Mechanism of hemophagocytosis by abnormal activation of macrophages

• Project/Area Number: 19H03433
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 49010:Pathological biochemistry-related
• Research Institution: Kanazawa University
• Principal Investigator: Hanayama Rikinari 金沢大学, ナノ生命科学研究所, 教授 (40403191)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2022: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2021: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2020: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2019: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
• Keywords: 血球貪食 / マクロファージ / 炎症性疾患 / シグナルクロストーク / CAGE法 / サイトカインストーム / サイトカイン

Outline of Research at the Start

血球貪食症候群は、過度の炎症性刺激でマクロファージが異常活性化することにより、本来貪食対象ではない自己血球を生きたまま貪食するようになる疾患である。私達はこの疾患の発症機構を解明する為、培養細胞を用いて血球貪食を再現する実験系を樹立した。そこで本研究では、この培養細胞実験系を活用することにより、血球貪食の分子機構の全容を解明するとともに、それらの分子機構が実際のヒト患者やマウスモデルでの病態発症に関与するかを検討することで、将来の治療標的の同定を目指す。加えて、特定の炎症性刺激の組み合わせが、どのような相乗効果により血球貪食を誘発するのかを明らかにする。

Outline of Final Research Achievements

Macrophages phagocytose hemophagocytes upon triple stimulation with CpG DNA, IFN-γ, and anti-IL-10 receptor antibody.Therefore, by comparing gene expression in macrophages before and after stimulation, we identified 12 cell surface molecules whose expression was strongly upregulated by the triple stimulation. By evaluating inhibitory antibodies against each molecule and the effect of decreased expression in macrophages using CRISPR/Cas9 on hemophagocytosis ability, four molecules were identified as novel hemophagocytosis-related molecules. We also analyzed how the expression of these molecules is regulated by signal crosstalk between CpG DNA and IFN-γ using the CAGE method and found that the transcription start sites of seven specific genes are altered.

Academic Significance and Societal Importance of the Research Achievements

死細胞の貪食機構については数多くの研究がなされており、その生理的意義が明らかとなっているが、血球貪食の生理的意義は未だ不明である。この機構は重篤な炎症性疾患の病態だけではなく、より広範な「Cell-in-cell structures」として、胸腺ナース細胞による未熟Tリンパ球の取り込み・育成や、細胞の共食い（エントーシス）、リンパ球の経細胞内移動（trans-cellular migration）などの基盤としても生理的に重要である可能性がある。更に、この機構を制御することで、生きた癌細胞をマクロファージに貪食除去させる治療法が開発される可能性もあり、新たな研究展開への発展が期待される。

