Study on glidinig motility of Babesia bovis merozoite

• Project/Area Number: 16K08021
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Veterinary medical science
• Research Institution: Nagasaki University
• Principal Investigator: ASADA Masahito 長崎大学, 熱帯医学研究所, 助教 (40587028)
• Project Period (FY): 2016-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: バベシア / ウシ / 滑走運動 / ゲノム編集 / 原虫 / 遺伝子組換え / 運動性 / 寄生虫 / 赤血球侵入 / 遺伝子改変 / バベシア・ボビス / グライデオソーム

Outline of Final Research Achievements

Babesia bovis is an apicomplexan intraerythrocytic protozoan parasite that induces babesiosis in cattle after transmission by ticks. During specific stages of the apicomplexan parasite lifecycle, host cells are targeted for invasion using a unique, active process termed "gliding motility". We have reported gliding motility of B. bovis merozoites, however molecular mechanisms of gliding motility have not been elucidated. In this study, we have searched the genes related to the glideosome, and found 5 thrombospondin-related adhesive protein (TRAP) genes. The single gene knockout experiment revealed that four of the TRAP genes were dispensable. On the contrary, we failed to disrupt one of the TRAP gene. The glmS gene knock down system was developed for Babesia parasites and adopted to the TRAP gene, and we found the gene has an important role for intraerythrocytic stage of the parasite.

Academic Significance and Societal Importance of the Research Achievements

本研究では、バベシア原虫メロゾイト滑走運動の分子機序を明らかにするまでには至らなかったが、同原虫TRAP関連遺伝子の一つが赤内期の生存に重要な役割を果たすことを明らかにした。また、技術的な面からはバベシア原虫において初めて2種類の遺伝子ノックダウン法を確立したほか、CRISPR/Cas9法によるゲノム編集法を確立し、さらにFRETバイオセンサーを用いたイメージングに成功するなど顕著な進捗が見られた。遺伝子ノックダウン法やゲノム編集法は寄生体のゲノム機能解析に非常に有効なツールであるため、今後バベシア病研究の加速が見込まれる。

