Novel variant fibrinogen γ p.C352R produced hypodysfibrinogenemia leading to a bleeding episode and failure of infertility treatment
Novel variant fibrinogen γ p.C352R produced hypodysfibrinogenemia leading to a bleeding episode and failure of infertility treatment
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新型变异纤维蛋白原 γ p.C352R 产生低纤维蛋白原血症，导致出血事件和不孕症治疗失败
DOI：
10.1007/s12185-021-03174-y
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Okumura N.
中科院分区：
文献类型：
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作者：
Yoda M;Kaido T;Kamijo T;Taira C;Higuchi Y;Arai S;Okumura N.
我们发现了一个新的杂合子变异纤维蛋白原，γ p.C352 R（新泻II; N-II），谁有出血发作和不孕症治疗失败，并怀疑有低异常纤维蛋白原血症的基础上低和不一致的纤维蛋白原水平（功能测定0.33 g/L，免疫测定0.91 g/L）。我们分析了这种罕见的先天性纤维蛋白原disorder.Materials和MethodsPatient血浆纤维蛋白原的表型的机制进行纯化和蛋白质表征和凝血酶催化的纤维蛋白聚合。建立了重组产纤维蛋白原的中国仓鼠卵巢（CHO）细胞，用ELISA和Westernblotting分析了变异纤维蛋白原的组装和分泌。结果纯化的N-II血浆纤维蛋白原在正常γ链以下有一条小的低分子量条带，纤维蛋白聚合略有减少。在已建立的CHO细胞系中，有一定比例的p.C352R纤维蛋白原被分泌到培养液中，但p.C352R的γ-链被合成，变异纤维蛋白原在细胞内被组装。结论我们证明，纤维蛋白原N-II，γp.C352R与CHO细胞中变异纤维蛋白原的分泌显著减少有关，纯化血浆纤维蛋白原的纤维蛋白聚合仅受到轻微影响，并且纤维蛋白原N-II在血浆中产生低异常纤维蛋白原血症。
IntroductionWe identified a patient with a novel heterozygous variant fibrinogen, γp.C352R (Niigata II; N-II), who had a bleeding episode and failed infertility treatment and was suspected to have hypodysfibrinogenemia based on low and discordant fibrinogen levels (functional assay 0.33 g/L, immunological assay 0.91 g/L). We analyzed the mechanism of this rare phenotype of a congenital fibrinogen disorder.Materials and methodsPatient plasma fibrinogen was purified and protein characterization and thrombin-catalyzed fibrin polymerization performed. Recombinant fibrinogen-producing Chinese hamster ovary (CHO) cells were established and the assembly and secretion of variant fibrinogen analyzed by ELISA and western blotting.ResultsPurified N-II plasma fibrinogen had a small lower molecular weight band below the normal γ-chain and slightly reduced fibrin polymerization. A limited proportion of p.C352R fibrinogen was secreted into the culture medium of established CHO cell lines, but the γ-chain of p.C352R was synthesized and variant fibrinogen was assembled inside the cells.ConclusionWe demonstrated that fibrinogen N-II, γp.C352R was associated with markedly reduced secretion of variant fibrinogen from CHO cells, that fibrin polymerization of purified plasma fibrinogen was only slightly affected, and that fibrinogen N-II produces hypodysfibrinogenemia in plasma.