medgenМедицинская генетикаMedical Genetics2073-7998Publishing House «Genius Media» LLC10.25557/2073-7998.2018.12.36-43medgen-610Research ArticleОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯORIGINAL RESEARCHНонсенс-мутация GLN1233* и заменa ARG326GLN в гене MYBPC3 у пациентов с гипертрофической кардиомиопатией в БеларусиGLN1233* nonsens-mutation and ARG326GLN polymorphism of MYBPC3 gene in patients with hypertrophic cardiomyopathy in BelarusЧаковаН. Н.ChakovaN. N.n.chakova@igc.byНиязоваС. С.NiyazovaS. S.noemail@neicon.ruКомиссароваС. М.KomissarovaS. M.noemail@neicon.ruСасиновичМ. А.SasinovichM. A.noemail@neicon.ruГончаренкоМ. Г.GoncharenkoM. G.noemail@neicon.ruГНУ «Институт генетики и цитологии Национальной Академии Наук Беларуси»РоссияThe Institute of Genetics and Cytology of the National Academy of Sciences of BelarusRussian FederationРеспубликанский научно-практический центр «Кардиология»РоссияRepublican Research and Practical Centre «Cardiology»Russian Federation20180112201817123643Copyright © Чакова Н.Н., Ниязова С.С., Комиссарова С.М., Сасинович М.А., Гончаренко М.Г., 20182018Чакова Н.Н., Ниязова С.С., Комиссарова С.М., Сасинович М.А., Гончаренко М.Г.Chakova N.N., Niyazova S.S., Komissarova S.M., Sasinovich M.A., Goncharenko M.G.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.medgen-journal.ru/jour/article/view/610

Актуальность . Мутация р.Gln1233* (rs397516037) и замена р.Arg326Gln (rs34580776) в гене MYBPC 3 выявлены с различной частотой встречаемости у пациентов с гипертрофической кардиомиопатией (ГКМП) во многих популяциях. Опубликованные данные по этим генетическим изменениям различаются по интерпретации их патогенности. Определен эффект основателя для р.Gln1233* в некоторых популяциях. Целью данного исследования являлись оценка распространенности замен rs397516037 и rs34580776 в гене MYBPC 3 у белорусских пациентов с ГКМП и у лиц контрольной группы, проверка гипотезы об эффекте основателя для мутации р.Gln1233* , а также описание клинических проявлений заболевания у пациентов с этой мутацией. Материалы и методы . У 85 пациентов генетический анализ проводили методом NGS. У 250 неродственных пациентов, 13 родственников пробандов с мутацией р.Gln1233* и 127 лиц контрольной группы проводили направленный поиск замены р.Arg326Gln методом ПЦР-ПДРФ анализа. Выявление мутации р.Gln1233* осуществляли методом автоматического секвенирования у всех носителей р.Arg326Gln , а также у 113 пациентов без этой замены. У обладателей хотя бы одной замены генотипировали локусы р.Val849Val (c.2547C>T, rs3729953) и р.Glu1096Glu (c.3288G>A, rs1052373) методом ПЦР-ПДРФ анализа для определения гаплотипа. Результаты . У 5,37% (18 из 335) белорусских пациентов с ГКМП выявлены нонсенс-мутация р.Gln1233* и замена р.Arg326Gln в цис-положении. Отсутствие мутации р.Gln1233* в контрольной группе подтверждает ее диагностическую значимость в отношении развития ГКМП. Замена р.Arg326Gln без мутации р.Gln1233* обнаружена у 5,07% (17 из 335) пациентов и у 3,94% лиц контрольной выборки, что указывает на незначимость данного полиморфизма в развитии заболевания. Установлен эффект основателя для мутации р.Gln1233* на территории Беларуси. Дана клиническая характеристика ГКМП у пробандов и их ближайших родственников с этой мутацией. Выводы . Нонсенс-мутация р.Gln1233* (rs397516037) является самой частой среди обнаруженных мутаций у белорусских пациентов с ГКМП. Течение заболевания у носителей мутации p.Gln1233* зависело от возраста и варьировало от малосимптомного у более молодых пациентов (18-40 лет) до тяжелого течения с неблагоприятным прогнозом вплоть до летального исхода вследствие прогрессирования хронической сердечной недостаточности (после 40 лет).

