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Novel variant fibrinogen γp.C352R produced hypodysfibrinogenemia leading to a bleeding episode and failure of infertility treatment

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Abstract

Introduction

We 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 methods

Patient 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.

Results

Purified 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.

Conclusion

We 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.

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Acknowledgements

We gratefully acknowledge Dr. Takashi Ushiki (Department of Transfusion Medicine, Cell Therapy and Regenerative Medicine, Niigata University Medical and Dental Hospital, Japan) and Dr. Yoshinobu Seki (Department of Hematology, Uonuma Institute of Community Medicine, Niigata University Medical and Dental Hospital, Japan) for the patient referral of Niigata II. This work was supported by JSPS KAKENHI Grant Number JP20K07799 (Nobuo Okumura).

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Authors and Affiliations

  1. Department of Clinical Laboratory Investigation, Graduate School of Medicine, Shinshu University, 3-1-1 Asahi, Matsumoto, 390-8621, Japan

    Masahiro Yoda, Takahiro Kaido, Chiaki Taira, Yumiko Higuchi, Shinpei Arai & Nobuo Okumura

  2. Department of Medical Sciences, Graduate School of Medicine, Science and Technology, Shinshu University, Matsumoto, Japan

    Tomu Kamijo

  3. Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Japan

    Tomu Kamijo

  4. Department of Clinical Laboratory Sciences, School of Health Sciences, Shinshu University, Matsumoto, Japan

    Nobuo Okumura

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  1. Masahiro Yoda
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  2. Takahiro Kaido
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  3. Tomu Kamijo
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  4. Chiaki Taira
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  5. Yumiko Higuchi
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  6. Shinpei Arai
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  7. Nobuo Okumura
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Contributions

Masahiro Yoda, Takahiro Kaido, Tomu Kamijo performed the experiments. Masahiro Yoda wrote the paper. Chiaki Taira, Yumiko Higuchi, Shinpei Arai and Nobuo Okumura designed the research and discussed the data. Shinpei Arai and Nobuo Okumura reviewed the paper.

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Correspondence to Shinpei Arai.

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Yoda, M., Kaido, T., Kamijo, T. et al. Novel variant fibrinogen γp.C352R produced hypodysfibrinogenemia leading to a bleeding episode and failure of infertility treatment. Int J Hematol 114, 325–333 (2021). https://doi.org/10.1007/s12185-021-03174-y

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  • DOI: https://doi.org/10.1007/s12185-021-03174-y

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