Characterization of Aspergillus derived extracellular vesicles and application to a diagnostics

• Project/Area Number: 17K10040
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Infectious disease medicine
• Research Institution: National Institute of Infectious Diseases
• Principal Investigator: Miyazaki Yoshitsugu 国立感染症研究所, 真菌部, 部長 (00311861)
• Co-Investigator(Kenkyū-buntansha): 山越 智 国立感染症研究所, 真菌部, 室長 (00212283)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 分泌小胞 / RNA / 膜タンパク質 / ガラクトマンナン / アスペルギルス / 病原性真菌診断系

Outline of Final Research Achievements

The research object is to develop a new diagnostic method for invasive aspergillosis by detecting extracellular vesicles (EVs) on the basis of our presumption that EVs of Aspergillus fumigatus are secreted into blood and urine. We have analyzed proteins and RNAs in EVs prepared by ultracentrifugation. Finally, we have constructed detection systems of RNA by RT-PCR, but not reached to establish a concentration method of EVs by an antibody against membrane proteins of EVs.

Academic Significance and Societal Importance of the Research Achievements

本研究においてアスペルギルス症の主な原因菌であるAspergillus fumigatusでも他の真菌と同様に分泌小胞が確認され、さらにRNAの解析より様々な小分子RNAが同定された。人をはじめとする高等生物の分泌小胞の1つであるエクソソームでは、含まれる小分子RNAの種類が病気を反映すること、また、それがエクソソームを介して他の細胞に伝達され病気に関係することが分かってきており、アスペルギルス症においても、真菌の分泌小胞のRNAが、病態を反映するだけでなく病原性にも関与することも考えられ、本研究で得られた成果は、今後の真菌症の基礎および臨床研究に寄与するものと考えられる。

Research Products

• [Journal Article] Selection of Oral Antifungals for Initial Maintenance Therapy in Chronic Pulmonary Aspergillosis: A Longitudinal Analysis. (2020)
  • Author(s): Masato Tashiro,Takahiro Takazono,Tomomi Saijo,Kazuko Yamamoto,Yoshifumi Imamura,Taiga Miyazaki,Hiroshi Kakeya,Tsunehiro Ando,Kenji Ogawa,Kazuma Kishi,Issei Tokimatsu,Yuta Hayashi,Satoru Fujiuchi,Katsunori Yanagihara,Yoshitsugu Miyazaki,Kiyoshi Ichihara,Hiroshi Mukae,Shigeru Kohno,Koichi Izumikawa]
  • Journal Title: Clinical infectious diseases : an official publication of the Infectious Diseases Society of America Volume: 70 Issue: 5 Pages: 835-842
  • DOI: 10.1093/cid/ciz287
  • Peer Reviewed / Open Access
• [Journal Article] Echinocandins versus non-echinocandins for empirical antifungal therapy in patients with hematological disease with febrile neutropenia: A systematic review and meta-analysis. (2020)
  • Author(s): Chizuru Yamashita,Yoshio Takesue,Kazuaki Matsumoto,Kazuhiro Ikegame,Yuki Enoki,Motoi Uchino,Taiga Miyazaki,Koichi Izumikawa,Tohru Takada,Keiji Okinaka,Takashi Ueda,Yoshitsugu Miyazaki,Toshihiko Mayumi]
  • Journal Title: Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy Volume: 26 Issue: 6 Pages: 596-603