Relevance . In many populations of patients with hypertrophic cardiomyopathy (HCM), mutation of р.Gln1233* (rs397516037) and р.Arg326Gln (rs34580776) substitution in MYBPC 3 have been identified with different frequency. The published data of these genetic changes differ in the interpretation of their pathogenicity. The founder effect has been described for р.Gln1233* for some populations. The aim was to assess the incidence of rs397516037 and rs34580776 substitution in MYBPC 3 in Belarusian patients with HCM and in control group; to test the hypothesis about the effects of the founder for р.Gln1233* mutation, and to describe clinical features of the disease in the group of patients with this mutation. Materials and methods . Genetic analysis was made by NGS for 85 individuals. A guided search for a р.Arg326Gln substitution by PCR-RFLP was made in 250 non-related individuals, 13 proband relatives with р.Gln1233* mutations and 127 controls. The р.Gln1233* mutation identification was made using automated sequencing method in all carriers of р.Arg326Gln , and in 113 individuals without the substitution. In those carriers with at least one substitution, р.Val849Val (c.2547C>T, rs3729953) and р.Glu1096Glu (c.3288G>A, rs1052373) loci were genotyped by PCR-RFLP to identify the haplotype. Results . In 5.37% (18 of 335) Belarusian individuals with HCM, р.Gln1233* nonsense-mutation and р.Arg326Gln substitution in cis-arrangement were established. No р.Gln1233* mutation in the controls suggested its diagnostic significance concerning the development of HCM. The р.Arg326Gln substitution without present р.Gln1233* mutation was established in 5.07% (17 of 335) individuals and in 3.94% controls of the sample suggesting the insignificance of this polymorphism in the development of the condition. The effect of the founder for р.Gln1233* mutation for Belarus has been established. Clinical characteristics of HCM has been proposed for probands and closest relatives who have this mutation. Conclusions . р.Gln1233* nonsense-mutation (rs397516037) is the most frequent mutation among mutations identified in Belarusian individuals diagnosed with HCM. The course of the disease in p.Gln1233* carriers varied from mildly symptomatic at a younger age (18 to 40 y.o.) to severe with malignant prognosis including fatal outcome due to CHF progression.

гипертрофическая кардиомиопатиямутация р.Gln1233* и замена р.Arg326Gln в гене MYBPC3гаплотипэффект основателяклинические проявленияhypertrophic cardiomyopathyр.Gln1233* mutation and р.Arg326Gln substitution in MYBPC3haplotypefounder effectclinical featuresReferencesCarrier L, Bonne G, Bahrend E et al. Organization and sequence of human cardiac myosin binding protein C gene (MYBPC3)and identification of mutations predicted to produce truncated proteins in familial hypertrophic cardiomyopathy. Circ Res. 1997;80:427-434.Carrier L, Bonne G, Bahrend E et al. Organization and sequence of human cardiac myosin binding protein C gene (MYBPC3)and identification of mutations predicted to produce truncated proteins in familial hypertrophic cardiomyopathy. Circ Res. 1997;80:427-434.Richard P, Charron P, Carrier L et al. Hypertrophic cardiomyopathy: distribution of disease genes, spectrum of mutations, and implications for a molecular diagnosis strategy. Circulation. 2003;107(17):2227-2232.Richard P, Charron P, Carrier L et al. Hypertrophic cardiomyopathy: distribution of disease genes, spectrum of mutations, and implications for a molecular diagnosis strategy. Circulation. 2003;107(17):2227-2232.Curila K, Benesova L, Penicka M et al. Spectrum and clinical manifestations of mutations in genes responsible for hypertrophic cardiomyopathy. Acta Cardiol. 