二代测序技术筛查肥厚型心肌病伴房颤家系致病基因研究
|
摘要
目的对1个肥厚型心肌病伴房颤先证者及其家系成员进行致病基因筛查研究。方法收集先证者及其家系成员外周血,提取基因组DNA,应用二代测序法对先证者进行心血管疾病候选基因突变检测。发现可疑致病位点后,进一步通过Sanger测序法在家系内其他成员以及100个健康个体者中进行验证。用PolyPhen-2等生物信息学软件对新突变位点进行致病突变分析。家系资料收集包括临床表现、体格检查、心电图、超声心动图。结果先证者及家系内多个成员均携带PRDM16基因c.3124G>A(p.G1042R)杂合突变,但先证者父母及100例健康个体中并未发现该突变,因此,该突变可能为新生突变(生殖细胞嵌合)。生物信息学分析结果显示PRDM16基因c.3124G>A(p.G1042R)杂合突变位于保守位点,可能影响蛋白功能,为致病性突变。结论本研究发现1个肥厚型心肌病伴房颤的家系中存在PRDM16基因c.3124G>A(p.G1042R)杂合突变,该突变所在区域在不同种系间高度保守,可能是该肥厚型心肌病家系的致病基因突变位点。
Objective To identify apotential pathogenic gene mutation among a family with hypertrophic cardiomyopathy(HCM)and atrial fibrillation(AF).Methods Clinical data,family history,ECG,and echocardiogram were collected from the proband as well as family members.Genomic DNA was extracted from blood sample,and a gene panel related to hereditary cardiovascular diseases was detected using next-generation sequencing.Mutation identified in the proband was confirmed by Sanger sequencing in other family members and100 healthy controls.The potential pathogenicity of the identified mutation was assessed by PolyPhen-2,SIFT and Mutation Taster software.Results Three patients showed clinical symptoms related to the pathology.A novel PRDM16 gene c.3124 G>A(p.G1042 R)mutation was identified in the proband and several members in this family.However,this mutation was not found in the proband's parent and the healthy controls,so it could be a de novo mutation.This mutation was located in a high evolutionary conservation area among different species.Both conservatism and bioinformatics results indicated that the mutation probably had affected the function of the protein.Conclusion The novel missense mutation of PRDM16 gene c.3124 G> A might be the disease-causing gene mutation in this Chinese pedigree with familiar HCM and AF.
Objective To identify apotential pathogenic gene mutation among a family with hypertrophic cardiomyopathy(HCM)and atrial fibrillation(AF).Methods Clinical data,family history,ECG,and echocardiogram were collected from the proband as well as family members.Genomic DNA was extracted from blood sample,and a gene panel related to hereditary cardiovascular diseases was detected using next-generation sequencing.Mutation identified in the proband was confirmed by Sanger sequencing in other family members and100 healthy controls.The potential pathogenicity of the identified mutation was assessed by PolyPhen-2,SIFT and Mutation Taster software.Results Three patients showed clinical symptoms related to the pathology.A novel PRDM16 gene c.3124 G>A(p.G1042 R)mutation was identified in the proband and several members in this family.However,this mutation was not found in the proband's parent and the healthy controls,so it could be a de novo mutation.This mutation was located in a high evolutionary conservation area among different species.Both conservatism and bioinformatics results indicated that the mutation probably had affected the function of the protein.Conclusion The novel missense mutation of PRDM16 gene c.3124 G> A might be the disease-causing gene mutation in this Chinese pedigree with familiar HCM and AF.
引文
[1]VESELKA J,ANAVEKAR NS,CHARRON P.Hypertrophic obstructive cardiomyopathy[J].Lancet,2017,389(10075):1253-1267.
[2]MARON BJ,MARON MS.Hypertrophic cardiomyopathy[J].Lancet,2013,381(9862):242-255.
[3]ELLIOTT PM,ANASTASAKIS A,BORGER MA,et al.2014ESC Guidelines on Diagnosis and Management of Hypertrophic Cardiomyopathy:the Task Force for the Diagnosis and Management of Hypertrophic Cardiomyopathy of the European Society of Cardiology(ESC)[J].Eur Heart J,2014,35(39):2733-2779.
[4]KRAKER J,VISWANATHAN SK,KNOLL R,et al.Recent advances in the molecular genetics of familial hypertrophic cardiomyopathy in south Asian descendants[J].Front Physiol,2016,7:499.
[5]FU L,LUO S,CAI S,et al.Identification of LAMP2 mutations in early-onset danon disease with hypertrophic cardiomyopathy by targeted next-generation sequencing[J].Am J Cardiol,2016,118(6):888-894.
[6]MANGO R,LUCHETTI A,SANGIUOLO R,et al.Next generation sequencing and linkage analysis for the molecular diagnosis of a novel overlapping syndrome characterized by hypertrophic cardiomyopathy and typical electrical instability of Brugada syndrome[J].Circ J,2016,80(4):938-949.
[7]WEISSLER-SNIR A,CREAN A,RAKOWSKI H.The role of imaging in the diagnosis and management of hypertrophic cardiomyopathy[J].Expert Rev Cardiovasc Ther,2016,14(1):51-74.
[8]杨雪蓉,陈代文,余冰,等.PRDM16的研究进展[J].动物营养学报,2010,22(6):1477-481.
[9]PINHEIRO I,MARGUERON R,SHUKEIR N,et al.Prdm3and Prdm16 are H3K9me1 methyltransferases required for mammalian heterochromatin integrity[J].Cell,2012,150(5):948-960.