Research Products

• [Journal Article] EVs and Communication (2023)
  • Author(s): Yang Xi、Yoshida Takeshi、Hanayama Rikinari
  • Journal Title: Encyclopedia of Cell Biology. Bradshaw R. eds. (Elsevier) Volume: 2 Pages: 390-400
  • DOI: 10.1016/b978-0-12-821618-7.00151-6
  • ISBN: 9780128216248
  • Peer Reviewed
• [Journal Article] Nanoscopic Assessment of Anti-SARS-CoV-2 Spike Neutralizing Antibody Using High-Speed AFM. (2023)
  • Author(s): Keesiang Lim, Goro Nishide, Elma Sakinatus Sajidah, Tomoyoshi Yamano, Yujia Qiu, Takeshi Yoshida, Akiko Kobayashi, Masaharu Hazawa, Toshio Ando, Rikinari Hanayama, Richard W Wong
  • Journal Title: Nano letters Volume: 23 Issue: 2 Pages: 619-628
  • DOI: 10.1021/acs.nanolett.2c04270
  • Peer Reviewed / Open Access
• [Journal Article] TIM4-Affinity Methods Targeting Phosphatidylserine for Isolation or Detection of Extracellular Vesicles (2022)
  • Author(s): Yoshida Takeshi、Hanayama Rikinari
  • Journal Title: Methods in molecular biology (Clifton, N.J.) Volume: 2466 Pages: 23-36
  • DOI: 10.1007/978-1-0716-2176-9_2
  • ISBN: 9781071621752, 9781071621769
  • Peer Reviewed
• [Journal Article] Preventing SARS-CoV-2 Infection Using Anti-spike Nanobody-IFN-β Conjugated Exosomes. (2022)
  • Author(s): Xiabing Lyu, Shota Imai, Tomoyoshi Yamano, Rikinari Hanayama
  • Journal Title: Pharmaceutical research Volume: 40 Issue: 4 Pages: 927-935
  • DOI: 10.1007/s11095-022-03400-0
  • Peer Reviewed
• [Journal Article] Spatiotemporal tracking of small extracellular vesicle nanotopology in response to physicochemical stresses revealed by HS-AFM. (2022)
  • Author(s): Elma Sakinatus Sajidah, Keesiang Lim, Tomoyoshi Yamano, Goro Nishide, Yujia Qiu, Takeshi Yoshida, Hanbo Wang, Akiko Kobayashi, Masaharu Hazawa, Firli R P Dewi, Rikinari Hanayama, Toshio Ando, Richard W Wong
  • Journal Title: Journal of extracellular vesicles Volume: 11 Issue: 11 Pages: 12275-12275
  • DOI: 10.1002/jev2.12275
  • Peer Reviewed / Open Access
• [Journal Article] Extracellular vesicles contribute to the metabolism of transthyretin amyloid in hereditary transthyretin amyloidosis (2022)
  • Author(s): Yamaguchi H, Kawahara H, Kodera N, Kumaki A, Tada Y, Sakai K, Ono K, Yamada M, Hanayama R
  • Journal Title: Front Mol Biosci Volume: 9 Pages: 839917-839917
  • DOI: 10.3389/fmolb.2022.839917
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of LILRB3 and LILRA6 allelic variants in the Japanese population (2021)
  • Author(s): Hirayasu K, Sun J, Hasegawa G, Hashikawa Y, Hosomichi K, Tajima A, Tokunaga K, Ohashi J, Hanayama R.
  • Journal Title: Journal of Human Genetics Volume: 印刷中 Issue: 7 Pages: 739-748
  • DOI: 10.1038/s10038-021-00906-0
  • Peer Reviewed
• [Journal Article] Structural and mechanical characteristics of exosomes from osteosarcoma cells explored by 3D-atomic force microscopy (2021)
  • Author(s): Yurtsever Ayhan、Yoshida Takeshi、Badami Behjat Arash、Araki Yoshihiro、Hanayama Rikinari、Fukuma Takeshi
  • Journal Title: Nanoscale Volume: 13 Issue: 13 Pages: 6661-6677
  • DOI: 10.1039/d0nr09178b
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Osteosarcoma-Derived Small Extracellular Vesicles Enhance Tumor Metastasis and Suppress Osteoclastogenesis by miR-146a-5p (2021)
