诊断学理论与实践2025，Vol.24Issue(4)：431-440,10.DOI:10.16150/j.1671-2870.2025.04.010
三种ADAMTS13去整合素样结构域突变致蛋白功能缺陷及其与血栓关联的研究
Three disintegrin-like domain mutations of ADAMTS13:functional deficiency and association with thrombosis
摘要
Abstract
Objective To analyze the genotypic and phenotypic characteristics of four thrombotic patients carrying heterozygous mutations in the disintegrin-like domain of ADAMTS13(a disintegrin and metalloproteinase with thrombospon-din type 1 motifs 13),and to explore the association between ADAMTS13 deficiency and thrombosis.Methods Thrombophilia-related gene mutations were screened in thrombosis patients using a thrombophilia gene panel,identifying four thrombophilia pedigrees carrying mutations in the ADAMTS13 disintegrin-like domain.Coagulation function was assessed using clotting tests.Antigen and activity of von Willebrand factor(vWF)were measured using immunoturbidime-try,and its collagen-binding capacity was measured using enzyme-linked immunosorbent assay(ELISA).The distribution of vWF multimers was analyzed by SDS-agarose gel electrophoresis with gray value semi-quantitative analysis.ADAMTS13 activity in plasma was measured by fluorescence resonance energy transfer assay,and its antigen levels were quantified via ELISA.Comparative analysis of three-dimensional structures between wild-type and mutant ADAMTS13 proteins was per-formed using PyMOL software.Results The probands from four pedigrees experienced thrombotic events of varying seve-rity,including cerebral venous sinus thrombosis,pulmonary embolism,and lower extremity deep vein thrombosis.Muta-tions in the ADAMTS13 disintegrin-like domain accounted for approximately 4/87 of ADAMTS13 heterozygous mutations.Genetic analysis showed that all four probands carried heterozygous mutations in the ADAMTS13 disintegrin-like domain(p.Pro301Ala,p.Pro301Arg,p.Arg349Cys),with p.Pro301Ala and p.Pro301Arg being newly reported.Coagulation tests demon-strated significantly reduced ADAMTS13 activity and antigen levels(Act:57.42％-72.88％,Ag:66.94％-78.34％),and elevated vWF activity and antigen levels(Act:158.2％-213.7％,Ag:167.2％-216.6％).Electrophoretic analysis of vWF multimers showed a significantly increased proportion of high-molecular-weight multimers(HMWMs)in patient plasma(Gray value:166.6-218.9 vs 117.4),indicating markedly elevated HMWMs compared to normal controls.Structural analysis further suggested that the mutation sites were located in critical regions of the ADAMTS13 disintegrin-like domain,potentially disrupting protein stability and its binding capacity with vWF.Conclusion This study first reports two novel missense muta-tions—Pro301Ala and Pro301Arg—located in the disintegrin-like domain of the ADAMTS13 protein,and further verifies the pathogenic characteristics of the known Arg349Cys mutation.The results demonstrate that these mutations result in reduced ADAMTS13 protein expression levels and significantly decreased enzymatic activity,as evidenced by impaired cleavage capacity of high-molecular-weight vWF multimers.Functional tests confirm abnormally increased vWF multimers in patients,disrupting the dynamic balance of the ADAMTS13-vWF axis and consequently increasing thrombotic risk.关键词
血管性血友病因子裂解酶13/血管性血友病因子/去整合素样结构域/基因突变/血栓形成Key words
ADAMTS13/von Willebrand factor/Disintegrin-like domain/Gene mutation/Thrombosis分类
医药卫生引用本文复制引用
林莉亚,吴希,毛胤祺,陈光明,武文漫,戴菁,王学锋,丁秋兰..三种ADAMTS13去整合素样结构域突变致蛋白功能缺陷及其与血栓关联的研究[J].诊断学理论与实践,2025,24(4):431-440,10.基金项目
国家自然科学基金面上项目(82170128) （82170128）
国家重点研发计划项目(2023YFC2507800) （2023YFC2507800）
