MLPA-analysis of the CCM genes in patients with various forms of Cerebral Cavernous Malformations of the CNS in Russian Federation

Mutations in the CCM genes causing the formation of cavernous malformations (СМ) are being actively studied and their list is constantly growing. Mutations in the CCM genes are usually analyzed using two main approaches: multiplex ligation-dependent probe amplification (MLPA) - to search for large deletions/insertions and exon sequencing - to search for mutations leading to amino acid substitutions, the appearance of premature translation termination, reading frame shifts and splice site junction. On the basis of these data, cheaper diagnostic test systems are created that allow the identification of mutant alleles in order to predict the possible occurrence and development of cerebral cavities. Objective. Identification of large deletions in CCM genes associated with the developing of cerebral СМ in patients with sporadic and hereditary forms of the disease in the Russian population. Methods. Blood samples from 92 selected patients were examined, among them - 45 with a hereditary form of the disease, 2 - with clinically confirmed familial cases and 45 - so-called sporadic cases, as well as in 10 healthy relatives. Presence of large deletions/duplications was detected by multiplex ligation-dependent probe amplification (MPLA). Results. Major rearrangements in one of the CCM genes ( CCM1 , CCM2 , CCM3 ) were identified in 10 samples, including 2 patients with conditionally familial form of CM, and 3 patients with a sporadic form. Of the seven types of mutations identified, four were not previously described. Two deletions in the CCM3 [PDCD10] gene were identified, for which such mutations have not yet been described. The ratio of mutations in the CCM1 , CCM2 and CCM3 genes was 40%, 30% and 30%, respectively. The most aggressive clinical course was observed in patients with mutations in the CCM3 gene. Conclusion. The analysis showed that large deletions in the genes of CCM are quite common in patients with sporadic and hereditary forms of the disease in the Russian population and accounted for approximately 11% of the studied sample. At the same time, new types of mutations that are not described in other populations have been identified

кавернома, кавернозная мальформация ЦНС, наследственные каверномы, мутации генов каверном, MLPA, familial cerebral cavernous malformation, genes CCM1, CCM2, CCM3, detection of gene mutations

1. Hayman LA, Evans RA, Ferrell RE, Fahr LM, Ostrow P, Riccardi VM. Familial cavernous angiomas: natural history and genetic study over a 5-year period. Am J Med Genet. 1982 Feb;11(2):147-60. doi: https://doi.org/10.1002/ajmg.1320110205

2. Bicknell JM, Carlow TJ, Kornfeld M, Stovring J, Turner P. Familial cavernous angiomas. Arch Neurol. 1978 Nov;35(11):746-9. doi: https://doi.org/10.1001/archneur.1978.00500350050010

3. Sahoo T, Johnson EW, Thomas JW, Kuehl PM, Jones TL, Dokken CG, Touchman JW, Gallione CJ, Lee-Lin SQ, Kosofsky B, Kurth JH, Louis DN, Mettler G, Morrison L, Gil-Nagel A, Rich SS, Zabramski JM, Boguski MS, Green ED, Marchuk DA. Mutations in the gene encoding KRIT1, a Krev-1/rap1a binding protein, cause cerebral cavernous malformations (CCM1). Hum Mol Genet. 1999 Nov;8(12):2325-33. doi: https://doi.org/10.1093/hmg/8.12.2325

4. Liquori CL, Berg MJ, Siegel AM, Huang E, Zawistowski JS, Stoffer T, Verlaan D, Balogun F, Hughes L, Leedom TP, Plummer NW, Cannella M, Maglione V, Squitieri F, Johnson EW, Rouleau GA, Ptacek L, Marchuk DA. Mutations in a gene encoding a novel protein containing a phosphotyrosine-binding domain cause type 2 cerebral cavernous malformations. Am J Hum Genet. 2003 Dec;73(6):1459-64. Epub 2003 Nov 17. doi: https://doi.org/10.1086/380314

5. Denier C, Goutagny S, Labauge P, Krivosic V, Arnoult M, Cousin A, Benabid AL, Comoy J, Frerebeau P, Gilbert B, Houtteville JP, Jan M, Lapierre F, Loiseau H, Menei P, Mercier P, Moreau JJ, Nivelon-Chevallier A, Parker F, Redondo AM, Scarabin JM, Tremoulet M, Zerah M, Maciazek J, Tournier-Lasserve E; Sociеtе Franсaise de Neurochirurgie. Mutations within the MGC4607 gene cause cerebral cavernous malformations. Am J Hum Genet. 2004 Feb;74(2):326-37. Epub 2004 Jan 22. doi: https://doi.org/10.1086/381718

6. Bergametti F, Denier C, Labauge P, Arnoult M, Boetto S, Clanet M, Coubes P, Echenne B, Ibrahim R, Irthum B, Jacquet G, Lonjon M, Moreau JJ, Neau JP, Parker F, Tremoulet M, Tournier-Lasserve E; Sociеtе Franсaise de Neurochirurgie. Mutations within the programmed cell death 10 gene cause cerebral cavernous malformations. Am J Hum Genet. 2005 Jan;76(1):42-51. Epub 2004 Nov 12. doi: https://doi.org/10.1086/426952

7. Penco S, Ratti R, Bianchi E, Citterio A, Patrosso MC, Marocchi A, Tassi L, La Camera A, Collice M. Molecular screening test in familial forms of cerebral cavernous malformation: the impact of the Multiplex Ligation-dependent Probe Amplification approach. Neurosurg. 2009 May;110(5):929-34. doi: https://doi.org/10.3171/2008.8.17640.

