一个遗传性凝血因子Ⅹ缺陷症家系的临床特征与基因突变分析  

Clinical characteristics and gene analysis of a pedigree with hereditary coagulation factorⅩdeficiency caused by compound heterozygous mutations Arg327Trp and Cys121Arg

作　　者：虞丹丹[1] 徐琦煜[1] 邹安庆[1] 王明山[1] 杨丽红[1] 蒋淑婷 金艳慧[1] YU Dandan;XU Qiyu;ZOU Anqing;WANG Mingshan;YANG Lihong;JIANG Shuting;JIN Yanhui(Department of Clinical Laboratory,the 1st Affiliated Hospital of Wenzhou Medical University,Wenzhou,Zhejiang 325015,China)

机构地区：[1]温州医科大学附属第一医院医学检验中心,浙江温州325015

出　　处：《中国优生与遗传杂志》2023年第2期340-344,共5页Chinese Journal of Birth Health & Heredity

基　　金：温州市科技局(Y20180295)。

摘　　要：目的探讨一个遗传性凝血因子Ⅹ(FⅩ)缺陷症家系的临床特征与基因突变关系。方法调查家系,采用Sanger测序法分析先证者F10基因8个外显子及其侧翼序列,并对家系成员相同突变位点进行检测。通过凝血酶生成试验评估家系成员FⅩ的促凝活性。ClustalX-2.1-win和在线生物信息学软件(PolyPhen-2、SIFT和Mutation Taster)分别用于分析突变位点氨基酸的保守性和致病性;Swiss-PdbViewer软件用于蛋白质空间结构和突变氨基酸相互作用分析。结果先证者FⅩ活性(FⅩ:C)和FⅩ抗原(FⅩ:Ag)分别降低至7%和14%(参考范围分别为:90%~114%和90%~110%);先证者平素无自发性出血现象,外伤后也无明显出血异常。基因测序发现先证者F10基因第4号外显子存在c.361T>C(p.Cys121Arg)杂合错义突变及第8号外显子存在c.979C>T(p.Arg327Trp)杂合错义突变;其母亲和女儿为p.Arg327Trp的杂合子,二姐和儿子为p.Cys121Arg的杂合子。先证者凝血酶生成潜力比值和峰高比值下降,而其延迟时间和达峰时间延长。保守性分析显示,p.Arg327和p.Cys121位点在同源物种间均高度保守。生物信息学软件预测两种突变均为致病突变。突变蛋白模型分析显示,p.Arg327Trp突变后,导致蛋白质空间结构和稳定性发生改变。结论该先证者F10基因p.Arg327Trp和p.Cys121Arg复合杂合突变与其FⅩ水平减低有关;该复合杂合突变与临床出血症状相关性不明显。Objective Phenotypic and genotypic analysis were performed in a pedigree with hereditary coagulation factorⅩ(FⅩ)deficiency to search for the pathogenic genes and investigate their molecular pathogenesis.Methods FⅩactivity(FⅩ:C)and FⅩantigen(FⅩ:Ag)of family members were detected to confirm phenotype diagnosis.Eight exons and flanking sequences of the F10 gene in the proband were analyzed by sanger sequencing,and the same variable sites were indentified in other members of this family.Thrombin generation test was used to evaluate the procoagulant activity of the family members.ClustalX-2.1-win and three online bioinformatics software were used to analyze the conservatism and patho genicity of mutant amino acids,respectively.Swiss-Pdb Viewer software was used for analysis of protein spatial structure and mutant amino acids interaction.Results FⅩ:C and FⅩ:Ag of the proband were reduced to 7%and 14%,respectively.FⅩ:C and FⅩ:Ag of her mother,second sister,son,and daughter were decreased to about 50%of the normal control.Genetic analy sis showed that the proband has heterozygous missense mutations c.361T>C(p.Cys121Arg)and c.979C>T(p.Arg327Trp)in exon 4 and 8 of the F10 gene,respectively.Her mother and daughter were heterozygous of p.Arg327Trp,and her son and second sister were heterozygous of p.Cys121Arg.The mutant protein in the propositus had obstacles in the thrombin generation with the prolonged ratio of lag time,time to peak and the reduced ratio of endogenous thrombin potential,peak height.Results of conservative analysis showed that p.Arg327 and p.Cys121 sites were deeply conservative in homologous species.Online bioinformatics software predicted that both mutations were damaged.Protein mutant model analysis revealed that the spatial structure and stability of p.Arg327Trp were changed after mutation.These changes might result in the change of protein spatial structure and stability.Conclusion The compound heterozygous mutations p.Arg327Trp and p.Cys121Arg of the F10gene may be related to the d

关 键 词：凝血因子Ⅹ缺陷症 基因突变 凝血酶生成试验 生物信息学 

分 类 号：R596.1[医药卫生—内科学]