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2020 Fiscal Year Research-status Report

鉄過剰状態における造血幹細胞内鉄代謝制御の解明-アナモルシンを中心に-

Research Project

Project/Area Number 19K08861
Research InstitutionOsaka University

Principal Investigator

柴山 浩彦  大阪大学, 医学系研究科, 招聘教授 (60346202)

Co-Investigator(Kenkyū-buntansha) 野崎 健司  大阪大学, 医学部附属病院, 医員 (00836413)
横田 貴史  大阪大学, 医学系研究科, 講師 (60403200)
Project Period (FY) 2019-04-01 – 2022-03-31
Keywordsアナモルシン / 鉄・硫黄クラスター / アポトーシス / 遺伝子欠損マウス / 条件付き遺伝子欠損マウス / 造血幹細胞 / RNA-Seq
Outline of Annual Research Achievements

本研究では、申請者らがクローニングした鉄・硫黄クラスター形成に重要な役割を有する抗アポトーシス分子アナモルシン(AM)の造血幹細胞における機能と鉄過剰状態がAMの機能におよぼす影響についての解析をおこなっている。申請者らは、AMコンディショナルノックアウトマウス(AM flox/floxマウス)を作製することで、in vivoにおいて、種々の臓器、細胞のAMを欠損させることを可能としている。本研究では、MX-Creリコンビナーゼトランスジェニック(MX-Cre-Tg)マウスとAM flox/floxマウスを交配した MX-Cre-Tg/AM flox/floxマウスの造血幹細胞に焦点をあてた研究をおこなっている。MX-Cre-Tg/AM flox/floxマウスにPIPCを投与することで、造血幹細胞のAMを欠損させることができる。実際、PIPCを投与したところ、一週間程度で、一部のマウスは死亡したが、投与後も生存するマウスが存在した。これらの生存したマウスの血球数を経時的に測定したところ、PIPC投与後に一過性に白血球、赤血球の減少を認めるものの、その後、血球数の回復が確認された。また、同時に、AM遺伝子のノックダウンが生じている割合を確認するため、LOX-P配列の前後に設定したプライマー配列を用いたReal time PCRでAM遺伝子の欠損割合を定量したところ、約10%程度は、AM遺伝子をノックダウンできずに残存していることが判明した。PIPC投与後も生存したマウスにおいては、これらの造血幹細胞が、再度、増殖し血球回復したものと考えられた。以上の結果から、MX-Cre-Tg/AM flox/floxマウスにPIPCを投与し、AMを欠損した造血幹細胞のみを純化し抽出したmRNAを用いて網羅的にシークエンスをおこない、AM欠損による遺伝子の発現変化を検討する。

Current Status of Research Progress
Current Status of Research Progress

3: Progress in research has been slightly delayed.

Reason

準備実験では、MX-Cre-Tg/AM flox/floxマウスにPIPCを投与したところ、ほとんどのマウスが死亡したため、MX-Cre-Tg/AM flox/floxマウスから分離した造血幹細胞を正常マウスに移植し、キメラ状態のマウスを作成し、さらに、それらのマウスを鉄過剰状態にしたあとで、PIPCを投与する必要があると考えていたが、MX-Cre-Tg/AM flox/floxマウスにPIPCを投与しても、多くのマウスは、一過性に血球減少をきたすのみで、死亡まで至らないため、移植実験をおこなわずとも、AM欠損造血幹細胞を得ることが可能であることが判明した。しかし、PIPC投与後に、AMを欠損した造血幹細胞は、アポトーシスをおこし、その細胞数は減少するため、PIPC投与後に造血幹細胞を純化するタイミングについて、詳細に検討する必要があった。しかし、コロナ感染症が広がり、研究者が研究室や動物舎に来ることが制限される日があり、必要な数のMX-Cre-Tg/AM flox/floxマウスをそろえることが困難で、予定通りに実験を進めることができなかった。

Strategy for Future Research Activity

今後は、最初にMX-Cre-Tg/AM flox/floxマウスの数を増やし実験に足るだけの遺伝子改変マウスを準備する。そのマウスにPIPCを投与し、造血幹細胞において、最もAM遺伝子が欠損できるタイミングを、経時的に検体を採取することで明らかにする必要がある。その結果から、AMを最も欠損できるタイミングにおいて、純化した造血幹細胞からmRNAを抽出し、mRNA Seqをおこなうことで、AMを欠損させた造血幹細胞での遺伝子発現の変化を網羅的に解析する。変化がみられた遺伝子の中で、細胞の生存・増殖・アポトーシスに関わる遺伝子に着目する。現在までに、他の細胞(Bリンパ球)において、AM遺伝子を欠損させた際に、P38 MAPK遺伝子の発現低下がみられたというデータを得ており、造血幹細胞においても、P38 MAPK遺伝子の発現に変化がみられるかどうかに着目している。その後は、さらに、鉄過剰状態としたMX-Cre-Tg/AM flox/floxマウスにPIPCを投与し、血球数の動態に影響が認められるかどうか、さらに、その際に、AMを欠損させた造血幹細胞での遺伝子発現の変化をmRNA Seqをおこなうことで、同様に解析する予定である。

  • Research Products

    (33 results)

All 2021 2020

All Journal Article (17 results) (of which Peer Reviewed: 16 results,  Open Access: 15 results) Presentation (16 results) (of which Int'l Joint Research: 3 results)

  • [Journal Article] Subcutaneous delivery of daratumumab in Japanese patients with relapsed/refractory multiple myeloma.2021

    • Author(s)
      Shibayama H, Matsumoto M, Kosugi H, Shibayama K, Yamazaki H, Iida S.
    • Journal Title

      Int J Hematol

      Volume: 113 Pages: 112-121

    • DOI

      10.1007/s12185-020-02985-9.

