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A point mutation in GPI-attachment signal peptide accelerates the development of prion disease

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Abstract

A missense variant from methionine to arginine at codon 232 (M232R) of the prion protein gene accounts for ~ 15% of Japanese patients with genetic prion diseases. However, pathogenic roles of the M232R substitution for the induction of prion disease have remained elusive because family history is usually absent in patients with M232R. In addition, the clinicopathologic phenotypes of patients with M232R are indistinguishable from those of sporadic Creutzfeldt-Jakob disease patients. Furthermore, the M232R substitution is located in the glycosylphosphatidylinositol (GPI)-attachment signal peptide that is cleaved off during the maturation of prion proteins. Therefore, there has been an argument that the M232R substitution might be an uncommon polymorphism rather than a pathogenic mutation. To unveil the role of the M232R substitution in the GPI-attachment signal peptide of prion protein in the pathogenesis of prion disease, here we generated a mouse model expressing human prion proteins with M232R and investigated the susceptibility to prion disease. The M232R substitution accelerates the development of prion disease in a prion strain-dependent manner, without affecting prion strain-specific histopathologic and biochemical features. The M232R substitution did not alter the attachment of GPI nor GPI-attachment site. Instead, the substitution altered endoplasmic reticulum translocation pathway of prion proteins by reducing the hydrophobicity of the GPI-attachment signal peptide, resulting in the reduction of N-linked glycosylation and GPI glycosylation of prion proteins. To the best of our knowledge, this is the first time to show a direct relationship between a point mutation in the GPI-attachment signal peptide and the development of disease.

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Data availability

The data that support the findings are available from the corresponding author upon request. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD037584 and 10.6019/PXD037584.

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Acknowledgements

We thank members of the Creutzfeldt-Jakob Disease Surveillance Committee in Japan, Creutzfeldt-Jakob disease specialists in the prefectures, and Creutzfeldt-Jakob disease patients and families for providing important clinical information. We thank Hiroko Kudo, Miyuki Yamamoto, Ayumi Yamazaki, and Akinori Ninomiya for their excellent technical assistance.

Funding

This work was supported by JSPS KAKENHI Grant Number 18K05963 (AK), The Kato Memorial Trust for Nambyo Research (AK), Takeda Science Foundation (AK), The Akiyama Life Science Foundation (AK), The Ichiro Kanehara Foundation (AK), The Suhara Memorial Foundation (AK), JST ACT-X Grant Number JPMJAX201B (TH), and JSPS KAKENHI Grant Number 21H02415 (TKin).

Author information

Author notes

  1. Atsushi Kobayashi

    Present address: Department of Biomedical Models, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4, Sakamoto, Nagasaki, 852-8523, Japan

  2. Shirou Mohri

    Present address: Research Center for Biomedical Models and Animal Welfare, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, 852-8523, Japan

  3. Atsushi Kobayashi and Tetsuya Hirata have contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Comparative Pathology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido, 060-0818, Japan

    Atsushi Kobayashi, Taishi Shimazaki, Yoshiko Munesue, Keisuke Aoshima & Takashi Kimura

  2. Institute for Glyco-core Research (iGCORE), Gifu University, 501-1193, Gifu, Japan

    Tetsuya Hirata

  3. Laboratory of Histology and Cytology, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, Hokkaido, 060-8638, Japan

    Junko Nio-Kobayashi

  4. Biomedical Animal Research Laboratory, Institute for Genetic Medicine, Hokkaido University, Sapporo, Hokkaido, 060-0615, Japan

    Rie Hasebe

  5. Department of Neurological Science, Tohoku University Graduate School of Medicine, Sendai, Miyagi, 980-8575, Japan

    Atsuko Takeuchi, Shirou Mohri & Tetsuyuki Kitamoto

  6. National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, 305-0856, Japan

    Yuichi Matsuura

  7. Division of Morphological Sciences, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, 890-8544, Japan

    Satoshi Kusumi

  8. Department of Microscopic Anatomy and Cell Biology, Asahikawa Medical University, Asahikawa, Hokkaido, 078-8510, Japan

    Daisuke Koga

  9. Department of Neuropathology, Institute for Medical Science of Aging, Aichi Medical University, Nagakute, Aichi, 480-1195, Japan

    Yasushi Iwasaki

  10. Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, 565-0871, Japan

    Taroh Kinoshita

  11. WPI Immunology Frontier Research Center, Osaka University, Suita, Osaka, 565-0871, Japan

    Taroh Kinoshita

  12. Center for Infectious Disease Education and Research, Osaka University, Suita, Osaka, 565-0871, Japan

    Taroh Kinoshita

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  1. Atsushi Kobayashi
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Contributions

AK, TH: conceptualization; TKin, SM, TKit: methodology; AK, TH, TS, YMu, JNK, RH, AT, YMa, SK, DK, YI: investigation; AK, TH, TS, JNK, RH, SK, DK: visualization; AK, TH, TKin: funding acquisition; AK: project administration; KA, TKim, TKin, SM, TKit: Supervision; AK, TH, TS: writing—original draft; KA, TKim, TKin, SM, TKit: writing—review & editing.

Corresponding author

Correspondence to Atsushi Kobayashi.

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The authors report no competing interests.

Ethical approval

This study was approved by the Institutional Ethics Committee of Hokkaido University Faculty of Veterinary Medicine (VET27-1, VET1-5). All experiments using human materials were in compliance with the Helsinki Declaration. Animal experiments were performed in strict accordance with the Regulations for Animal Experiments and Related Activities at Hokkaido University and Fundamental Guidelines for Proper Conduct of Animal Experiment and Related Activities in Academic Research Institutions by the Ministry of Education, Culture, Sports, Science and Technology in Japan, Notice No. 71. Protocols for animal experiments were approved by the Institutional Animal Care and Use Committees of Hokkaido University (14-0170, 19-0025).

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Kobayashi, A., Hirata, T., Shimazaki, T. et al. A point mutation in GPI-attachment signal peptide accelerates the development of prion disease. Acta Neuropathol 145, 637–650 (2023). https://doi.org/10.1007/s00401-023-02553-5

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