[10]BJORK BC,TURBE-DOAN A,PRYSAK M,et al.Prdm16is required for normal palatogenesis in mice[J].Hum Mol Genet,2010,19(5):774-789.
[11]ARNDT AK,SCHAFER S,DRENCKHAHN JD,et al.Fine mapping of the 1p36 deletion syndrome identifies mutation of PRDM16 as a cause of cardiomyopathy[J].Am J Hum Genet,2013,93(1):67-77.
[12]OLIVOTTO I,CECCHI F,CASEY SA,et al.Impact of atrial fibrillation on the clinical course of hypertrophic cardiomyopathy[J].Circulation,2001,104(21):2517-2524.
[13]伍熙崔,王婧金,肖明虎,等.梗阻性肥厚型心肌病合并心房颤动的临床特征及危险因素[J].中华内科杂志,2017,56(3):184-187.
[14]中国医师协会心血管内科医师分会,《中华内科杂志》编辑委员会.心血管疾病一级预防中国专家共识[J].中华内科杂志,2010,49(2):174-185.
[15]SIONTIS KC,GESKE JB,ONG K,et al.Atrial fibrillation in hypertrophic cardiomyopathy:Prevalence,clinical correlations,and mortality in a large high-risk population[J].J Am Heart Assoc,2014,3(3):e001002.
[16]TAN AY,BLUMENFELD J,MICHAEEL A,et al.Autosomal dominant polycystic kidney disease caused by somatic and germline mosaicism[J].Clin Genet,2015,87(4):373-377.
[17]LAKDAWALA NK,FUNKE BH,BAXTER S,et al.Genetic testing for dilated cardiomyopathy in clinical practice[J].J Card Fail,2012,18(4):296-303.
[18]KLAASSEN S,PROBST S,OECHSLIN E,et al.Mutations in sarcomere protein genes in left ventricular noncompaction[J].Circulation,2008,117(22):2893-2901.
[2]MARON BJ,MARON MS.Hypertrophic cardiomyopathy[J].Lancet,2013,381(9862):242-255.
[3]ELLIOTT PM,ANASTASAKIS A,BORGER MA,et al.2014ESC Guidelines on Diagnosis and Management of Hypertrophic Cardiomyopathy:the Task Force for the Diagnosis and Management of Hypertrophic Cardiomyopathy of the European Society of Cardiology(ESC)[J].Eur Heart J,2014,35(39):2733-2779.
[4]KRAKER J,VISWANATHAN SK,KNOLL R,et al.Recent advances in the molecular genetics of familial hypertrophic cardiomyopathy in south Asian descendants[J].Front Physiol,2016,7:499.
[5]FU L,LUO S,CAI S,et al.Identification of LAMP2 mutations in early-onset danon disease with hypertrophic cardiomyopathy by targeted next-generation sequencing[J].Am J Cardiol,2016,118(6):888-894.
[6]MANGO R,LUCHETTI A,SANGIUOLO R,et al.Next generation sequencing and linkage analysis for the molecular diagnosis of a novel overlapping syndrome characterized by hypertrophic cardiomyopathy and typical electrical instability of Brugada syndrome[J].Circ J,2016,80(4):938-949.
[7]WEISSLER-SNIR A,CREAN A,RAKOWSKI H.The role of imaging in the diagnosis and management of hypertrophic cardiomyopathy[J].Expert Rev Cardiovasc Ther,2016,14(1):51-74.
[8]杨雪蓉,陈代文,余冰,等.PRDM16的研究进展[J].动物营养学报,2010,22(6):1477-481.
[9]PINHEIRO I,MARGUERON R,SHUKEIR N,et al.Prdm3and Prdm16 are H3K9me1 methyltransferases required for mammalian heterochromatin integrity[J].Cell,2012,150(5):948-960.
[10]BJORK BC,TURBE-DOAN A,PRYSAK M,et al.Prdm16is required for normal palatogenesis in mice[J].Hum Mol Genet,2010,19(5):774-789.
[11]ARNDT AK,SCHAFER S,DRENCKHAHN JD,et al.Fine mapping of the 1p36 deletion syndrome identifies mutation of PRDM16 as a cause of cardiomyopathy[J].Am J Hum Genet,2013,93(1):67-77.
[12]OLIVOTTO I,CECCHI F,CASEY SA,et al.Impact of atrial fibrillation on the clinical course of hypertrophic cardiomyopathy[J].Circulation,2001,104(21):2517-2524.
[13]伍熙崔,王婧金,肖明虎,等.梗阻性肥厚型心肌病合并心房颤动的临床特征及危险因素[J].中华内科杂志,2017,56(3):184-187.
[14]中国医师协会心血管内科医师分会,《中华内科杂志》编辑委员会.心血管疾病一级预防中国专家共识[J].中华内科杂志,2010,49(2):174-185.
[15]SIONTIS KC,GESKE JB,ONG K,et al.Atrial fibrillation in hypertrophic cardiomyopathy:Prevalence,clinical correlations,and mortality in a large high-risk population[J].J Am Heart Assoc,2014,3(3):e001002.
[16]TAN AY,BLUMENFELD J,MICHAEEL A,et al.Autosomal dominant polycystic kidney disease caused by somatic and germline mosaicism[J].Clin Genet,2015,87(4):373-377.
[17]LAKDAWALA NK,FUNKE BH,BAXTER S,et al.Genetic testing for dilated cardiomyopathy in clinical practice[J].J Card Fail,2012,18(4):296-303.
[18]KLAASSEN S,PROBST S,OECHSLIN E,et al.Mutations in sarcomere protein genes in left ventricular noncompaction[J].Circulation,2008,117(22):2893-2901.