  • Author(s): Yoshihiro Araki, Hisaki Aiba, Takeshi Yoshida, Norio Yamamoto, Katsuhiro Hayashi, Akihiko Takeuchi, Shinji Miwa, Kentaro Igarashi, Tuan D. Nguyen, Kiyo-aki Ishii, Takayuki Nojima, Satoru Takahashi, Hideki Murakami, Hiroyuki Tsuchiya and Rikinari Hanayama
  • Journal Title: Frontiers in Oncology Volume: 11 Pages: 667109-667109
  • DOI: 10.3389/fonc.2021.667109
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identification of small compounds regulating the secretion of extracellular vesicles via a TIM4-affinity ELISA (2021)
  • Author(s): Ma Yunfei、Yoshida Takeshi、Matoba Kazutaka、Kida Katsuhiko、Shintani Rito、Piao Yingshi、Jin Jingchun、Nishino Taito、Hanayama Rikinari
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 13471-13471
  • DOI: 10.1038/s41598-021-92860-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Millisecond dynamic of SARS‐CoV‐2 spike and its interaction with ACE2 receptor and small extracellular vesicles (2021)
  • Author(s): Lim Keesiang、Nishide Goro、Yoshida Takeshi、Watanabe‐Nakayama Takahiro、Kobayashi Akiko、Hazawa Masaharu、Hanayama Rikinari、Ando Toshio、Wong Richard W.
  • Journal Title: Journal of Extracellular Vesicles Volume: 10 Issue: 14
  • DOI: 10.1002/jev2.12170
  • Peer Reviewed / Open Access
• [Journal Article] Cytoplasmic DNA accumulation preferentially triggers cell death of myeloid leukemia cells by interacting with intracellular DNA sensing pathway (2021)
  • Author(s): Baba Tomohisa、Yoshida Takeshi、Tanabe Yamato、Nishimura Tatsunori、Morishita Soji、Gotoh Noriko、Hirao Atsushi、Hanayama Rikinari、Mukaida Naofumi
  • Journal Title: Cell Death & Disease Volume: 12 Issue: 4 Pages: 322-322
  • DOI: 10.1038/s41419-021-03587-x
  • Peer Reviewed / Open Access
• [Journal Article] Glioma-derived extracellular vesicles promote tumor progression by conveying WT1 (2020)
  • Author(s): Tsutsui Taishi、Kawahara Hironori、Kimura Ryouken、Dong Yu、Jiapaer Shabierjiang、Sabit Hemragul、Zhang Jiakang、Yoshida Takeshi、Nakada Mitsutoshi、Hanayama Rikinari
  • Journal Title: Carcinogenesis Volume: 41 Issue: 9 Pages: 1238-1245
  • DOI: 10.1093/carcin/bgaa052
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Tumor‐secreted proliferin‐1 regulates adipogenesis and lipolysis in cachexia (2020)
  • Author(s): Nguyen Tuan Duc、Miyatake Yuji、Yoshida Takeshi、Kawahara Hironori、Hanayama Rikinari
  • Journal Title: International Journal of Cancer Volume: 148 Issue: 8 Pages: 1982-1992
  • DOI: 10.1002/ijc.33418
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] High-Speed AFM Reveals Molecular Dynamics of Human Influenza A Hemagglutinin and Its Interaction with Exosomes (2020)
  • Author(s): Lim Keesiang、Kodera Noriyuki、Wang Hanbo、Mohamed Mahmoud Shaaban、Hazawa Masaharu、Kobayashi Akiko、Yoshida Takeshi、Hanayama Rikinari、Yano Seiji、Ando Toshio、Wong Richard W.
  • Journal Title: Nano Letters Volume: 20 Issue: 9 Pages: 6320-6328
  • DOI: 10.1021/acs.nanolett.0c01755
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Transient IGF-1R inhibition combined with osimertinib eradicates AXL-low expressing EGFR mutated lung cancer (2020)
