The elucidation of molecular basis of the diversity and transmissibility in abnormal prion protein formation

• Project/Area Number: 15H04269
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Nerve anatomy/Neuropathology
• Research Institution: University of Miyazaki (2016-2018); Nagasaki University (2015)
• Principal Investigator: Atarashi Ryuichiro 宮崎大学, 医学部, 教授 (90452846)
• Research Collaborator: IMAMURA morikazu
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥14,430,000 (Direct Cost: ¥11,100,000、Indirect Cost: ¥3,330,000); Fiscal Year 2017: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2016: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2015: ¥7,670,000 (Direct Cost: ¥5,900,000、Indirect Cost: ¥1,770,000)
• Keywords: プリオン病 / プリオン株 / PrP / タンパク単独仮説 / 神経変性疾患 / リコンビナントタンパク / プリオンタンパク

Outline of Final Research Achievements

Accumulating evidence suggests that the continuous conversion of normal form of prion protein (PrPC) into abnormal form of PrP (PrPSc) in affected hosts plays a central role of the pathogenesis of prion diseases. However, the molecular basis of the diversity and transmissibility in PrPSc formation has not been well understood. Using the protein misfolding cyclic amplification (PMCA) technique, we found that some cofactors such as heparin sulfate or its analog heparin greatly enhanced the formation of PrPSc in vitro. Furthermore, we developed an advanced PMCA system using E-coli derived recombinant PrP as a substrate. We currently examined the diversity and transmissibility of the advanced PMCA products.

Academic Significance and Societal Importance of the Research Achievements

補助的因子を加えた試験管内異常化反応法により、これまで効率の低かったプリオン株（非典型牛海綿状脳症や慢性消耗病等）の高効率な変換が可能となった。またその補助的因子を加えることにより、大腸菌由来リコンビナントPrP（recPrP）を反応基質とした試験管内PrP異常化反応系において、そのプリオン株特有の産物を生成させることが初めて可能となった。これらの結果は、プリオン病の病態機構の理解を大きく進展させたと考えている。