8. Liquori CL, Berg MJ, Squitieri F, Leedom TP, Ptacek L, Johnson EW, Marchuk DA. Deletions in CCM2 are common cause of cerebral cavernous malformations. Am J Hum Genet. 2007 Jan;80(1):69-75. Epub 2006 Nov 14. doi: https://doi.org/10.1086/510439

9. D’Angelo R1, Marini V, Rinaldi C, Origone P, Dorcaratto A, Avolio M, Goitre L, Forni M, Capra V, Alafaci C, Mareni C, Garrе C, Bramanti P, Sidoti A, Retta SF, Amato A. Mutation analysis of CCM1, CCM2 and CCM3 genes in a cohort of Italian patients with cerebral cavernous malformation. Brain Pathol. 2011 Mar;21(2):215-24. Epub 2010 Oct 4. doi: https://doi.org/10.1111/j.1750-3639.2010.00441.x

10. Mondjar R., Solano F., Rubio R., Delgado M., Perez-Sempere A., Gonzalez-Meneses A., Vendrell T., Izquierdo G., Martinez-Mir A., Lucas M. Mutation Prevalence of Cerebral Cavernous Malformation Genes in Spanish Patients. PloS one 2014, 9; e86286. PLoS One. 2014 Jan 23;9(1):e86286. eCollection 2014. doi: https://10.1371/journal.pone.0086286

11. Боринская СА, Хуснутдинова ЭК. Этногеномика: история с географией. Человек. 2002; 1:19-30.

12. Лимборская СА, Хуснутдинова ЭК, Балановская ЕВ. Этногеномика и геногеография народов Восточной Европы. М.: Наука, 2002.

13. Maiuri F, Cappabianca P, Gangemi M, De Caro Mdel B, Esposito F, Pettinato G, de Divitiis O, Mignogna C, Strazzullo V, de Divitiis E. Clinical progression and familial occurrence of cerebral cavernous angiomas: the role of angiogenic and growth factors. Neurosurg Focus. 2006 Jul15;21(1):e3. doi: http://dx.doi.org/10.3171/foc.2006.21.1.4

14. Gaetzner S, Stahl S, Surucu O, Schaafhausen A, Halliger-Keller B, Bertalanffy H, Sure U, Felbor U. CCM1 gene deletion identified by MLPA in cerebral cavernous malformation. Neurosurg Rev. 2007 Apr;30(2):155-9; discussion 159-60. Epub 2006 Dec 23. doi: https://doi.org/10.1007/s10143-006-0057-1

15. Felbor U, Gaetzner S, Verlaan DJ, Vijzelaar R, Rouleau GA, Siegel AM. Large germline deletions and duplication in isolated cerebral cavernous malformation patients. Neurogenetics. 2007 Apr;8(2):149-53. Epub 2007 Jan 9. doi: https://doi.org/10.1007/s10048-006-0076-7

16. Мационис АЭ, Петров АВ, Горелик МЗ, Завалишина ЛЭ. Исследование численных нарушений генов при раке молочной железы методом мультиплексной лигазозависимой амплификации зондов. Архив патологии. 2014;76(4):15-17.

17. Гусина АА, Мясников СО, Гусина НБ. Метод мультиплексной амплификации лигированных зондов в диагностике синдрома Марфана. Медицинская генетика. 2018; 17 (4): 37-41. doi: https://doi.org/10.25557/2073-7998.2018.04.37-41

18. Lai KK, Lo IF, Tong TM, Cheng LY, Lam ST. Detecting exon deletions and duplications of the DMD gene using Multiplex Ligation-dependent Probe Amplification (MLPA). Clin Biochem. 2006 Apr;39(4):367-72. Epub 2006 Jan 17. doi: https://doi.org/10.1016/j.clinbiochem.2005.11.019

19. Denier C, Labauge P, Bergametti F, Marchelli F, Riant F, Arnoult M, Maciazek J, Vicaut E, Brunereau L, Tournier-Lasserve E; Sociеtе Franсaise de Neurochirurgie. Genotype-phenotype correlations in cerebral cavernous malformations patients. Ann Neurol. 2006 Nov;60(5):550-6. doi: https://doi.org/10.1002/ana.20947

20. Maiuri F, Cappabianca P, Gangemi M, De Caro Mdel B, Esposito F, Pettinato G, de Divitiis O, Mignogna C, Strazzullo V, de Divitiis E. Clinical progression and familial occurrence of cerebral cavernous angiomas: the role of angiogenic and growth factors. Neurosurg Focus. 2006 Jul 15;21(1):e3. doi: https://doi.org/10.3171/foc.2006.21.1.4

21. Nikoubashman O, Wiesmann M, Tournier-Lasserve E, Mankad K, Bourgeois M, Brunelle F, Sainte-Rose C, Wiesmann M, Zerah M, Di Rocco F. Natural history of cerebral dot-like cavernomas. Clin Radiol. 2013 Aug;68(8):e453-9. Epub 2013 May 8. doi: https://doi.org/10.1016/j.crad.2013.02.010