Skip to main content
SpringerLink
Log in

Molecular Genetic Screening of CCM Patients: An Overview

  • Protocol
  • First Online:
Cerebral Cavernous Malformations (CCM)

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2152))

Abstract

Cerebral Cavernous Malformations (CCMs) are vascular lesions which can occur as a sporadic (80% of the cases) or a familial autosomal dominant disease (20%), the latter being characterized by the presence of multiple lesions. Three CCM genes have been identified in the last 10 years. More than 95% of familial cases and 60% of sporadic cases with multiple lesions harbor a germline heterozygous loss of function mutation in one of these 3 genes. Most mutations lead to a premature stop codon whatever the mechanism, including nonsense mutations, deletions, insertions and intronic mutations leading to abnormal splicing and frameshift. A combination of analyses, including sequencing and copy number analysis of germline DNA extracted from blood and cDNA analysis, are therefore required to ensure the best diagnostic sensitivity. Additional causative rare structural CCM gene anomalies have been identified in a research context, as well as rare causative missense mutations. These mutations are rarely searched for in a diagnostic context and explain part of the negative cases, in addition to germline mosaicism which occurs in some sporadic cases with multiple lesions. On top of germline mutations, somatic mutations occur on the wild-type allele in endothelial cells lining CCM lesions. These data established both the role of a double hit in the pathophysiology of CCM lesions and the heterogeneity of endothelial cells lining these lesions.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Labauge P, Denier C, Bergametti F et al (2007) Genetics of cavernous angiomas. Lancet Neurol 6:237–244

    Article  CAS  Google Scholar 

  2. Rigamonti D, Hadley MN, Drayer BP et al (1988) Cerebral cavernous malformations. Incidence and familial occurrence. N Engl J Med 319:343–347

    Article  CAS  Google Scholar 

  3. Labauge P, Laberge S, Brunereau L et al (1998) Hereditary cerebral cavernous angiomas: clinical and genetic features in 57 French families. Société Française de Neurochirurgie. Lancet 352:1892–1897

    Article  CAS  Google Scholar 

  4. Laberge-le Couteulx S, Jung HH, Labauge P et al (1999) Truncating mutations in CCM1, encoding KRIT1, cause hereditary cavernous angiomas. Nat Genet 23:189–193

    Article  CAS  Google Scholar 

  5. Liquori CL, Berg MJ, Siegel AM et al (2003) Mutations in a gene encoding a novel protein containing a phosphotyrosine-binding domain cause type 2 cerebral cavernous malformations. Am J Hum Genet 73:1459–1464

    Article  CAS  Google Scholar 

  6. Denier C, Labauge P, Brunereau L et al (2004) Clinical features of cerebral cavernous malformations patients with KRIT1 mutations. Ann Neurol 55:213–220

    Article  CAS  Google Scholar 

  7. Bergametti F, Denier C, Labauge P et al (2005) Mutations within the programmed cell death 10 gene cause cerebral cavernous malformations. Am J Hum Genet 76:42–51

    Article  CAS  Google Scholar 

  8. Craig HD, Günel M, Cepeda O et al (1998) Multilocus linkage identifies two new loci for a mendelian form of stroke, cerebral cavernous malformation, at 7p15-13 and 3q25.2-27. Hum Mol Genet 7:1851–1858

    Article  CAS  Google Scholar 

  9. Denier C, Labauge P, Bergametti F et al (2006) Genotype-phenotype correlations in cerebral cavernous malformations patients. Ann Neurol 60:550–556

    Article  Google Scholar 

  10. Spiegler S, Najm J, Liu J et al (2014) High mutation detection rates in cerebral cavernous malformation upon stringent inclusion criteria: one-third of probands are minors. Mol Genet Genomic Med 2:176–185

    Article  Google Scholar 

  11. Gunel M, Awad IA, Finberg K et al (1996) A founder mutation as a cause of cerebral cavernous malformation in Hispanic Americans. N Engl J Med 334:946–951

    Article  CAS  Google Scholar 

  12. Ortiz L, Costa AF, Bellido ML et al (2007) Study of cerebral cavernous malformation in Spain and Portugal: high prevalence of a 14 bp deletion in exon 5 of MGC4607 (CCM2 gene). J Neurol 254:322–326

    Article  Google Scholar 

  13. Cavé-Riant F, Denier C, Labauge P et al (2002) Spectrum and expression analysis of KRIT1 mutations in 121 consecutive and unrelated patients with cerebral cavernous malformations. Eur J Hum Genet 10:733–740

    Article  Google Scholar 

  14. Stahl S, Gaetzner S, Voss K et al (2008) Novel CCM1, CCM2, and CCM3 mutations in patients with cerebral cavernous malformations: in-frame deletion in CCM2 prevents formation of a CCM1/CCM2/CCM3 protein complex. Hum Mutat 29:709–717

    Article  CAS  Google Scholar 

  15. Spiegler S, Rath M, Paperlein C et al (2018) Cerebral cavernous malformations: an update on prevalence, molecular genetic analyses, and genetic counselling. Mol Syndromol 9:60–69

    Article  CAS  Google Scholar 

  16. Riant F, Odent S, Cecillon M et al (2014) Deep intronic KRIT1 mutation in a family with clinically silent multiple cerebral cavernous malformations. Clin Genet 86:585–588

