Abstract
Hypodysfibrinogenemia is the least frequently reported congenital fibrinogen disorder, characterized by both quantity and quality defects of fibrinogen. In this study, we investigated the molecular basis of hypodysfibrinogenemia in a Chinese family. Functional fibrinogen was measured by Clauss method, and the antigenic fibrinogen was measured by immunoturbidimetry assay. All the exons and exon–intron boundaries of fibrinogen genes (FGA, FGB and FGG) were analysed by direct DNA sequencing. To further evaluate its molecular and functional characterizations, fibrinogen was purified from the plasma of propositus, then SDS-PAGE, fibrin polymerization, clot lysis, and electron microscopy scanning were all performed. The propositus showed a slight decrease of immunologic fibrinogen (1.52 g/L) but dramatically reduced functional fibrinogen (0.3 g/L). DNA sequencing revealed a novel heterozygous CCTTTGATG deletion in the exon 8 of FGG, leading to the deletion of Ala289, Phe290, and Asp291 in fibrinogen γ-chain. The polymerization of the fibrinogen from the propositus was markedly impaired, with prolonged lag period and decreased final turbidity. The fibrinogen clottability showed a reduced fraction of participating clot formation. While the clot lysis showed normal. Scanning electron microscopy revealed that the fibers of the propositus were thicker than normal, with larger pores and curlier meshworks. We conclude that γAla289_Asp291del is responsible for the hypodysfibrinogenemia in this case.
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Acknowledgements
We thank the members of this family for participation in the study. This study was supported by the National Natural Science Foundation of China (81501810; 81672087) and by Natural Science Foundation of Zhejiang Province (LQ15H200001).
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11239_2018_1678_MOESM2_ESM.tif
Supplementary material 2 Fig. S1 Analysis of urokinase-catalyzed plasmin digestion of fibrinogen. (A) The fibrinogen was incubated with plasminogen and urokinase in the presence of calcium. Samples were heated at 100 °C in the presence of 2% SDS and 10 mM DTT for 5 minutes to terminate the reaction after incubation for 2 h. All samples were run by SDS-PAGE and stained by Coomassie Brilliant Blue R-250. Lane 1: the supernatant of fibrinogen from patient incubated with plasminogen and urokinase. Lane 2: the fibrinogen from health control incubated with plasminogen and urokinase. Lane 3: Normal fibrinogen without incubation with plasminogen and urokinase as a normal control. Lane 4: the mixture of plasminogen and urokinase. (TIF 1303 KB)
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Zhu, L., Zhao, M., Wang, M. et al. Congenital hypodysfibrinogenemia associated with a novel deletion of three residues (γAla289_Asp291del) in fibrinogen. J Thromb Thrombolysis 46, 211–218 (2018). https://doi.org/10.1007/s11239-018-1678-2
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DOI: https://doi.org/10.1007/s11239-018-1678-2