  • DOI: 10.1016/j.jiac.2020.01.015
  • Peer Reviewed / Open Access
• [Journal Article] Chronic Nodular Pulmonary Aspergillosis in a Patient with Rheumatoid Arthritis (2019)
  • Author(s): Yuhei Ito,Motoaki Tanigawa,Tamiko Takemura,Keisuke Iwamoto,Akina Nigi,Hajime Sasano,Hidetoshi Itani,Shigeto Kondo,Toshiya Tokui,Shigeki Nakamura,Yoshitsugu Miyazaki]
  • Journal Title: Internal Medicine Volume: 58 Issue: 7 Pages: 979-984
  • DOI: 10.2169/internalmedicine.1918-18
  • NAID: 130007622191
  • ISSN: 0918-2918, 1349-7235
  • Year and Date: 2019-04-01
  • Peer Reviewed / Open Access
• [Journal Article] Identification of a Novel Variant Form of <i>Aspergillus fumigatus</i> CalC and Generation of Anti-CalC Monoclonal Antibodies (2019)
  • Author(s): Takatsuka Shogo、Inukai Tatsuya、Kawakubo Shun、Umeyama Takashi、Abe Masahiro、Ueno Keigo、Hoshino Yasutaka、Kinjo Yuki、Miyazaki Yoshitsugu、Yamagoe Satoshi
  • Journal Title: Medical Mycology Journal Volume: 60 Issue: 1 Pages: 11-16
  • DOI: 10.3314/mmj.18-00013
  • NAID: 130007605409
  • ISSN: 2185-6486, 2186-165X
  • Peer Reviewed / Open Access
• [Journal Article] Disseminated histoplasmosis from a calcified lung nodule after long-term corticosteroid therapy in an elderly Japanese patient (2019)
  • Author(s): Kobayashi Keigo、Asakura Takanori、Kawada Ichiro、Hasegawa Hanako、Chubachi Shotaro、Ohara Kentaro、Kuramoto Junko、Sugiura Hiroaki、Fujishima Seitaro、Iwata Satoshi、Umeyama Takashi、Katano Harutaka、Uwamino Yoshifumi、Miyazaki Yoshitsugu、Kamei Katsuhiko、Hasegawa Naoki、Betsuyaku Tomoko
  • Journal Title: Medicine Volume: 98 Issue: 17 Pages: e15264-e15264
  • DOI: 10.1097/md.0000000000015264
  • Peer Reviewed / Open Access
• [Journal Article] Gastric mucormycosis complicated by a gastropleural fistula: A case report and review of the literature. (2019)
  • Author(s): Tomohisa Uchida,Momoko Okamoto,Keita Fujikawa,Daisuke Yoshikawa,Akinari Mizokami,Tomo Mihara,Akira Kondo,Kazuo Ohba,Kazuhiro Kurohama,Masahiro Nakashima,Ichiro Sekine,Shigeki Nakamura,Yoshitsugu Miyazaki,Atsushi Kawakami]
  • Journal Title: Medicine Volume: 98 Issue: 48 Pages: e18142-e18142
  • DOI: 10.1097/md.0000000000018142
  • Peer Reviewed / Open Access
• [Journal Article] The incidence of endophthalmitis or macular involvement and the necessity of a routine ophthalmic examination in patients with candidemia. (2019)
  • Author(s): Ueda T, Takesue Y, Tokimatsu I, Miyazaki T, Nakada-Motokawa N, Nagao M, Nakajima K, Mikamo H, Yamagishi Y, Kasahara K, Yoshihara S, Ukimura A, Yoshida K, Yoshinaga N, Izumi M, Kakeya H, Yamada K, Kawamura H, Endou K, Yamanaka K, Yoshioka M, Amino K, Ikeuchi H, Uchino M, Miyazaki Y.