    Article  CAS  Google Scholar 

  17. Rath M, Jenssen SE, Schwefel K et al (2017) High-throughput sequencing of the entire genomic regions of CCM1/KRIT1, CCM2 and CCM3/PDCD10 to search for pathogenic deep-intronic splice mutations in cerebral cavernous malformations. Eur J Med Genet 60:479–484

    Article  Google Scholar 

  18. Spiegler S, Rath M, Hoffjan S et al (2018) First large genomic inversion in familial cerebral cavernous malformation identified by whole genome sequencing. Neurogenetics 19:55–59

    Article  CAS  Google Scholar 

  19. Zawistowski JS, Stalheim L, Uhlik MT et al (2005) CCM1 and CCM2 protein interactions in cell signaling: implications for cerebral cavernous malformations pathogenesis. Hum Mol Genet 14:2521–2531

    Article  CAS  Google Scholar 

  20. Fisher OS, Liu W, Zhang R et al (2015) Structural basis for the disruption of the cerebral cavernous malformations 2 (CCM2) interaction with Krev interaction trapped 1 (KRIT1) by disease-associated mutations. J Biol Chem 290:2842–2853

    Article  CAS  Google Scholar 

  21. Draheim KM, Li X, Zhang R et al (2015) CCM2-CCM3 interaction stabilizes their protein expression and permits endothelial network formation. J Cell Biol 208:987–1001

    Article  CAS  Google Scholar 

  22. Rath M, Spiegler S, Nath N et al (2017) Constitutional de novo and postzygotic mutations in isolated cases of cerebral cavernous malformations. Mol Genet Genomic Med 5:21–27

    Article  CAS  Google Scholar 

  23. Gianfrancesco F, Esposito T, Penco S et al (2008) ZPLD1 gene is disrupted in a patient with balanced translocation that exhibits cerebral cavernous malformations. Neuroscience 155:345–349

    Article  CAS  Google Scholar 

  24. Laviña B, Castro M, Niaudet C et al (2018) Defective endothelial cell migration in the absence of Cdc42 leads to capillary-venous malformations. Development 145:dev161182

    Article  Google Scholar 

  25. Castro M, Laviña B, Ando K et al (2019) CDC42 deletion elicits cerebral vascular malformations via increased MEKK3-dependent KLF4 expression. Circ Res 124:1240–1252

    Article  CAS  Google Scholar 

  26. Riant F, Bergametti F, Fournier H-D et al (2013) CCM3 mutations are associated with early-onset cerebral hemorrhage and multiple meningiomas. Mol Syndromol 4:165–172

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Gault J, Shenkar R, Recksiek P et al (2005) Biallelic somatic and germ line CCM1 truncating mutations in a cerebral cavernous malformation lesion. Stroke 36:872–874

    Article  Google Scholar 

  28. Akers AL, Johnson E, Steinberg GK et al (2009) Biallelic somatic and germline mutations in cerebral cavernous malformations (CCMs): evidence for a two-hit mechanism of CCM pathogenesis. Hum Mol Genet 18:919–930

    Article  CAS  Google Scholar 

  29. Pagenstecher A, Stahl S, Sure U et al (2009) A two-hit mechanism causes cerebral cavernous malformations: complete inactivation of CCM1, CCM2 or CCM3 in affected endothelial cells. Hum Mol Genet 18:911–918

    Article  CAS  Google Scholar 

  30. McDonald DA, Shi C, Shenkar R et al (2014) Lesions from patients with sporadic cerebral cavernous malformations harbor somatic mutations in the CCM genes: evidence for a common biochemical pathway for CCM pathogenesis. Hum Mol Genet 23:4357–4370

    Article  CAS  Google Scholar 

  31. Detter MR, Snellings DA, Marchuk DA (2018) Cerebral cavernous malformations develop through clonal expansion of mutant endothelial cells. Circ Res 123:1143–1151

    Article  CAS  Google Scholar 

  32. Malinverno M, Maderna C, Abu Taha A et al (2019) Endothelial cell clonal expansion in the development of cerebral cavernous malformations. Nat Commun 10:2761

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. INSERM U1141, Assistance Publique Hôpitaux de Paris, Paris University, Paris, France

    Elisabeth Tournier-Lasserve

Authors
  1. Elisabeth Tournier-Lasserve
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Elisabeth Tournier-Lasserve .

Editor information

Editors and Affiliations

  1. Department of Biotechnology, Chemistry and Pharmacy, University of Siena, SIENA, Italy

    Lorenza Trabalzini

  2. Department of Biotechnology, Chemistry and Pharmacy, University of Siena, SIENA, Italy

    Federica Finetti

  3. Department of Clinical and Biological Sciences, School of Medicine and Surgery, University of Torino, ORBASSANO (TORINO), Italy

    Saverio Francesco Retta

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Tournier-Lasserve, E. (2020). Molecular Genetic Screening of CCM Patients: An Overview. In: Trabalzini, L., Finetti, F., Retta, S. (eds) Cerebral Cavernous Malformations (CCM) . Methods in Molecular Biology, vol 2152. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0640-7_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0640-7_4

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0639-1

  • Online ISBN: 978-1-0716-0640-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation