Selection of bone marrow ALDH bright cells for treatment of pulmonary fibrosis focusing on changes in bone marrow characteristics

2020 Fiscal Year Research-status Report

• Project/Area Number: 20K08519
• Research Institution: Hiroshima University
• Principal Investigator: 中島 拓 広島大学, 病院(医), 助教 (90643792)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: aldehyde dehydrogenase / 肺線維症 / 骨髄幹細胞 / 間質性肺炎

Outline of Annual Research Achievements

初年度となる本年度は、実験計画に従いまず健常マウスにおける骨髄細胞中のaldehyde dehydrogenase高発現細胞 (ALDHbr)の割合を確認することから着手した。ALDEFLUOR kitを用いた染色、およびフローサイトメーターによって検出された健常マウス骨髄におけるALDHbrは20%前後であった。
さらにCD45陽性 (血球系)およびCD45陰性(非血球系)にわけALDHbr陽性率の確認を試みた。CD45陰性細胞は、骨髄において約20%程度存在したが、これら骨髄CD45陰性細胞中にはALDHbrがほぼ存在しないことが判明した。
そのため、CD45のかわりに骨髄造血幹細胞の選別に利用されるLineage抗体 (CD4, CD8, Gr1, CD11b, CD45RO, Ter117など成熟した血球表面マーカーに対する抗体を混合したカクテル抗体)を用いて骨髄細胞を検証したところ、骨髄細胞の10%程度がLineage-、90%はLineage+であった。これらLineage+細胞およびLineage-細胞には一定のALDHbr分画が認められたことから、それぞれLineage+ALDHbrとLineage-ALDHbrを採取し肺線維症マウスに投与したところ、Lineage-ALDHbrを投与したマウスでのみ肺線維症の抑制効果が得られた。
現在はLineage-ALDHbrの肺線維症に対する有効性について再現性確認、および適切な投与タイミングの検証を行っている。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

CD45陰性細胞は、骨髄において約20%程度存在したが、これら骨髄CD45陰性細胞中にはALDHbrがほぼ存在しないことが明らかとなった。そのため、当初の計画である「CD45陽性ALDHbr vs. CD45陰性ALDHbr」の治療効果比較は実行できないことが判明した。
これら予想外の結果に対し、骨髄造血幹細胞の選別に利用されるLineage抗体 (CD4, CD8, Gr1, CD11b, CD45RO, Ter117など成熟した血球表面マーカーに対する抗体を混合したカクテル抗体)を用いて「Lineage+ALDHbr vs. Lineage-ALDHbr」の治療効果を比較することで当初の計画に沿った検討が可能であると考えている。

Strategy for Future Research Activity

「Lineage+ALDHbr vs. Lineage-ALDHbr」の治療効果を比較したところLineage-ALDHbrを投与したマウスでのみ肺線維症の抑制効果が得られている。まずこの結果の再現性確認および、最も治療効果が期待できる適切な投与タイミングの検証を行う。
上記と並行して、Lineage-ALDHbrの細胞分画をさらに細分化し、真に治療効果に寄与する細胞集団の同定を目指す。
また、ヒト臨床試験でみられるように疾患ドナー骨髄を用いた場合、すなわち疾患マウスである肺線維症マウスをドナーとした場合、治療効果が減弱ないしは消失するかどうかを検討する。くわえて、mCherryタンパクを核内に発現するmCherryマウスをドナーに用いてドナー細胞とレシピエント細胞の識別を可能とすることで、移植されたドナー細胞が肺内に定着するかどうか、定着しているとすればどのような細胞に分化しているか、などを検証する予定である。