  • Journal Title: PLoS One. Volume: 14 Issue: 5 Pages: e0216956-e0216956
  • DOI: 10.1371/journal.pone.0216956
  • NAID: 120006719416
  • Peer Reviewed / Open Access
• [Journal Article] Neutrophil-mediated antifungal activity against highly virulent Cryptococcus gattii strain R265 (2019)
  • Author(s): Ueno Keigo、Yanagihara Nao、Otani Yoshiko、Shimizu Kiminori、Kinjo Yuki、Miyazaki Yoshitsugu
  • Journal Title: Medical Mycology Volume: 印刷中 Issue: 8 Pages: 1046-1054
  • DOI: 10.1093/mmy/myy153
  • Peer Reviewed
• [Journal Article] Efficacy of T-2307, a novel arylamidine, against ocular complications of disseminated candidiasis in mice (2019)
  • Author(s): Abe Masahiro、Nakamura Shigeki、Kinjo Yuki、Masuyama Yuka、Mitsuyama Junichi、Kaku Mitsuo、Miyazaki Yoshitsugu
  • Journal Title: Journal of Antimicrobial Chemotherapy Volume: 74 Issue: 5 Pages: 1327-1332
  • DOI: 10.1093/jac/dkz020
  • Peer Reviewed
• [Journal Article] A dendritic cell-based systemic vaccine induces long-lived lung-resident memory Th17 cells and ameliorates pulmonary mycosis (2018)
  • Author(s): Ueno Keigo、Urai Makoto、Sadamoto Sota、Shinozaki Minoru、Takatsuka Shogo、Abe Masahiro、Otani Yoshiko、Yanagihara Nao、Shimizu Kiminori、Iwakura Yoichiro、Shibuya Kazutoshi、Miyazaki Yoshitsugu、Kinjo Yuki
  • Journal Title: Mucosal Immunology Volume: 12 Issue: 1 Pages: 265-276
  • DOI: 10.1038/s41385-018-0094-4
  • Peer Reviewed / Open Access
• [Journal Article] CRISPR/Cas9 Genome Editing To Demonstrate the Contribution of Cyp51A Gly138Ser to Azole Resistance in Aspergillus fumigatus (2018)
  • Author(s): Umeyama Takashi、Hayashi Yuta、Shimosaka Hisaki、Inukai Tatsuya、Yamagoe Satoshi、Takatsuka Shogo、Hoshino Yasutaka、Nagi Minoru、Nakamura Shigeki、Kamei Katsuhiko、Ogawa Kenji、Miyazaki Yoshitsugu
  • Journal Title: Antimicrobial Agents and Chemotherapy Volume: 62 Issue: 9
  • DOI: 10.1128/aac.00894-18
  • Peer Reviewed
• [Journal Article] Differences in Ocular Complications Between Candida albicans and Non-albicans Candida Infection Analyzed by Epidemiology and a Mouse Ocular Candidiasis Model (2018)
  • Author(s): Abe Masahiro、Kinjo Yuki、Ueno Keigo、Takatsuka Shogo、Nakamura Shigeki、Ogura Sho、Kimura Muneyoshi、Araoka Hideki、Sadamoto Sota、Shinozaki Minoru、Shibuya Kazutoshi、Yoneyama Akiko、Kaku Mitsuo、Miyazaki Yoshitsugu
  • Journal Title: Frontiers in Microbiology Volume: 9 Pages: 1266-1266
  • DOI: 10.3389/fmicb.2018.02477
  • Peer Reviewed / Open Access
• [Journal Article] Micafungin Breakthrough Fungemia in Patients with Hematological Disorders (2018)
  • Author(s): Kimura Muneyosh, et al
  • Journal Title: Antimicrobial Agents and Chemotherapy Volume: 62 Issue: 5
  • DOI: 10.1128/aac.02183-17
  • Peer Reviewed
• [Journal Article] Intraspecies variation in the efficacy of adjunctive recombinant interferon-γ therapy against cryptococcal meningoencephalitis in mice. (2017)
  • Author(s): Ikeda-Dantsuji Y, Nakamura S, Ohno H, Inukai T, Nagi M, Ueno K, Umeyama T, Kinjo Y, Yamagoe S, Shibuya K, Miyazaki Y
  • Journal Title: Medical mycology Volume: 56 Issue: 3 Pages: 382-386
  • DOI: 10.1093/mmy/myx063
  • Peer Reviewed
• [Journal Article] A Giant Dermatophyte Abscess Caused by <I>Trichophyton rubrum</I> in an Immunocompromised Patient (2017)
  • Author(s): 1.Toyoki M, Hase M, Hirasawa Y, Umeyama T, Miyazaki Y, Kano R, Sugita T, Hiruma M, Kodama Y, Ikeda S.