Research Products

• [Journal Article] Genome editing of Babesia bovis using the CRISPR/Cas9 system. (2019)
  • Author(s): Hakimi, H., Ishizaki, T., Kegawa, Y., Kaneko, O., Kawazu, S., and Asada, M.
  • Journal Title: mSphere Volume: 4(3) Issue: 3
  • DOI: 10.1128/msphere.00109-19
  • NAID: 120006988459
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Epidemiology, risk factors, and co-infection of vector-borne pathogens in goats from Sistan and Baluchestan province, Iran (2019)
  • Author(s): Hakimi H, Sarani A, Takeda M, Kaneko O, Asada M
  • Journal Title: PLoS One Volume: 14(6) Issue: 6 Pages: e0218609-e0218609
  • DOI: 10.1371/journal.pone.0218609
  • NAID: 120006987685
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The application of the HyPer fluorescent sensor in the real-time detection of H 2 O 2 in Babesia bovis merozoites in vitro (2018)
  • Author(s): Asada Masahito、Hakimi Hassan、Kawazu Shin-ichiro
  • Journal Title: Veterinary Parasitology Volume: 255 Pages: 78-82
  • DOI: 10.1016/j.vetpar.2018.03.016
  • Peer Reviewed / Open Access
• [Journal Article] Establishment of a stable transfection system for genetic manipulation of Babesia gibsoni (2018)
  • Author(s): Liu Mingming、Adjou Moumouni Paul Franck、Asada Masahito、Hakimi Hassan、Masatani Tatsunori、Vudriko Patrick、Lee Seung-Hun、Kawazu Shin-ichiro、Yamagishi Junya、Xuan Xuenan
  • Journal Title: Parasites & Vectors Volume: 11 Issue: 1 Pages: 260-260
  • DOI: 10.1186/s13071-018-2853-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identification and characterization of interchangeable cross-species functional promoters between Babesia gibsoni and Babesia bovis (2018)
  • Author(s): Liu M, Adjou Moumouni PF, Cao S, Asada M, Wang G, Gao Y, Guo H, Li J, Vudriko P, Efstratiou A, Ringo AE, Lee SH, Hakimi H, Masatani T, Sunaga F, Kawazu SI, Yamagishi J, Jia L, Inoue N, Xuan X
  • Journal Title: Ticks Tick Borne Dis Volume: 9 (2) Issue: 2 Pages: 330-333
  • DOI: 10.1016/j.ttbdis.2017.11.008
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Whole-genome assembly of Babesia ovata and comparative genomics between closely related pathogens. (2017)
  • Author(s): Yamagishi J, Asada M, Hakimi H, Tanaka TQ, Sugimoto C, Kawazu SI
  • Journal Title: BMC Genomics Volume: 18 (1) Issue: 1 Pages: 832-832
  • DOI: 10.1186/s12864-017-4230-4
  • NAID: 120006529641
  • Peer Reviewed / Open Access
• [Journal Article] Transient transfection of intraerythrocytic Babesia gibsoni using elongation factor-1 alpha promoter (2017)
  • Author(s): Liu M, Asada M, Cao S, Moumouni PFA, Vudriko P, Efstratiou A, Hakimi H, Masatani T, Sunaga F, Kawazu SI, Yamagishi J, Xuan X
  • Journal Title: Mol Biochem Parasitol Volume: 216 Pages: 56-59
  • DOI: 10.1016/j.molbiopara.2017.07.003
  • Peer Reviewed / Open Access
• [Journal Article] Detection and molecular characterization of Babesia, Theileria, and Hepatozoon species in hard ticks collected from Kagoshima, the southern region in Japan (2017)
  • Author(s): Masatani T, Hayashi K, Andoh M, Tateno M, Endo Y, Asada M, Kusakisako K, Tanaka T, Gokuden M, Hozumi N, Nakadohzono F, Matsuo T
  • Journal Title: Ticks and Tick Borne Diseases Volume: 印刷中 Issue: 4 Pages: 581-587
  • DOI: 10.1016/j.ttbdis.2017.03.007
  • Peer Reviewed / Open Access
• [Journal Article] Identification and functional analysis of a novel mitochondria-localized 2-Cys peroxiredoxin, BbTPx-2, from Babesia bovis (2016)
  • Author(s): Masatani T, Asada M, Hakimi H, Hayashi K, Yamagishi J, Kawazu S, Xuan X
  • Journal Title: Parasitology Research Volume: 115(8) Issue: 8 Pages: 3139-3145
  • DOI: 10.1007/s00436-016-5071-9
  • NAID: 120006390950
  • Peer Reviewed
• [Presentation] Proteomic approach leads to the identification of novel Babesia bovis proteins expressed on the surface of infected erythrocytes (2019)
  • Author(s): Hakimi H, Templeton TJ, Sakaguchi M, Kaneko O, Asada M
  • Organizer: 第88回日本寄生虫学会大会
• [Presentation] Babesia bovis ves1α expression is correlated with cytoadhesion of parasite-infected erythrocyte to the endothelial cells (2018)
  • Author(s): Asada M, Hakimi H, Yamagishi J, Sakaguchi M, Yahata K, Kawazu S, Kaneko O
  • Organizer: The 17th Awaji International Forum on Infection and Immunity
  • Int'l Joint Research
• [Presentation] Proteomic approach leads to the identification of nobel Babesia bovis proteins expressed on the surface of infected (2018)
  • Author(s): Hakimi H, Templeton TJ, Sakaguchi M, Kaneko O, Asada M
  • Organizer: The 17th Awaji International Forum on Infection and Immunity
  • Int'l Joint Research
• [Presentation] Proteomic approach leads to the identification of novel Babesia bovis proteins expressed on the surface of infected erythrocytes (2018)
  • Author(s): Hakimi H, Templeton TJ, Sakaguchi M, Kaneko O, Asada M
  • Organizer: 14th International Congress of Parasitology
  • Int'l Joint Research
• [Presentation] Babesia bovis ves1α expression is correlated with cytoadhesion of parasite-infected erythrocyte to the endothelial cells (2018)
  • Author(s): Asada M, Hakimi H, Yamagishi J, Sakaguchi M, Yahata K, Kawazu S, Kaneko O.
  • Organizer: 14th International Congress of Parasitology
  • Int'l Joint Research
• [Presentation] Babesia bovis感染赤血球の 宿主血管内皮細胞接着機構の解明 (2016)
  • Author(s): 麻田正仁、ハキミハッサン、山岸潤也、坂口美亜子、 矢幡一英、河津信一郎、金子修
  • Organizer: 第24回分子寄生虫学ワークショップ/第14回分子寄生虫・マラリア研究フォーラム
  • Place of Presentation: 帯広畜産大学(北海道・帯広市)
  • Year and Date: 2016-08-21