oscar E Ruiz; Alexandra C H Smith; Christopher C Gibson; Jing Ling; Samuel F Passi; Amber N Stratman; Anastasia Sacharidou; M Patricia Revelo; Allie H Grossmann; Nikolaos A Diakos; George E Davis; Mark M Metzstein; Kevin J Whitehead; Dean Y Li Journal: J Clin Invest Date: 2011-04-01 Impact factor: 14.808
Authors: Romuald Girard; Maged D Fam; Hussein A Zeineddine; Huan Tan; Abdul Ghani Mikati; Changbin Shi; Michael Jesselson; Robert Shenkar; Meijing Wu; Ying Cao; Nicholas Hobson; Henrik B W Larsson; Gregory A Christoforidis; Issam A Awad Journal: J Neurosurg Date: 2016-08-05 Impact factor: 5.115
Authors: Christopher C Gibson; Weiquan Zhu; Chadwick T Davis; Jay A Bowman-Kirigin; Aubrey C Chan; Jing Ling; Ashley E Walker; Luca Goitre; Simona Delle Monache; Saverio Francesco Retta; Yan-Ting E Shiu; Allie H Grossmann; Kirk R Thomas; Anthony J Donato; Lisa A Lesniewski; Kevin J Whitehead; Dean Y Li Journal: Circulation Date: 2014-12-08 Impact factor: 29.690