Research Products

• [Journal Article] Development of radioiodinated acridine derivatives for in vivo imaging of prion deposits in the brain. (2017)
  • Author(s): Kawasaki M, Fuchigami T, Kobashi N, Nakagaki T, Sano K, Atarashi R, Yoshida S, Haratake M, Nishida N, Nakayama M
  • Journal Title: Bioorganic & Medicinal Chemistry. Volume: 25 Issue: 3 Pages: 1085-1093
  • DOI: 10.1016/j.bmc.2016.12.020
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Real-Time Quaking-Induced Conversion for Diagnosis of Prion Disease. (2017)
  • Author(s): Satoh K, Atarashi R, Nishida N.
  • Journal Title: Methods Mol Biol. Volume: 1658 Pages: 305-310
  • DOI: 10.1007/978-1-4939-7244-9_21
  • ISBN: 9781493972425, 9781493972449
  • Peer Reviewed
• [Journal Article] A direct assessment of human prion adhered to steel wire using real-time quaking-induced conversion. (2016)
  • Author(s): Mori T, Atarashi R, Furukawa K, Takatsuki H, Satoh K, Sano K, Nakagaki T, Ishibashi D, Ichimiya K, Hamada M, Nakayama T, Nishida N.
  • Journal Title: Sci Rep. Volume: 6 Issue: 1 Pages: 24993-24993
  • DOI: 10.1038/srep24993
  • Peer Reviewed / Open Access
• [Journal Article] CSF RT-QuIC is a robust and reliable test for sporadic CJD: An international study. (2016)
  • Author(s): McGuire LI, Poleggi A, Poggiolini I, Suardi S, Grznarova K, Shi S, de Vil B, Sarros S, Satoh K, Cheng K, Cramm M, Fairfoul G, Schmitz M, Zerr I, Cras P, Equestre M, Tagliavini F, Atarashi R, Knox D, Collins S, Haik S, Parchi P, Pocchiari M, Green A.
  • Journal Title: Ann Neurol. Volume: 未定 Issue: 1 Pages: 160-165
  • DOI: 10.1002/ana.24679
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structure-Based Drug Discovery for Prion Disease Using a Novel Binding Simulation. (2016)
  • Author(s): Ishibashi D, Nakagaki T, Ishikawa T, Atarashi R, Watanabe K, Cruz FA, Hamada T, Nishida N.
  • Journal Title: EBioMedicine. Volume: 9 Pages: 238-249
  • DOI: 10.1016/j.ebiom.2016.06.010
  • NAID: 120006987627
  • Peer Reviewed / Open Access
• [Journal Article] The real-time quaking-induced conversion assay for detection of human prion disease and study of other protein misfolding diseases. (2016)
  • Author(s): Schmitz M, Cramm M, Llorens F, Muller-Cramm D, Collins S, Atarashi R, Satoh K, Orru CD, Groveman BR, Zafar S, Schulz-Schaeffer WJ, Caughey B, Zerr I.
  • Journal Title: Nat Protoc. Volume: 11 Issue: 11 Pages: 2233-2242
  • DOI: 10.1038/nprot.2016.120
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Prion-Seeding Activity Is widely Distributed in Tissues of Sporadic Creutzfeldt-Jakob Disease Patients. (2016)
  • Author(s): Takatsuki H, Fuse T, Nakagaki T, Mori T, Mihara B, Takao M, Iwasaki Y, Yoshida M, Murayama S, Atarashi R, Nishida N, Satoh K.
  • Journal Title: EBioMedicine. Volume: 12 Pages: 150-155
  • DOI: 10.1016/j.ebiom.2016.08.033
  • NAID: 120006987618
  • Peer Reviewed / Open Access
• [Journal Article] Characterisation of radioiodinated flavonoid derivatives for SPECT imaging of cerebral prion deposits. (2015)
  • Author(s): Fuchigami T, Yamashita Y, Kawasaki M, Ogawa A, Haratake M, Atarashi R, Sano K, Nakagaki T, Ubagai K, Ono M, Yoshida S, Nishida N, Nakayama M.
  • Journal Title: Sci Rep. Volume: 5 Issue: 1 Pages: 18440-18440
  • DOI: 10.1038/srep18440
  • NAID: 120006987233
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Strain-Dependent Effect of Macroautophagy on Abnormally Folded Prion Protein Degradation in Infected Neuronal Cells. (2015)
  • Author(s): Ishibashi D, Homma T, Nakagaki T, Fuse T, Sano K, Takatsuki H, Atarashi R, Nishida N.
  • Journal Title: PLoS One. Volume: 10 Issue: 9 Pages: 0137958-0137958
  • DOI: 10.1371/journal.pone.0137958
  • NAID: 120006986825
  • Peer Reviewed / Open Access
• [Journal Article] Structural conservation of prion strain specificities in recombinant prion protein fibrils in real-time quaking-induced conversion. (2015)
  • Author(s): Sano K, Atarashi R, Nishida N.
  • Journal Title: Prion. Volume: 9 Issue: 4 Pages: 237-243
  • DOI: 10.1080/19336896.2015.1062201
  • Peer Reviewed / Open Access
• [Journal Article] Rapid and Quantitative Assay of Amyloid-Seeding Activity in Human Brains Affected with Prion Diseases. (2015)
  • Author(s): Takatsuki H, Satoh K, Sano K, Fuse T, Nakagaki T, Mori T, Ishibashi D, Mihara B, Takao M, Iwasaki Y, Yoshida M, Atarashi R, Nishida N.
  • Journal Title: PLoS One Volume: 10 Issue: 6 Pages: 0126930-0126930
  • DOI: 10.1371/journal.pone.0126930
  • NAID: 120006986998
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Ubiquitin-specific protease 14 modulates degradation of cellular prion protein. (2015)
  • Author(s): Homma T, Ishibashi D, Nakagaki T, Fuse T, Mori T, Satoh K, Atarashi R, Nishida N.
  • Journal Title: Sci Rep. Volume: 5 Issue: 1 Pages: 11028-11028
  • DOI: 10.1038/srep11028
  • Peer Reviewed / Open Access
• [Presentation] 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. (2017)
  • Author(s): 新竜一郎, 今村守一, 森剛志, 高月英恵, 岩丸祥史
  • Organizer: 日本薬学会第138年会(シンポジウム)
  • Invited
• [Presentation] Spontaneously-generated baculovirus-derived abnormal recombinant prion protein has infectivity. (2017)
  • Author(s): Imamura M, Tabeta N, Iwamaru Y, Yokoyama T, Murayama Y, Mori T, Takatsuki H, Atarashi R.
  • Organizer: 第65回日本ウイルス学会学術集会
• [Presentation] Wire-QuIC reaction can detect abnormal human prion seeds from contaminated stainless steel-wire (2016)
  • Author(s): Mori T, Atarashi R, Satoh K,Ishibashi D, Nakagaki T, Nishida N.
  • Organizer: 第64回日本ウイルス学会学術集会
  • Place of Presentation: 札幌、日本
  • Year and Date: 2016-10-23
• [Presentation] "Wire-QuIC": a new detection sysmte of human prion (2016)
  • Author(s): Mori T, Atarashi R, Takatsuki H, Satoh K, Nakagaki T, Ishibashi D, Nishida N.
  • Organizer: Prion 2016
  • Place of Presentation: 東京、日本
  • Year and Date: 2016-05-10
  • Int'l Joint Research
• [Presentation] Diagnostic evaluation of real-time quaking-induced conversion for the detection of human prion diseases. (2015)
  • Author(s): Ryuichiro Atarashi
  • Organizer: Asian Pacific Prion Symposium 2015
  • Place of Presentation: 石川県立音楽堂(石川県・金沢市）
  • Year and Date: 2015-09-04
  • Int'l Joint Research / Invited
• [Remarks] 宮崎大学ニュースリソース
  • URL: http://www.miyazaki-u.ac.jp/newsrelease/edu-info/alprenolol.html