2012 Feb;67(1):23-29.Curila K, Benesova L, Penicka M et al. Spectrum and clinical manifestations of mutations in genes responsible for hypertrophic cardiomyopathy. Acta Cardiol. 2012 Feb;67(1):23-29.Niimura H, Patton KK, McKenna WJ et al. Sarcomere protein gene mutations in hypertrophic cardiomyopathy of the elderly. Circulation. 2002 Jan 29;105(4):446-451.Niimura H, Patton KK, McKenna WJ et al. Sarcomere protein gene mutations in hypertrophic cardiomyopathy of the elderly. Circulation. 2002 Jan 29;105(4):446-451.Поляк МЕ, Ховалыг АБ, Букаева АА и др. Спектр мутаций в гене MYBPC3 у пациентов с гипертрофической кардиомиопатией. Медицинская генетика. 2016;15(8): 26-29.Поляк МЕ, Ховалыг АБ, Букаева АА и др. Спектр мутаций в гене MYBPC3 у пациентов с гипертрофической кардиомиопатией. Медицинская генетика. 2016;15(8): 26-29.Ingles J, Doolan A, Chiu C et al. Compound and double mutations in patients with hypertrophic cardiomyopathy: implications for genetic testing and counselling. J Med Genet. 2005 Oct;42(10):e59.Ingles J, Doolan A, Chiu C et al. Compound and double mutations in patients with hypertrophic cardiomyopathy: implications for genetic testing and counselling. J Med Genet. 2005 Oct;42(10):e59.Roncarati R, Latronico MV, Musumeci B et al. Unexpectedly low mutation rates in beta-myosin heavy chain and cardiac myosin binding protein genes in Italian patients with hypertrophic cardiomyopathy. J Cell Physiol. 2011 Nov;226(11):2894-2900.Roncarati R, Latronico MV, Musumeci B et al. Unexpectedly low mutation rates in beta-myosin heavy chain and cardiac myosin binding protein genes in Italian patients with hypertrophic cardiomyopathy. J Cell Physiol. 2011 Nov;226(11):2894-2900.Erdmann J, Raible J, Maki-Abadi J et al. Spectrum of clinical phenotypes and gene variants in cardiac myosin-binding protein C mutation carriers with hypertrophic cardiomyopathy. J Am Coll Cardiol. 2001 Aug;38(2):322-330.Erdmann J, Raible J, Maki-Abadi J et al. Spectrum of clinical phenotypes and gene variants in cardiac myosin-binding protein C mutation carriers with hypertrophic cardiomyopathy. J Am Coll Cardiol. 2001 Aug;38(2):322-330.Bos JM, Will ML, Gersh BJ et al. Characterization of a phenotype-based genetic test prediction score for unrelated patients with hypertrophic cardiomyopathy. Mayo Clin Proc. 2014 Jun;89(6):727-737.Bos JM, Will ML, Gersh BJ et al. Characterization of a phenotype-based genetic test prediction score for unrelated patients with hypertrophic cardiomyopathy. Mayo Clin Proc. 2014 Jun;89(6):727-737.Tоth T, Nagy V, Faludi R et al. The Gln1233ter mutation of the myosin binding protein C gene: causative mutation or innocent polymorphism in patients with hypertrophic cardiomyopathy. Int J Cardiol. 2011 Dec 1;153(2):216-219.Tоth T, Nagy V, Faludi R et al. The Gln1233ter mutation of the myosin binding protein C gene: causative mutation or innocent polymorphism in patients with hypertrophic cardiomyopathy. Int J Cardiol. 2011 Dec 1;153(2):216-219.Mathew CC. The isolation of high molecular weight eukaryotic DNA. Methods Mol Biol. 1984;2:31-34.Mathew CC. The isolation of high molecular weight eukaryotic DNA. Methods Mol Biol. 1984;2:31-34.Glotov AS, Kazakov SV, Zhukova EA et al. Targeted next-generation sequencing (NGS) of nine candidate genes with custom AmpliSeq in patients and a cardiomyopathy risk group. Clin Chim Acta. 2015;446:132-140.Glotov AS, Kazakov SV, Zhukova EA et al. Targeted next-generation sequencing (NGS) of nine candidate genes with custom AmpliSeq in patients and a cardiomyopathy risk group. Clin Chim Acta. 2015;446:132-140.Rafael JF, Cruz FEDS Filho, Carvalho ACC et al. Myosin-binding Protein C Compound Heterozygous Variant Effect on the Phenotypic Expression of Hypertrophic Cardiomyopathy. Arq Bras Cardiol. 