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The fibrous form of intracellular inclusion bodies in recombinant variant fibrinogen-producing cells is specific to the hepatic fibrinogen storage disease-inducible variant fibrinogen

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Abstract

Fibrinogen storage disease (FSD) is a rare disorder that is characterized by the accumulation of fibrinogen in hepatocytes and induces liver injury. Six mutations in the γC domain (γG284R, γT314P, γD316N, the deletion of γG346-Q350, γG366S, and γR375W) have been identified for FSD. Our group previously established γ375W fibrinogen-producing Chinese hamster ovary (CHO) cells and observed aberrant large granular and fibrous forms of intracellular inclusion bodies. The aim of this study was to investigate whether fibrous intracellular inclusion bodies are specific to FSD-inducible variant fibrinogen. Thirteen expression vectors encoding the variant γ-chain were stably or transiently transfected into CHO cells expressing normal fibrinogen Aα- and Bβ-chains or HuH-7 cells, which were then immunofluorescently stained. Six CHO and HuH-7 cell lines that transiently produced FSD-inducible variant fibrinogen presented the fibrous (3.2–22.7 and 2.1–24.5%, respectively) and large granular (5.4–25.5 and 7.7–23.9%) forms of intracellular inclusion bodies. Seven CHO and HuH-7 cell lines that transiently produced FSD-non-inducible variant fibrinogen only exhibit the large granular form. These results demonstrate that transiently transfected variant fibrinogen-producing CHO cells and inclusion bodies of the fibrous form may be useful in non-invasive screening for FSD risk factors for FSD before its onset.

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Acknowledgements

This work was supported by JSPS KAKENHI Grant Number 26460672 (N. Okumura).

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Authors and Affiliations

  1. Department of Laboratory Medicine, Graduate School of Medicine, Shinshu University, Matsumoto, Japan

    Shinpei Arai & Takayuki Honda

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

    Shinpei Arai, Naoko Ogiwara, Saki Mukai, Yuka Takezawa & Mitsutoshi Sugano

  3. Department of Health and Medical Sciences, Graduate School of Medicine, Shinshu University, Matsumoto, Japan

    Nobuo Okumura

  4. Laboratory of Clinical Chemistry and Immunology, Department of Biomedical Laboratory Sciences, School of Health Sciences, Shinshu University, 3-1-1 Asahi, Matsumoto, 390-8621, Japan

    Nobuo Okumura

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  1. Shinpei Arai
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  2. Naoko Ogiwara
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  3. Saki Mukai
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  4. Yuka Takezawa
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  5. Mitsutoshi Sugano
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  6. Takayuki Honda
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  7. Nobuo Okumura
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Correspondence to Nobuo Okumura.

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Arai, S., Ogiwara, N., Mukai, S. et al. The fibrous form of intracellular inclusion bodies in recombinant variant fibrinogen-producing cells is specific to the hepatic fibrinogen storage disease-inducible variant fibrinogen. Int J Hematol 105, 758–768 (2017). https://doi.org/10.1007/s12185-017-2185-5

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  • DOI: https://doi.org/10.1007/s12185-017-2185-5

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