  • Journal Title: Medical Mycology Journal Volume: 58 Issue: 2 Pages: E63-E66
  • DOI: 10.3314/mmj.17-00003
  • NAID: 130006906288
  • ISSN: 2185-6486, 2186-165X
  • Peer Reviewed
• [Journal Article] Immunization with Antigen-Pulsed Dendritic Cells Against Highly Virulent Cryptococcus gattii Infection: Analysis of Cytokine-Producing T Cells (2017)
  • Author(s): Ueno Keigo、Urai Makoto、Takatsuka Shogo、Abe Masahiro、Miyazaki Yoshitsugu、Kinjo Yuki
  • Journal Title: Methods Mol Biol. Volume: 1625 Pages: 327-339
  • DOI: 10.1007/978-1-4939-7104-6_22
  • ISBN: 9781493971039, 9781493971046
  • Peer Reviewed
• [Journal Article] Successful Treatment of Pulmonary Mucormycosis Caused by Cunninghamella bertholletiae with High-Dose Liposomal Amphotericin B (10 mg/kg/day) Followed by a Lobectomy in Cord Blood Transplant Recipients. (2017)
  • Author(s): Ota H, Yamamoto H, Kimura M, Araoka H, Fujii T, Umeyama T, Ohno H, Miyazaki Y, Kaji D, Taya Y, Nishida A, Ishiwata K, Tsuji M, Takagi S, Asano-Mori Y, Yamamoto G, Uchida N, Izutsu K, Masuoka K, Wake A, Yoneyama A, Makino S, Taniguchi S.
  • Journal Title: Mycopathologia Volume: 61 Issue: 9-10 Pages: 847-853
  • DOI: 10.1007/s11046-017-0149-1
  • Peer Reviewed
• [Journal Article] Identification of a novel Rhizopus-specific antigen by screening with a signal sequence trap and evaluation as a possible diagnostic marker of mucormycosis (2017)
  • Author(s): Sato K., Oinuma K., Niki M., Yamagoe S., Miyazaki Y., Asai K., Yamada K., Hirata K., Kaneko Y. and Kakeya H.
  • Journal Title: Medical Mycology Volume: 印刷中 Issue: 7 Pages: 713-719
  • DOI: 10.1093/mmy/myw146
  • NAID: 120006533842
  • Peer Reviewed
• [Journal Article] Mycotic pseudoaneurysm of a pulmonary artery branch caused by Cladosporium (2017)
  • Author(s): Sunagawa Keishin、Uchino Yoshihito、Ishimoto Shinichirou、Nakamura Shigeki、Honma Taku、Nakanishi Yoko、Hatta Yoshihiro、Miyazaki Yoshitsugu、Sakurai Hiroyuki、Hao Hiroyuki、Sugitani Masahiko
  • Journal Title: Pathology International Volume: 68 Issue: 1 Pages: 47-52
  • DOI: 10.1111/pin.12615
  • Peer Reviewed
• [Presentation] 2016～2018年3年間実施したBiosafety level 3対応が必要な真菌検査について (2020)
  • Author(s): 壇辻百合香、星野泰隆、名木 稔、阿部雅弘、上野圭吾、中山靖子、橋本ゆき、東 祥嗣、犬飼達也、高塚翔吾、梅山 隆、山越 智、宮﨑義継
  • Organizer: 第31回日本臨床微生物学会総会・学術集会