Research Products

• [Journal Article] Correlations of forced oscillometric bronchodilator response with airway inflammation and disease duration in asthma. (2021)
  • Author(s): Higaki N, Iwamoto H, Yamaguchi K, Sakamoto S, Horimasu Y, Masuda T, Miyamoto S, Nakashima T, Ohshimo S, Fujitaka K, Hamada H, Hattori N.
  • Journal Title: Clin Respir J. Volume: 15 Pages: 48-55.
  • DOI: 10.1111/crj.13268.
  • Peer Reviewed
• [Journal Article] A machine-learning based approach to quantify fine crackles in the diagnosis of interstitial pneumonia: A proof-of-concept study. (2021)
  • Author(s): Horimasu Y, Ohshimo S, Yamaguchi K, Sakamoto S, Masuda T, Nakashima T, Miyamoto S, Iwamoto H, Fujitaka K, Hamada H, Sadamori T, Shime N, Hattori N.
  • Journal Title: Medicine (Baltimore). Volume: 100 Pages: e24738.
  • DOI: 10.1097/MD.0000000000024738.
  • Peer Reviewed / Open Access
• [Journal Article] Analysis of microRNA Expression in Liquid-Based Cytology Samples May Be Useful for Primary Lung Cancer Diagnosis. (2021)
  • Author(s): Araki Y, Arihiro K, Yamaguchi K, Sakamoto S, Horimasu Y, Masuda T, Miyamoto S, Nakashima T, Iwamoto H, Fujitaka K, Hamada H, Hattori N.
  • Journal Title: Am J Clin Pathol. Volume: N/A Pages: in press
  • DOI: 10.1093/ajcp/aqaa278.
  • Peer Reviewed
• [Journal Article] Clinical significance of BIM deletion polymorphism in chemoradiotherapy for non-small cell lung cancer. (2021)
  • Author(s): Wakabayashi Y, Masuda T, Fujitaka K, Nakashima T, Okumoto J, Shimoji K, Nishimura Y, Yamaguchi K, Sakamoto S, Horimasu Y, Miyamoto S, Iwamoto H, Ohshimo S, Hamada H, Hattori N.
  • Journal Title: Cancer Sci. Volume: 112 Pages: 369-379.
  • DOI: 10.1111/cas.14711.
  • Peer Reviewed / Open Access
• [Journal Article] Autoantibody Positivity Is a Risk Factor for Chemotherapy-induced Exacerbation of Interstitial Pneumonia in Lung Cancer. (2021)
  • Author(s): Ito N, Masuda T, Nakashima T, Nakao S, Yamaguchi K, Sakamoto S, Horimasu Y, Miyamoto S, Iwamoto H, Fujitaka K, Hamada H, Hattori N.
  • Journal Title: Anticancer Res. Volume: 41 Pages: 1497-1506.
  • DOI: 10.21873/anticanres.14908.
  • Peer Reviewed
• [Journal Article] Reduced endogenous secretory RAGE in blood and bronchoalveolar lavage fluid is associated with poor prognosis in idiopathic pulmonary fibrosis. (2020)
  • Author(s): Yamaguchi K, Iwamoto H, Mazur W, Miura S, Sakamoto S, Horimasu Y, Masuda T, Miyamoto S, Nakashima T, Ohshimo S, Fujitaka K, Hamada H, Hattori N.
  • Journal Title: Respir Res. Volume: 21 Pages: 145.
  • DOI: 10.1186/s12931-020-01410-3.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Coexisting TIF1γ-positive Primary Pulmonary Lymphoepithelioma-like Carcinoma and Anti-TIF1γ Antibody-positive Dermatomyositis. (2020)
  • Author(s): Nakanishi Y, Yamaguchi K, Yoshida Y, Sakamoto S, Horimasu Y, Masuda T, Nakashima T, Miyamoto S, Iwamoto H, Hirata S, Fujitaka K, Hamada H, Sugiyama E, Hattori N.
  • Journal Title: Intern Med. Volume: 59 Pages: 2553-2558.
  • DOI: 10.2169/internalmedicine.4702-20.
  • Peer Reviewed / Open Access
• [Journal Article] Extent of pulmonary fibrosis on high-resolution computed tomography is a prognostic factor in patients with pleuroparenchymal fibroelastosis. (2020)
  • Author(s): Namba M, Masuda T, Takao S, Terada H, Yamaguchi K, Sakamoto S, Horimasu Y, Miyamoto S, Nakashima T, Iwamoto H, Ohshimo S, Fujitaka K, Hamada H, Awai K, Hattori N.
  • Journal Title: Respir Investig. Volume: 58 Pages: 465-472.
  • DOI: 10.1016/j.resinv.2020.05.009.
  • Peer Reviewed
• [Journal Article] Serum high-mobility group box 1 as a predictive marker for cytotoxic chemotherapy-induced lung injury in patients with lung cancer and interstitial lung disease. (2020)
  • Author(s): Nakao S, Yamaguchi K, Iwamoto H, Sakamoto S, Horimasu Y, Masuda T, Miyamoto S, Nakashima T, Ohshimo S, Fujitaka K, Hamada H, Hattori N.
  • Journal Title: Respir Med. Volume: 172 Pages: 106131.
  • DOI: 10.1016/j.rmed.2020.106131.
  • Peer Reviewed
• [Journal Article] Association of Preexisting Interstitial Lung Abnormalities With Immune Checkpoint Inhibitor-Induced Interstitial Lung Disease Among Patients With Nonlung Cancers. (2020)
  • Author(s): Shimoji K, Masuda T, Yamaguchi K, Sakamoto S, Horimasu Y, Nakashima T, Miyamoto S, Iwamoto H, Fujitaka K, Hamada H, Takeno S, Hide M, Teishima J, Ohdan H, Hattori N.