  • Author(s): Wang Rong、Yamada Tadaaki、Kita Kenji、Taniguchi Hirokazu、Arai Sachiko、Fukuda Koji、Terashima Minoru、Ishimura Akihiko、Nishiyama Akihiro、Tanimoto Azusa、Takeuchi Shinji、Ohtsubo Koshiro、Wang Wei、Suzuki Takeshi、Yano Seiji et. al.
  • Journal Title: Nature Communications Volume: 11 Issue: 1 Pages: 4607-4607
  • DOI: 10.1038/s41467-020-18442-4
  • NAID: 120007186801
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Emerging roles of extracellular vesicles in physiology and disease (2020)
  • Author(s): Hanayama Rikinari
  • Journal Title: The Journal of Biochemistry Volume: 169 Issue: 2 Pages: 135-138
  • DOI: 10.1093/jb/mvaa138
  • Peer Reviewed / Open Access
• [Journal Article] Novel properties of myoferlin in glucose metabolism via pathways involving modulation of adipose functions (2020)
  • Author(s): Nozato Y, Takami Y, Yamamoto K, Nagasawa M, Nozato S, Imaizumi Y, Takeshita H, Wang C, Ito Y, Takeda S, Takeya Y, Sugimoto K, Nakagami H, Hanayama R, Rakugi H.
  • Journal Title: FASEB J. Volume: 34 Issue: 2 Pages: 2792-2811
  • DOI: 10.1096/fj.201901539rr
  • Peer Reviewed / Open Access
• [Journal Article] Development of a Method That Delivers Drugs to Enveloped Viruses (2019)
  • Author(s): Bahrini Insaf、Hanayama Rikinari
  • Journal Title: Biological and Pharmaceutical Bulletin Volume: 42 Issue: 6 Pages: 977-981
  • DOI: 10.1248/bpb.b18-01000
  • NAID: 130007657691
  • ISSN: 0918-6158, 1347-5215
  • Year and Date: 2019-06-01
  • Open Access
• [Journal Article] Aire-expressing ILC3-like cells in the lymph node display potent APC features (2019)
  • Author(s): Yamano Tomoyoshi、Dobes Jan、Voboril Matous、Steinert Madlen、Brabec Tomas、Zietara Natalia、Dobesova Martina、Ohnmacht Caspar、Laan Martti、Peterson Part、Benes Vladimir、Sedlacek Radislav、Hanayama Rikinari、Kolar Michal、Klein Ludger、Filipp Dominik
  • Journal Title: The Journal of Experimental Medicine Volume: 印刷中 Issue: 5 Pages: 1027-1037
  • DOI: 10.1084/jem.20181430
  • Peer Reviewed
• [Journal Article] Serum milk fat globule-EGF factor 8 (MFG-E8) as a diagnostic and prognostic biomarker in patients with hepatocellular carcinoma (2019)
  • Author(s): Shimagaki Tomonari、Yoshio Sachiyo、Kawai Hironari、Sakamoto Yuzuru、Doi Hiroyoshi、Matsuda Michitaka、Mori Taizo、Osawa Yosuke、Fukai Moto、Yoshida Takeshi、Ma Yunfei、Akita Tomoyuki、Tanaka Junko、Taketomi Akinobu、Hanayama Rikinari、Yoshizumi Tomoharu、Mori Masaki、Kanto Tatsuya
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 15788-15788
  • DOI: 10.1038/s41598-019-52356-6
  • Open Access
• [Presentation] Co-evolution of bacteria and paired immune receptors in humans (2023)
  • Author(s): 平安恒幸，華山力成
  • Organizer: 第96回 日本細菌学術総会
• [Presentation] LILRファミリーを介した宿主と細菌の共進化 (2022)
  • Author(s): 長谷川玄, 平安恒幸，華山力成
  • Organizer: 第12回 東日本研究医養成コンソーシアム
• [Presentation] LILRB3 and LILRA6 recognize bacteria in an allele-specific manner (2022)
  • Author(s): 長谷川玄, 平安恒幸，華山力成
  • Organizer: 第51回 日本免疫学会学術総会
• [Presentation] 自己炎症疾患の発症機序とエクソソームの役割 (2021)
  • Author(s): 華山力成
  • Organizer: 第1回 日本耳鼻咽喉科免疫アレルギー感染症学会
  • Invited
• [Presentation] マクロファージによる自己炎症応答とエクソソームによるその制御 (2019)
  • Author(s): 華山力成
  • Organizer: 第92回 日本生化学会大会
  • Invited
• [Book] 実験医学増刊 シン・マクロファージ（佐藤荘 編，羊土社） (2022)
  • Author(s): 平安恒幸，華山力成
  • Total Pages: 7
  • Publisher: 羊土社