2017 Apr; 108(4): 354-360.Rafael JF, Cruz FEDS Filho, Carvalho ACC et al. Myosin-binding Protein C Compound Heterozygous Variant Effect on the Phenotypic Expression of Hypertrophic Cardiomyopathy. Arq Bras Cardiol. 2017 Apr; 108(4): 354-360.Maron BJ, Niimura H, Casey SA et al. Development of left ventricular hypertrophy in adults in hypertrophic cardiomyopathy caused by cardiac myosin-binding protein C gene mutations. J Am Coll Cardiol. 2001 Aug;38(2):315-321.Maron BJ, Niimura H, Casey SA et al. Development of left ventricular hypertrophy in adults in hypertrophic cardiomyopathy caused by cardiac myosin-binding protein C gene mutations. J Am Coll Cardiol. 2001 Aug;38(2):315-321.Jааskelаinen P, Kuusisto J, Miettinen R et al. Mutations in the cardiac myosin-binding protein C gene are the predominant cause of familial hypertrophic cardiomyopathy in eastern Finland. J Mol Med. 2002 Jul;80(7):412-422.Jааskelаinen P, Kuusisto J, Miettinen R et al. Mutations in the cardiac myosin-binding protein C gene are the predominant cause of familial hypertrophic cardiomyopathy in eastern Finland. J Mol Med. 2002 Jul;80(7):412-422.Jааskelаinen P, Heliо T, Aalto-Setаlа K et al. A new common mutation in the cardiac beta-myosin heavy chain gene in Finnish patients with hypertrophic cardiomyopathy. Ann Med. 2014 Sep;46(6):424-429.Jааskelаinen P, Heliо T, Aalto-Setаlа K et al. A new common mutation in the cardiac beta-myosin heavy chain gene in Finnish patients with hypertrophic cardiomyopathy. Ann Med. 2014 Sep;46(6):424-429.Lopes LR, Zekavati A, Syrris P et al. Genetic complexity in hypertrophic cardiomyopathy revealed by high-throughput sequencing. J Med Genet. 2013 Apr;50(4): 228-239.Lopes LR, Zekavati A, Syrris P et al. Genetic complexity in hypertrophic cardiomyopathy revealed by high-throughput sequencing. J Med Genet. 2013 Apr;50(4): 228-239.Brito D, Miltenberger-Miltenyi G, Vale Pereira S et al. Sarcomeric hypertrophic cardiomyopathy: genetic profile in a Portuguese population. Rev Port Cardiol. 2012 Sep;31(9):577-587.Brito D, Miltenberger-Miltenyi G, Vale Pereira S et al. Sarcomeric hypertrophic cardiomyopathy: genetic profile in a Portuguese population. Rev Port Cardiol. 2012 Sep;31(9):577-587.Van Driest SL, Jaeger MA, Ommen SR et al. Comprehensive analysis of the beta-myosin heavy chain gene in 389 unrelated patients with hypertrophic cardiomyopathy. J Am Coll Cardiol. 2004;44(3): 602-610.Van Driest SL, Jaeger MA, Ommen SR et al. Comprehensive analysis of the beta-myosin heavy chain gene in 389 unrelated patients with hypertrophic cardiomyopathy. J Am Coll Cardiol. 2004;44(3): 602-610.Lopes LR, Syrris P, Guttmann OP et al. Novel genotype-phenotype associations demonstrated by high-throughput sequencing in patients with hypertrophic cardiomyopathy. Heart. 2015 Feb;101(4):294-301.Lopes LR, Syrris P, Guttmann OP et al. Novel genotype-phenotype associations demonstrated by high-throughput sequencing in patients with hypertrophic cardiomyopathy. Heart. 2015 Feb;101(4):294-301.Ehlermann P, Weichenhan D, Zehelein J et al. Adverse events in families with hypertrophic or dilated cardiomyopathy and mutations in the MYBPC3 gene. BMC Med Genet. 2008 Oct 28;9:95.Ehlermann P, Weichenhan D, Zehelein J et al. Adverse events in families with hypertrophic or dilated cardiomyopathy and mutations in the MYBPC3 gene. BMC Med Genet. 2008 Oct 28;9:95.Fokstuen S, Lyle R, Munoz A et al. A DNA resequencing array for pathogenic mutation detection in hypertrophic cardiomyopathy. Hum Mutat. 2008 Jun;29(6):879-885.Fokstuen S, Lyle R, Munoz A et al. A DNA resequencing array for pathogenic mutation detection in hypertrophic cardiomyopathy. Hum Mutat. 2008 Jun;29(6):879-885.

The authors declare that there are no conflicts of interest present.