  • Journal Title: JAMA Netw Open. Volume: 3 Pages: e2022906.
  • DOI: 10.1001/jamanetworkopen.2020.22906.
  • Peer Reviewed / Open Access
• [Presentation] 肺線維症マウスにおける骨髄性質の変化 (2020)
  • Author(s): 中島 拓
  • Organizer: 第60回日本呼吸器学会学術講演会
  • Invited
• [Presentation] EGFR gene mutation is not a significant risk factor in patients with EGFR mutated non-small cell lung cancer who receive best supportive care alone (2020)
  • Author(s): Masuda T, Wakabayashi Y, Shimoji K, Yamaguchi K, Sakamoto S, Nishimura Y, Horimasu Y, Nakashima T, Miyamoto S, Senoo T, Iwamoto H, Ohshimo S, Fujitaka K, Hamada H, Hattori N.
  • Organizer: 2020 ASCO Annual Meeting
  • Int'l Joint Research
• [Presentation] Clinical significance of BIM deletion polymorphism on chemoradiotherapy in patients with non-small cell lung cancer (2020)
  • Author(s): Wakabayashi Y, Masuda T, Nakashima T, Fujitaka K, Yamaguchi K, Sakamoto S, Nishimura Y, Horimasu Y, Miyamoto S, Senoo T, IwamotoH, Ohshimo S, Hamada H, Hattori N.
  • Organizer: 2020 ASCO Annual Meeting
  • Int'l Joint Research
• [Presentation] Pre-existing interstitial lung abnormalities are risk factors for immune checkpoint inhibitor-induced interstitial lung disease in non-NSCLC cancers. (2020)
  • Author(s): Shimoji K, Masuda T, Nakanishi Y, Yamaguchi K, Sakamoto S, Horimasu Y, Nakashima T, Miyamoto S, Iwamoto H, Fujitaka K, Hamada H, Hattori N.
  • Organizer: 2020 ASCO Annual Meeting
  • Int'l Joint Research
• [Presentation] Increased fraction of small round lymphocyte in BALF is a diagnostic indicator for sarcoidosis (2020)
  • Author(s): Horimasu Y, Yamaguchi K, Sakamoto S, Masuda T, Miyamoto S, Nakashima T, Iwamoto H, Fujitaka K, Hamada H, Hattori N.
  • Organizer: 30th International Congress of the European Respiratory Society
  • Int'l Joint Research
• [Presentation] Reduced endogenous secretory RAGE in blood and bronchoalveolar lavage fluid is associated with poor prognosis in idiopathic pulmonary fibrosis (2020)
  • Author(s): Yamaguchi K, Iwamoto H, Mazur W*, Miura S, Sakamoto S, Horimasu Y, Masuda T, Miyamoto S, Nakashima T, Ohshimo S, Fujitaka K, Hamada H, Hattori N.
  • Organizer: 30th International Congress of the European Respiratory Society
  • Int'l Joint Research
• [Presentation] IL-18 binding protein can be a prognostic biomarker for idiopathic pulmonary fibrosis (2020)
  • Author(s): Nakanishi Y, Horimasu Y, Yamaguchi K, Sakamoto S, Masuda T, Nakashima T, Miyamoto S, Iwamoto H, Ohshimo S, Fujitaka K, Hamada H, Hattori N.
  • Organizer: 30th International Congress of the European Respiratory Society
  • Int'l Joint Research
• [Presentation] Pivotal role of the HMGB1/sRAGE axis in drug-induced lung injury in advanced lung cancer with pre-existing interstitial lung disease (2020)
  • Author(s): Nakao S, Yamaguchi K, Sakamoto S, Horimasu Y, Masuda T, Miyamoto S, Nakashima T, Iwamoto H, Ohshimo S, Fujitaka K, Hamada H, Hattori N.
  • Organizer: 30th International Congress of the European Respiratory Society
  • Int'l Joint Research
• [Presentation] Correlations of forced oscillometric bronchodilator response with airway inflammation and disease duration in asthma (2020)
  • Author(s): Higaki N, Iwamoto H, Yamaguchi K, Sakamoto S, Horimasu Y, Masuda T, Miyamoto S, Nakashima T, Fujitaka K, Hamada H, Hattori N.
  • Organizer: JSA/WAO Joint Congress 2020
  • Int'l Joint Research
• [Presentation] Real-world study of prescription pattern of inhaled corticosteroids in adult asthma (2020)
  • Author(s): Nakanishi Y, Iwamoto H, Nakao S, Higaki N, Yamaguchi K, Sakamoto S, Horimasu Y, Masuda T, Matsumoto N, Nakashima T, Miyamoto S, Onari Y, Fujitaka K, Haruta Y, Hamada H, Hozawa S, Hattori N.
  • Organizer: JSA/WAO Joint Congress 2020
  • Int'l Joint Research
• [Presentation] Decline of lung function in adults with a history of childhood asthma (2020)
  • Author(s): Miura S, Iwamoto H, Omori K, Yamaguchi K, Sakamoto S, Horimasu Y, Masuda T, Miyamoto S, Nakashima T, Fujitaka K, Hamada H, Yokoyama A*, Hattori N.
  • Organizer: JSA/WAO Joint Congress 2020
  • Int'l Joint Research
• [Book] 呼吸器疾患最新の治療 2021-2022 (2021)
  • Author(s): 中島 拓，服部 登
  • Total Pages: 3
  • Publisher: 南江堂