A series of 10 Polish patients with thromboembolic events and antithrombin deficiency

Wójcik, Magdalena; Morena‐Barrio, María Eugenia de la; Michalik, Justyna; Wypasek, Ewa; Kopytek, M; Corral, Javier; Undas, Anetta

doi:10.1097/mbc.0000000000000816

Cited by 5 publications

(1 citation statement)

References 32 publications

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“…Type I: Classical deficiency, characterized by impaired AT synthesis and a parallel decrease in plasma AT antigen and activity in patients. Its clinical manifestations are severe and usually associated with severe mutations in SER-PINC1, which lead to a significant decrease in the level of AT in plasma owing to mutations that destabilize mRNA, misfolded proteins, and intracellular retention, or lead to abnormal degradation [21][22][23]. Type II mainly affects the domain function bound to thrombin or heparin, with normal plasma AT antigen content, but its activity is weakened [24], its clinical manifestations are mild, and its incidence is relatively high [25].…”

Section: Discussionmentioning

confidence: 99%

Identification and characterization of two SERPINC1 mutations causing congenital antithrombin deficiency

Wang

Ruan

et al. 2023

Thrombosis J

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Background Antithrombin (AT) is the main physiological anticoagulant involved in hemostasis. Hereditary AT deficiency is a rare autosomal dominant thrombotic disease mainly caused by mutations in SERPINC1, which was usually manifested as venous thrombosis and pulmonary embolism. In this study, we analyzed the clinical characteristics and screened for mutant genes in two pedigrees with hereditary AT deficiency, and the functional effects of the pathogenic mutations were evaluated. Methods Candidate gene variants were analyzed by next-generation sequencing to screen pathogenic mutations in probands, followed by segregation analysis in families by Sanger sequencing. Mutant and wild-type plasmids were constructed and transfected into HEK293T cells to observe protein expression and cellular localization of SERPINC1. The structure and function of the mutations were analyzed by bioinformatic analyses. Results The proband of pedigree A with AT deficiency carried a heterozygous frameshift mutation c.1377delC (p.Asn460Thrfs*20) in SERPINC1 (NM000488.3), a 1377C base deletion in exon 7 resulting in a backward shift of the open reading frame, with termination after translation of 20 residues, and a different residue sequence translated after the frameshift. Bioinformatics analysis suggests that the missing amino acid sequence caused by the frameshift mutation might disrupt the disulfide bond between Cys279 and Cys462 and affect the structural function of the protein. This newly discovered variant is not currently included in the ClinVar and HGMD databases. p.Arg229* resulted in a premature stop codon in exon 4, and bioinformatics analysis suggests that the truncated protein structure lost its domain of interaction with factor IX (Ala414 site) after the deletion of nonsense mutations. However, considering the AT truncation protein resulting from the p.Arg229* variant loss a great proportion of the molecule, we speculate the variant may affect two functional domains HBS and RCL and lack of the corresponding function. The thrombophilia and decreased-AT-activity phenotypes of the two pedigrees were separated from their genetic variants. After lentiviral plasmid transfection into HEK293T cells, the expression level of AT protein decreased in the constructed c.1377delC mutant cells compared to that in the wild-type, which was not only reduced in c.685C > T mutant cells but also showed a significant band at 35 kDa, suggesting a truncated protein. Immunofluorescence localization showed no significant differences in protein localization before and after the mutation. Conclusions The p.Asn460Thrfs*20 and p.Arg229* variants of SERPINC1 were responsible for the two hereditary AT deficiency pedigrees, which led to AT deficiency by different mechanisms. The p.Asn460Thrfs*20 variant is reported for the first time.

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Section: Discussionmentioning

confidence: 99%

Identification and characterization of two SERPINC1 mutations causing congenital antithrombin deficiency

Wang

Ruan

et al. 2023

Thrombosis J

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Analysis of phenotype and gene mutation in three pedigrees with inherited antithrombin deficiency

Jiang

Liu

et al. 2022

Clinical Laboratory Analysis

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Background Inherited AT deficiency is an autosomal‐dominant thrombophilic disorder usually caused by various SERPINC1 defects associated with a high risk of recurrent venous thromboembolism. In this article, the phenotype, gene mutation, and molecular pathogenic mechanisms were determined in three pedigrees with inherited AT deficiency. Methods Coagulation indices were examined on STAGO STA‐R‐MAX analyzer. The AT:Ag was analyzed by ELISA. All exons and flanking sequences of SERPINC1 were amplified by PCR. AT wild type and three mutant expression plasmids were constructed and then transfected into HEK293FT cells. The expression level of AT protein was analyzed by ELISA and Western blot. Results The AT:A and AT:Ag of probands 1 and 3 were decreased to 49% and 52 mg/dL, 38% and 44 mg/dL, respectively. The AT:A of proband 2 was decreased to 32%. The SERPINC1 gene analysis indicated that there was a p.Ile421Thr in proband 1, a p.Leu417Gln in proband 2, and a p.Met252Thr in proband 3, respectively. The AT mRNA expression level of the three mutants was not significantly different from AT‐WT by qRT‐PCR. The results of ELISA and Western blot tests showed that the AT‐M252T and AT‐I421T mutants had a higher AT expression than the AT wild type (AT‐WT), and the AT protein expression of AT‐L417Q mutants had no significant difference compared with AT‐WT in the cell lysate. The AT expression levels of AT‐M252T and AT‐I421T mutants were lower than that of AT‐WT, and there was no significant difference between AT‐L417Q mutant and AT‐WT in the supernatant. Conclusion The p.I421T and p.M252T mutations affected the secretion of AT protein leading to type I AT deficiency of probands 1 and 3. The p.Leu417Gln mutation was responsible for the impaired or ineffective activity AT protein in proband 2 and caused type II AT deficiency.

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Generation of a human iPSC line ZZUNEUi014-A from a patient with antithrombin deficiency caused by mutation in SERPINC1 gene

Hong

Xing

et al. 2022

Stem Cell Research

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A series of 10 Polish patients with thromboembolic events and antithrombin deficiency

Cited by 5 publications

References 32 publications

Identification and characterization of two SERPINC1 mutations causing congenital antithrombin deficiency

Identification and characterization of two SERPINC1 mutations causing congenital antithrombin deficiency

Analysis of phenotype and gene mutation in three pedigrees with inherited antithrombin deficiency

Generation of a human iPSC line ZZUNEUi014-A from a patient with antithrombin deficiency caused by mutation in SERPINC1 gene

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