Analysis of VWF Gene c.7332G＞A Nonsense Mutation Pedigree and Study of Molecular Pathogenesis
10.19746/j.cnki.issn1009-2137.2025.06.022
- VernacularTitle:VWF基因c.7332G＞A无义突变家系分析及致病机制研究
- Author:
Duan-Yang WANG
1
;
Lei WANG
1
;
Dong-Yan FU
1
;
Xiao-Mei LU
1
;
Li-Dong ZHAO
1
;
Jia-Wei ZHENG
1
;
Ya-Lin YU
1
;
Gang WANG
1
;
Lin-Hua YANG
1
Author Information
1. 山西医科大学第二临床医学院(第二医院)血液科,山西医科大学出凝血疾病及恶性血液病研究中心,山西太原 030001
- Publication Type:Journal Article
- Keywords:
VWF;
NGS;
nonsense mutation;
multimeric analysis;
pathogenesis
- From:
Journal of Experimental Hematology
2025;33(6):1701-1707
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To analyze the genetic characteristics of the VWF gene c.7332G＞A nonsense mutation and explore its molecular pathogenesis.Methods:Phenotypic diagnosis of the proband was performed using VWF:Ag,VWF:RCo,FⅧ:C and multimeric analysis.The probands were genotyped by NGS whole-exome sequencing,and the sequencing results were validated by sanger sequencing.The family members were genotyped by Sanger sequencing.The VWF gene c.7332G＞A nonsense mutant plasmid was constructed.After transfection,the function of VWF gene c.7332G＞A mutant plasmid was verified at cell level in vitro.The mRNA level was detected by qRT-PCR,and the expression level of protein was detected by Western blot,the function of multimerization was verified by the multimeric analysis.Results:VWF:Ag and VWF:RCo were all less than 3％in the proband,and the multimeric analysis showed multimer deficiency.The proband was diagnosed as type 3 VWD.The homozygous nonsense mutation of VWF gene c.7332G＞A was detected by gene sequencing.The VWF mRNA level of the mutant plasmid was decreased,and the VWF protein expression in the cell supernatant was decreased,the mutant protein was truncated and the function of VWF multimerization was impaired.Conclusion:A homozygous mutation in exon 43 of VWF gene,c.7332G＞A,was responsible for the probands type 3 VWD in the proband.The mutation caused a decrease in the relative level of VWF mRNA and protein,and impaired the function of VWF multimerization.
