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医学部 医学科 感染症学講座微生物学分野 |
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論文 【 表示 ／ 非表示 】
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Takatsuki H, Imamura M, Mori T, Atarashi R
Sci Rep 12 ( 1 ) 7923 2022年5月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Scientific Reports
Each prion strain has its own characteristics and the efficacy of anti-prion drugs varies. Screening of prion disease therapeutics is typically evaluated by measuring amounts of protease-resistant prion protein (PrP-res). However, it remains unclear whether such measurements correlate with seeding activity, which is evaluated by real-time quaking-induced conversion (RT-QuIC). In this study, the effects of anti-prion compounds pentosan polysulfate (PPS), Congo red, and alprenolol were measured in N2a58 cells infected with Fukuoka-1 (FK1) or 22L strain. The compounds abolished PrP-res and seeding activity, except for N2a58/FK1 treated with PPS. Interestingly, the seeding activity of N2a58/FK1, which was reduced in the presence of PPS, was not lost and remained at low levels. However, upon removal of PPS, both were gradually restored to their original levels. These results indicate that low-level persistent prion infection keeping measurable seeding activity is induced by PPS in a strain-dependent manner. Furthermore, for protein misfolding cyclic amplification (PMCA), the anti-prion effect of PPS decreased in FK1 compared to 22L, suggesting that the differences occur at the level of the direct conversion. Our findings demonstrate that the advantages of RT-QuIC and PMCA can be exploited for more accurate assessment of therapeutic drug screening, reflecting strain differences.
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Imamura M, Tabeta N, Iwamaru Y, Takatsuki H, Mori T, Atarashi R
Biochem Biophys Res Commun 613 67 - 72 2022年5月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Biochemical and Biophysical Research Communications
Prion diseases are transmissible and progressive neurodegenerative disorders characterized by abnormal prion protein (PrPSc) accumulation in the central nervous system. Generation of synthetic PrPSc in a cell-free conversion system and examination of its transmissibility to animals would facilitate testing of the protein-only hypothesis and the understanding of the molecular basis of sporadic prion diseases. In this study, we used recombinant prion protein from a baculovirus-insect cell expression system (Bac-rPrP) and insect cell-derived cofactors to determine whether Bac-rPrPSc is spontaneously produced in intermittent ultrasonic reactions. No spontaneous generation of Bac-rPrPSc was observed at 37 °C, but when the reaction temperature was increased to 45 °C, Bac-rPrPSc was generated in all trials. Some Bac-rPrPSc variants were transmissible to mice, but when the reaction was repeated for 40 rounds, the transmissibility was lost. Notably, a variety of Bac-rPrPSc variants, including non-transmissible ones, differing in resistance to proteinase K and cofactor dependence during amplification, was generated under the same experimental conditions, including the same sonication settings and cofactors. However, their characteristics also disappeared after 40 reaction rounds and the variety converged onto a single variant. These results indicate that various Bac-rPrPSc variants with different transmissibility to mice and structural properties are generated, which compete with each other and gradually converge onto a variant with a slightly faster amplification rate.
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Ethanolamine is a new anti‐prion compound
Uchiyama K., Hara H., Chida J., Pasiana A.D., Imamura M., Mori T., Takatsuki H., Atarashi R., Sakaguchi S.
International Journal of Molecular Sciences 22 ( 21 ) 2021年11月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：International Journal of Molecular Sciences
Prion diseases are a group of fatal neurodegenerative disorders caused by accumulation of proteinaceous infectious particles, or prions, which mainly consist of the abnormally folded, am-yloidogenic prion protein, designated PrPSc. PrPSc is produced through conformational conversion of the cellular isoform of prion protein, PrPC, in the brain. To date, no effective therapies for prion diseases have been developed. In this study, we incidentally noticed that mouse neuroblastoma N2a cells persistently infected with 22L scrapie prions, termed N2aC24L1‐3 cells, reduced PrPSc levels when cultured in advanced Dulbecco’s modified eagle medium (DMEM) but not in classic DMEM. PrPC levels remained unchanged in prion‐uninfected parent N2aC24 cells cultured in advanced DMEM. These results suggest that advanced DMEM may contain an anti‐prion compound(s). We then successfully identified ethanolamine in advanced DMEM has an anti‐prion activity. Ethanola-mine reduced PrPSc levels in N2aC24L1‐3 cells, but not PrPC levels in N2aC24 cells. Also, oral administration of ethanolamine through drinking water delayed prion disease in mice intracerebrally inoculated with RML scrapie prions. These results suggest that ethanolamine could be a new anti-prion compound.
