Objective: To analysis clinical phenotype and potential genetic cause of a family affected with hereditary coagulation factor Ⅻ deficiency.
Methods: The prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), D-Dimer (D-D), coagulation factor Ⅻ activity (FⅫ:C) and coagulation factor Ⅻ antigen (FⅫ:Ag) were determined for phenotype diagnosis of the proband and his family members(3 generations and 5 people). Targeted capture and whole exome sequencing were performed in peripheral blood sample of the proband. Possible disease-causing mutations of F12 gene were obtained and further confirmed by Sanger sequencing. The corresponding mutation sites of the family members were analyzed afterwards. The online bioinformatics software AutoPVS1 and Mutation Taster was used to predict the effects of mutation sites on protein function.
Results: The APTT of the proband was significantly prolonged, reaching 180.9s. FⅫ:C and FⅫ:Ag of the proband was significantly reduced to 0.8% and 4.17%, respectively. The results of whole exome sequencing displayed that there were compound heterozygous mutations in F12 gene of the proband, including the c.1261G＞T heterozygous nonsense mutation in exon 11 (causing p.Glu421*) and the c.251dupG heterozygous frameshift mutation in exon 4 (causing p.Trp85Metfs*53). Both mutations are loss of function mutations with very strong pathogenicity, leading to premature termination of the protein. AutoPVS1 and Mutation Taster software predicted both mutations as pathogenic mutations. The results of Sanger sequencing revealed that c.1261G＞T heterozygous mutation of the proband was inherited from his mother, for which his brother and his daughter were c.1261G＞T heterozygous carriers. Genotype-phenotype cosegregation was observed in this family.
Conclusion: The c.1261G＞T heterozygous nonsense mutation in exon 11 and the c.251dupG heterozygous frameshift mutation in exon 4 of the F12 gene probably account for coagulation factor Ⅻ deficiency in this family. This study reports two novel pathogenic F12 mutations for the first time worldwide.
题目: 复合杂合突变导致的遗传性凝血因子Ⅻ缺陷症家系分析.
目的: 对1个遗传性凝血因子Ⅻ（FⅫ）缺陷症家系进行临床表型及基因突变分析，探讨其分子致病机制.
方法: 检测先证者及其家系成员（共3代5人）血浆凝血酶原时间（PT）、活化部分凝血活酶时间（APTT）、纤维蛋白原含量（FIB）、D-D二聚体（D-D）、凝血因子Ⅻ促凝活性（FⅫ:C）和凝血因子Ⅻ抗原（FⅫ:Ag）等凝血指标。采用高通量测序方法分析先证者F12基因所有的外显子编码区及外显子-内含子交界处的基因突变情况，对检出的可疑致病突变进行Sanger测序验证，并对家系成员的相应突变位点进行检测。采用AutoPVS1和Mutation Taster在线生物信息学软件预测突变位点对蛋白功能的影响.
结果: 先证者APTT结果为180.9 s，明显延长；FⅫ:C和FⅫ:Ag分别降低至0.8%和4.17%。全外显子组测序分析发现先证者F12基因存在复合杂合突变，即第11号外显子c.1261G＞T杂合无义突变（p.Glu421*）和第4号外显子c.251dupG杂合移码突变（p.Trp85Metfs*53），均为功能缺失型突变，属于极强致病性级别。Sanger测序家系验证结果显示，先证者c.1261G＞T杂合突变遗传自其母亲，其哥哥和女儿均为c.1261G＞T杂合突变携带者。在此家系中基因检测结果与血液学检查结果相符，即基因型和表型共分离.
结论: F12基因第11号外显子c.1261G＞T杂合无义突变和第4号外显子c.251dupG杂合移码突变是本例家系遗传性凝血因子Ⅻ缺陷症的分子发病机制，这两个突变均为国际上首次报道的新突变.
Keywords: F12 gene; coagulation factor Ⅻ deficiency; novel mutation.