• [Presentation] CRISPR / Cas9 genome editing technology to verify the contribution of known mutations in the cyp51A gene and its promoter to azole resistance in Aspergillus fumigatus. (2020)
  • Author(s): Takashi Umeyama, Tatsuya Inukai, Madoka Tateno, Satoshi Yamagoe, Shogo Takatsuka, Yasutaka Hoshino, Keiko Ishino, Yoshitsugu Miyazaki
  • Organizer: 9th Advanecs Against Aspergillosis and Mucormycosis
  • Int'l Joint Research
• [Presentation] Hmg1 mutation conferring multi-azole resistance in Aspergillus fumigatus (2020)
  • Author(s): Teppei Arai, Takashi Umeyama, Tatsuya Inukai, Akira Watanabe, Yoshitsugu Miyazaki, Katsuhiko Kamei
  • Organizer: 9th Advanecs Against Aspergillosis and Mucormycosis
  • Int'l Joint Research
• [Presentation] 注意すべき糸状菌 (2020)
  • Author(s): 壇辻百合香、星野泰隆、名木稔、阿部雅広、上野圭吾、中山靖子、橋本ゆき、東祥嗣、犬飼達也、高塚翔吾、梅山隆、山越智、宮崎義継
  • Organizer: 第31回 日本臨床微生物学会総会・学術集会
• [Presentation] Aspergillus fumigatusの血清存在下での増殖機構 (2020)
  • Author(s): 犬飼達也、梅山 隆、山越 智、宮崎義継
  • Organizer: 第93回日本細菌学会総会
• [Presentation] クリプトコックス症に対する新規経鼻ワクチンの開発とその作用機構: 免疫賦活性に優れたワクチン株の作製 (2020)
  • Author(s): 上野 圭吾, 柳原 尚, 清水 公徳, 山越 智, 宮﨑 義継
  • Organizer: 第93回 日本細菌学会総会
• [Presentation] Aspergillus fumigatusの細胞外小胞の解析 (2019)
  • Author(s): 山越 智、犬飼達也、高塚翔吾、星野泰隆、岡本圭祐、上野圭吾、梅山 隆、宮﨑義継
  • Organizer: 第13回アスペルギルス研究会
• [Presentation] Aspergillus fumigatusアゾール耐性株にみられるCyp51A SNPSのゲノム編集による直接検証 (2019)
  • Author(s): 梅山 隆、犬飼達也、山越 智、髙塚翔吾、星野泰隆、宮﨑義継
  • Organizer: 第63回日本医真菌学会総会・学術集会
• [Presentation] Aspergillus fumigatus B11b破壊株の増殖を抑制する血清中の成分の同定 (2019)
  • Author(s): 犬飼達也、星野泰隆、浦井 誠、梅山 隆、高塚翔吾、山越 智、東 祥嗣、阿部雅広、名木 稔、中村茂樹、宮崎義継
  • Organizer: 第63回日本医真菌学会総会・学術集会
• [Presentation] インフルエンザ続発性肺アスペルギルス症モデルの確立と重症化メカニズムに関する研究 (2019)
  • Author(s): 高塚翔吾, 林崎浩史, 犬飼達也, 山越智, 梅山隆, 星野泰隆, 上野圭吾, 宮崎義継
  • Organizer: 第63回日本医真菌学会総会・学術集会
• [Presentation] Genome editing technology to elucidate the antifungal resistance mechanisms in filamentous fungi (2019)
  • Author(s): Takashi Umeyama, Tatsuya Inukai, Madoka Tateno, Satoshi Yamagoe, Shogo Takatsuka, Yasutaka Hoshino, Yoshitsugu Miyazaki
  • Organizer: The Asia Pacific Society for Medical Mycology Congress 2019
  • Int'l Joint Research
• [Presentation] CRISPR/Cas9ゲノム編集技術によるAspergillus fumigatusボリコナゾール耐性変異の検証 (2018)
  • Author(s): 梅山隆, 犬飼達也, 山越智, 名木稔, 中村茂樹, 小川賢二, 宮﨑義継,
  • Organizer: 第92回日本感染症学会総会・学術講演会 第66回日本化学療法学会学術集会 合同学会
• [Presentation] アスペルギルス・フミガタスのアゾール系抗真菌薬耐性株の解析 (2018)
  • Author(s): 星野泰隆, 宮崎義継
  • Organizer: 第62回 日本医真菌学会総会
