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Disease-Associated Q159X Mutant Prion Protein Is Sufficient to Cause Fatal Degenerative Disease in Mice

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Abstract

PRNP Q160X is one of the five dominantly inheritable nonsense mutations causing familial prion diseases. Till now, it remains unclear how this type of nonsense mutations causes familial prion diseases with unique clinical and pathological characteristics. Human prion protein (PrP) Q160X mutation is equivalent to Q159X in mouse PrP, which produces the mutant fragment PrP1-158. Through intracerebroventricular injection of recombinant adeno-associated virus in newborn mice, we successfully overexpressed mouse PrP1-158-FLAG in the central nervous system. Interestingly, high level PrP1-158-FLAG expression in the brain caused death in these mice with an average survival time of 60 ± 9.1 days. Toxicity correlated with levels of PrP1-158-FLAG but was independent of endogenous PrP. Histopathological analyses showed microgliosis and astrogliosis in mouse brains expressing PrP1-158-FLAG and most of PrP1-158-FLAG staining appeared intracellular. Biochemical characterization revealed that the majority of PrP1-158-FLAG were insoluble and a significant part of PrP1-158-FLAG appeared to contain an un-cleaved signal peptide that may contribute to its cytoplasmic localization. Importantly, an ~10-kDa proteinase K-resistant PrP fragment was detected, which was the same as those observed in patients suffering from this type of prion diseases. To our knowledge, this is the first animal study of familial prion disease caused by Q159X that recapitulates key features of human disease. It will be a valuable tool for investigating the pathogenic mechanisms underlying familial prion diseases caused by nonsense mutations.

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Acknowledgements

We thank Dr Yong-Sun Kim at Ilsong Institute of Life Science, Hallym University, Korea, for the generous gift of 3F10 anti-PrP antibody and Dr Chaoyang Li at Guangzhou Medical University for the generous gift of 8B4 anti-PrP antibody.

Funding

This work was supported by the internal funding from the Chinese Institute for Brain Research, Beijing.

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  1. Yan Zhang and Runchuan Yan contributed equally to this work.

Authors and Affiliations

  1. School of Basic Medical Sciences, Capital Medical University, Beijing, 100069, China

    Yan Zhang & Jiyan Ma

  2. College of Biological Sciences, China Agricultural University, Beijing, 100193, China

    Runchuan Yan

  3. Chinese Institute for Brain Research, Beijing, 102206, China

    Yan Zhang, Runchuan Yan, Xiangyi Zhang & Jiyan Ma

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Contributions

Conceptualization, J. M.; methodology, Y. Z., R. Y., and J. M.; experimentation, Y. Z., R. Y., and X. Z.; data analysis, Y. Z., R. Y., and J. M.; writing—original draft preparation, J. M., Y. Z.; writing—review and editing, X. Z., Y. Z., R. Y., and J. M.; funding acquisition, J. M.

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Correspondence to Jiyan Ma.

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12035_2024_4224_MOESM1_ESM.docx

Supplementary file1 Supplementary Fig. 1 The expression pattern of PrP1-158-FLAG detected by anti-FLAG antibody differs from that detected by anti-PrP antibodies. A. A schematic diagram illustrating the antigenic epitopes of PrP1-158-FLAG recognized by different PrP antibodies. B. WB analysis with different antibodies to detect the PrP1-158-FLAG expression in the brains injected with rAAV (batch 2).C. WB analysis with 6D11 antibody and anti-FLAG antibody to compare the PrP1-158-FLAG expression pattern in the brains injected with rAAV (batch 1).“WT” and “KO” represent mice with Prnp+/+ and Prnp-/- genotypes, respectively. Supplementary Fig. 2 Verification of the lack of PrP expression in Prnp-/- mice. A. A schematic diagram illustrating the antigenic epitopes of full length PrP recognized by different PrP antibodies. B. WB analysis with different antibodies to detect the expression of PrP in Prnp-/- (KO) and Prnp+/+ (WT) mice. (DOCX 1098 KB)

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Zhang, Y., Yan, R., Zhang, X. et al. Disease-Associated Q159X Mutant Prion Protein Is Sufficient to Cause Fatal Degenerative Disease in Mice. Mol Neurobiol (2024). https://doi.org/10.1007/s12035-024-04224-2

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