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Ethanolamine is a new anti-prion compound. 査読あり 国際共著
Uchiyama K, Hara H, Chida J, Pasiana AD, Imamura M, Mori T, Takatsuki H, Atarashi R, Sakaguchi S
Int J Mol Sci 22 ( 21 ) 11742 2021年10月
記述言語：英語 掲載種別：研究論文（学術雑誌）
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Ubagai K, Fukuda S, Mori T, Takatsuki H, Taguchi Y, Kageyama S, Nishida N, Atarashi R
Biochem Biophys Res Commun 526 ( 4 ) 1049 - 1053 2020年6月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Biochemical and Biophysical Research Communications
Real-time quaking-induced conversion (RT-QUIC) assays using Escherichia coli-derived purified recombinant prion protein (rPrP) enable us to amplify a trace amount of the abnormal form of PrP (PrPSc) from specimens. This technique can be useful for the early diagnosis of both human and animal prion diseases and the assessment of prion contamination. In the present study, we demonstrated that there are strain-specific differences in the RT-QUIC reactions between an atypical form of bovine spongiform encephalopathy (BSE), L-BSE, and classical BSE (C-BSE). Whereas mouse rPrP (rMoPrP) was efficiently converted to amyloid fibrils in the presence of PrPSc seed derived from either L-BSE or C-BSE, hamster rPrP (rHaPrP) was converted only in L-BSE, not C-BSE. These characteristics were preserved in the second round reaction, but gradually weakened in the subsequent rounds and were completely lost by the fifth round, most likely due to the selective growth advantage of nonspecific rPrP amyloid fibrils in the RT-QUIC. Our findings further enhance the discrimination of prion strains using RT-QUIC, and further our understanding of the molecular basis of prion strains.
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バキュロウイルス-昆虫細胞発現組換えPrP を用いた試験管内変換系の構築に基づくプリオン変換・生成機構の解明. 招待あり
今村守一，森剛志，高月英恵，岩丸祥史，新竜一郎
YAKUGAKU ZASSHI 139 ( 7 ) 989 - 992 2019年7月
記述言語：日本語 掲載種別：記事・総説・解説・論説等（学術雑誌） 出版者・発行元：公益財団法人日本薬学会
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シナプスの病態 プリオン病
森 剛志, 坂口末廣
Clinical Neuroscience 28 ( 8 ) 906 - 908 2010年8月
記述言語：日本語 掲載種別：記事・総説・解説・論説等（学術雑誌） 出版者・発行元：中外医学社
講演・口頭発表等 【 表示 ／ 非表示 】
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ヒトグリオーマ細胞におけるプリオン感染動態の解析.
森剛志, 今村守一, 髙月英恵, 井口洋美, 大野美奈子, 新竜一郎
第68回日本ウイルス学会学術集会 2021年11月16日
開催年月日： 2021年11月16日 - 2021年11月18日
記述言語：日本語 会議種別：口頭発表（一般）
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スクレイピー感染ヒツジ脳には複数のプリオン株が混在している.
今村守一, 宮澤光太郎, 森剛志, 髙月英恵, 新竜一郎
九州微生物研究フォーラム2021 2021年9月11日
開催年月日： 2021年9月10日 - 2021年9月11日
記述言語：日本語 会議種別：口頭発表（一般）
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さまざまなリコンビナントプリオン蛋白を基質としたRT-QUIC法の反応性の比較解析.
森剛志, 今村守一, 高月英恵, 井口洋美, 大野美奈子, 新竜一郎
第67回日本ウイルス学会学術集会 (東京都江戸川区) 日本ウイルス学会
開催年月日： 2019年10月29日 - 2019年10月31日
記述言語：日本語 会議種別：ポスター発表
開催地：東京都江戸川区
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Isolation of hidden minor prion conformers from classical scrapie isolates in advanced protein misfolding cyclic amplification in the presence of arginine ethyl ester. 国際会議
Imamura M, Miyazawa K, Matsuura Y, Iwamaru Y, Kitamoto T, Mohri S, Takatsuki H, Mori T, Atarashi R
APPS2019 (Wako, Saitama) Asian Pacific Prion Symposium
開催年月日： 2019年10月3日 - 2019年10月4日
記述言語：英語 会議種別：ポスター発表
開催地：Wako, Saitama
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Pentosan polysulfate induces latent prion infection in Fukuoka-1 strain-infected cells. 国際会議
Takatsuki H, Mori T, Imamura M, Atarashi R
APPS2019 (Wako, Saitama) Asian Pacific Prion Symposium
開催年月日： 2019年10月3日 - 2019年10月4日
記述言語：英語 会議種別：ポスター発表
開催地：Wako, Saitama
科研費（文科省・学振・厚労省）獲得実績 【 表示 ／ 非表示 】
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脳・脊髄液以外の検体からの微量異常型プリオン蛋白検出法の開発
研究課題/領域番号：23K06946 2023年04月 - 2026年03月
独立行政法人日本学術振興会 科学研究費基金 基盤研究（C）
担当区分：研究代表者
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採取容易な検体を用いたクロイツフェルト・ヤコブ病早期診断法の確立
研究課題/領域番号：20K07807 2020年04月 - 2023年03月
科学研究費補助金 基盤研究(C)
担当区分：研究代表者
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クロイツフェルト・ヤコブ病のタイプを鑑別可能なコンビネーションQUIC法の構築
研究課題/領域番号：17K10022 2017年04月 - 2020年03月
科学研究費補助金 基盤研究(C)
担当区分：研究代表者
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プリオン高感度検出法を用いたヤコブ病タイプ鑑別診断法の開発とその分子機構の解明
研究課題/領域番号：26461507 2014年04月 - 2017年03月
科学研究費補助金 基盤研究(C)
担当区分：研究代表者