• [Presentation] Aspergillus fumigatusの血清存在下の生育に必須な因子の同定と病原性解析 (2018)
  • Author(s): 犬飼達也, 梅山 隆, 山越智, 宇田晶彦, 星野泰隆, 高塚翔吾, 阿部雅広, 栃木直文, 篠崎稔, 渋谷和俊, 名木稔, 壇辻百合香, 中村茂樹, 宮崎義継
  • Organizer: 第12回アスペルギルス研究会
• [Presentation] パラフィン包埋試料を用いた真菌DNA検出状況に関する検討 (2018)
  • Author(s): 壇辻百合香, 中村茂樹, 東祥嗣, 阿部雅広, 上野圭吾, 梅山隆, 山越智, 宮﨑義継
  • Organizer: 第65回日本化学療法学会東日本支部総会合同学会
• [Presentation] Mutations in the cyp51A gene of an azole resistant clinical isolate of Aspergillus fumigatus: direct proof using Cas9/CRISPR genome editing (2018)
  • Author(s): T. Umeyama, Y. Hayashi, H. Shimosaka, T. Inukai, S. Yamagoe, M. Nagi, S. Nakamura, K. Ogawa, Y. Miyazaki
  • Organizer: 8th Advanecs Against Aspergillosis
  • Int'l Joint Research
• [Presentation] Uptake of exogenous sterol promotes antifungal resistance in Candida glabrata (2018)
  • Author(s): 名木 稔，田辺公一，上野圭吾、犬飼達也、中村茂樹，梅山 隆，山越 智，宮﨑義継
  • Organizer: 第91回 日本細菌学会総会
• [Presentation] Exogenous sterol uptake: A potential cause of azole resistance (2018)
  • Author(s): 田辺 公一，名木 稔，犬飼 達也，中山 浩伸，梅山 隆，山越 智，中村 茂樹，宮﨑義継
  • Organizer: 第91回 日本細菌学会総会
• [Presentation] 病原性真菌Cryptococcus gattii の新規感染機構: 莢膜多糖による抗原被覆作用について (2018)
  • Author(s): 大谷淑子, 上野圭吾, 柳原 尚, 浦井 誠, 清水公徳, 宮﨑義継, 金城雄樹
  • Organizer: 第91回 日本細菌学会総会
• [Presentation] 好中球は高病原性真菌Cryptococcus gattii の増殖を抑制するか? (2018)
  • Author(s): 上野圭吾, 柳原 尚, 大谷淑子, 清水公徳, 宮﨑義継, 金城雄樹
  • Organizer: 第91回 日本細菌学会総会
• [Presentation] 侵襲性真菌症治療の新しい試み (2017)
  • Author(s): 宮﨑義継
  • Organizer: 第22回日本神経感染症学会総会・学術大会
  • Invited
• [Presentation] 真菌感染症の新しい診断法 (2017)
  • Author(s): 中村茂樹，中山靖子，壇辻百合香, 名木 稔, 浦井誠, 梅山 隆，山越 智，宮﨑義継
  • Organizer: 第91回 日本感染症学会総会・第65回日本化学療法学会総会
• [Presentation] クリプトコックスの莢膜欠損株は何故病原性が低いのか？: 環境適応と免疫回避機構における莢膜合成の意義 (2017)
  • Author(s): 上野圭吾, 金城雄樹, 浦井 誠, 宮﨑義継
  • Organizer: 第91回 日本感染症学会総会・第65回日本化学療法学会総会
• [Presentation] クラリスロマイシンによる高病原性Cryptococcus gattiiの莢膜形成抑制作用に関する検討 (2017)
  • Author(s): 中村茂樹，壇辻百合香, 犬飼達也, 名木 稔, 浦井誠, 梅山 隆，山越 智，宮﨑義継
  • Organizer: 第91回 日本感染症学会総会・第65回日本化学療法学会総会
• [Presentation] アスペルギルス症 (2017)
  • Author(s): 山越 智, 高塚翔吾, 梅山 隆, 犬飼達也, 名木 稔, 中村茂樹, 宮﨑義継
  • Organizer: 第61回 日本医真菌学会総会
• [Presentation] 病原性および鉄欠乏環境におけるCandida glabrataのマイトファジーの役割 (2017)
  • Author(s): 名木 稔，田辺公一，上野圭吾,中山浩伸、中村茂樹，梅山 隆，山越 智，宮﨑義継
  • Organizer: 第61回 日本医真菌学会総会
• [Presentation] 肺常在性記憶型Th17細胞は、高病原性肺クリプトコックス症を制御する (2017)
  • Author(s): 上野圭吾, 金城雄樹, 定本聡太, 篠崎 稔, 阿部雅広, 澁谷和俊, 宮﨑義継
  • Organizer: 第61回 日本医真菌学会総会