云南不明原因猝死高发地区汉族人群易感基因变异筛查

Study of Yunnan sudden unexplained death in Han population and cardiac disease associated genetic variants

目的筛查云南不明原因猝死高发地区汉族人群的心脏疾病易感基因变异。方法采集96例云南不明原因猝死高发地区汉族村民(病区组)和183例邻村健康村民(对照组)的外周静脉血,提取基因组DNA,使用基质辅助激光解析电离时间飞行质谱技术(MALDI-TOF-MS)对19个心脏疾病相关基因的35个单核苷酸变异位点进行基因分型。采用SPSS17.0软件分析数据,并通过蛋白质功能预测软件PolyPhen-2、SIFT和MutationTaster对具有统计学差异的变异位点进行致病性预测。使用十二导联心电图机对所有个体进行心电图检测。结果 AKAP9(rs146797353,c.3827G>A,p.R1276Q)、AKAP9(rs73407505,c.4199T>C,p.M1400T)和RNF207(rs201773865,c.1616G>A,p.R539H)的等位基因频率及基因型频率在病区组与对照组之间具有统计学差异,且AKAP9(rs146797353)、AKAP9(rs73407505)在病区组中的等位基因频率及基因型频率均高于对照组。蛋白功能预测软件SIFT对R1276Q预测结果为影响蛋白质功能,SIFT和MutationTaster对R539H预测结果为可致病性。病区组中检出ST-T改变、T波异常、传导阻滞等异常心电图改变且频率高于对照组。结论 YNSUD高发地区汉族人群的心脏电生理异常可能是其发病的易感因素。YNSUD高发地区汉族人群中尚未发现明确的可致病性心脏疾病基因的可致病性变异,YNSUD在云南汉族人群高发的原因有待进一步研究。

Objective To explore the association of cardiac associated genetic variants and the high incidence of Yunnan sudden unexplained death(YNSUD) in Han population. Methods The genomic DNA was extracted from peripheral blood samples of 96 villagers from YNSUD aggregative villages(AV) and 183 villagers from neighboring control villages(CV). The genotyping was performed in 35 single nucleotide mutations of 19 cardiac disease associated genes, using MatrixAssisted Laser Desorption Ionization Time of Flight Mass Spectrometry(MALDI-TOF MS). The Statistical Program for Social Science(SPSS version 17.0, Chicago, IL) was used to carry out statistical analysis, and the pathogenicity of variants with statistical difference were predicted by silico prediction tool(PolyPhen-2、SIFT and MutationTaster). All participants were given ECG examination using a 12-lead machine at rest with standard lead positions. Results Statistical differences were observed in AKAP9(rs146797353, c.3827 G>A, p.R1276 Q) and AKAP9(rs73407505, c.4199 T>C, p.M1400 T) and RNF207(rs201773865, c.1616 G>A, p.R539 H) between two groups, and both allele frequency and genotype frequency of AKAP9(rs146797353, c.3827 G>A, p.R1276 Q) and AKAP9(rs73407505, c.4199 T>C, p.M1400 T) in AV were higher than those in CV. As protein function, R1276 Q was predicted to be affected protein function by SIFT, and R539 H was predicted to be a disease causing mutation by SIFT and Mutationtaster. ST-T alteration, T wave changes and heart-blocking were identified in AV and the frequency were higher than that in CV. Conclusion This study is the first genetic analysis of cardiac associated genes in Han population with high incidence of YNSUD and has not found definite pathogenic variants, but abnormal cardiac electrophysiology may be a susceptibility factor, which suggests that the cause of high incidence of YNSUD in Han population needs further study.