    • Peer Reviewed / Open Access
  • [Journal Article] Ectonucleotidase CD39 is highly expressed on ATLL cells and is responsible for their immunosuppressive function.2021

    • Author(s)
      Nagate Y, Ezoe S, Fujita J, Okuzakis D, Motooka D, Ishibashi T, Ichii M, Tanimura A, Kurashige M, Morii E, Fukushima T, Suehiro Y, Yokota T, Shibayama H, Oritani K, Kanakura Y.
    • Journal Title

      Leukemia

      Volume: 35 Pages: 107-118

    • DOI

      10.1038/s41375-020-0788-y.

    • Peer Reviewed / Open Access
  • [Journal Article] Graft-versus-host disease develops in mice transplanted with lymphocyte-depleted bone marrow cells from signal-transducing adaptor protein-2 transgenic mice.2021

    • Author(s)
      Saito H, Ichii M, Toda J, Kitai Y, Muromoto R, Kashiwakura JI, Saitoh K, Tanimura A, Yokota T, Shibayama H, Matsuda T, Oritani K, Kanakura Y, Hosen N.
    • Journal Title

      Biochem Biophys Res Commun

      Volume: 537 Pages: 118-124

    • DOI

      10.1016/j.bbrc.2020.12.080.

    • Peer Reviewed / Open Access
  • [Journal Article] Signal-transducing adaptor protein-2 delays recovery of B lineage lymphocytes during hematopoietic stress.2021

    • Author(s)
      Ichii M, Oritani K, Toda J, Saito H, Shi H, Shibayama H, Motooka D, Kitai Y, Muromoto R, Kashiwakura JI, Saitoh K, Okuzaki D, Matsuda T, Kanakura Y.
    • Journal Title

      Haematologica

      Volume: 106 Pages: 424-436

    • DOI

      10.3324/haematol.2019.225573.

    • Peer Reviewed / Open Access
  • [Journal Article] Once-weekly vs. twice-weekly carfilzomib dosing in a subgroup of Japanese relapsed and refractory multiple myeloma patients from a randomized phase 3 trial (A.R.R.O.W.) and comparison with ENDEAVOR.2020

    • Author(s)
      Takezako N, Shibayama H, Handa H, Hagiwara S, Ozaki S, Suzuki K, Kosugi H, Ri M, Sugiura I, Choi I, Miyamoto T, Iida S.
    • Journal Title

      Int J Hematol

      Volume: 113 Pages: 219-230

    • DOI

      10.1007/s12185-020-03013-6.

    • Peer Reviewed / Open Access
  • [Journal Article] Isatuximab monotherapy in relapsed/refractory multiple myeloma: A Japanese, multicenter, Phase 1/2, safety and efficacy study.2020

    • Author(s)
      Sunami K, Suzuki K, Ri M, Matsumoto M, Shimazaki C, Asaoku H, Shibayama H, Ishizawa K, Takamatsu H, Ikeda T, Maruyama D, Kaneko H, Uchiyama M, Kiguchi T, Iyama S, Murakami H, Takahashi K, Tada K, S, Guillemin-Paveau H, Iida S.
    • Journal Title

      Cancer Sci

      Volume: 111 Pages: 4526-4539

    • DOI

      10.1111/cas.14657.

    • Peer Reviewed / Open Access
  • [Journal Article] Retrospective multi-center study of Adolescent and Young Adult (AYA) Multiple Myeloma in Kansai Myeloma Forum registry.2020

    • Author(s)
      Nakaya A, Kohara T, Shibayama H, Onda Y, Kanda J, Kaneko H, Kosugi S, Ishikawa J, Tanaka H, Fuchida SI, Shimura Y, Ohta K, Shimazaki C, Hino M, Takaori-Kondo A, Kuroda J, Matsumura I, Kanakura Y, Nomura S; Kansai Myeloma Forum Investigators.
    • Journal Title

      Int J Hematol

      Volume: 112 Pages: 435-438

    • DOI

      10.1007/s12185-020-02996-6.

    • Peer Reviewed / Open Access
  • [Journal Article] Whole-exome sequencing identified mutational profile of a case with T-cell chronic lymphocytic leukemia.2020

    • Author(s)
      Nozaki K, Yokota T, Itotagawa E, Tsutsumi K, Kusakabe S, Morikawa Y, Fujita J, Fukushima K, Maeda T, Shibayama H, Hosen N, Kumanogo A, Kanakura Y.
    • Journal Title

      Clin Case Rep

      Volume: 8 Pages: 2251-2254

    • DOI

      10.1002/ccr3.3149.

    • Peer Reviewed / Open Access
  • [Journal Article] Retrospective analysis of plasmacytoma in Kansai Myeloma Forum Registry.2020

    • Author(s)
      Nakaya A, Tanaka H, Yagi H, Ohta K, Shibayama H, Kohara T, Kanda J, Shindo M, Shimura Y, Kosugi S, Kaneko H, Fuchida SI, Uoshima N, Shimazaki C, Hino M, Kuroda J, Kanakura Y, Takaori-Kondo A, Nomura S, Matsumura I; Kansai Myeloma Forum Investigators.
    • Journal Title

      Int J Hematol

      Volume: 112 Pages: 666-673

    • DOI

      10.1007/s12185-020-02961-3.

  • [Journal Article] Signal-transducing adapter protein-1 is required for maintenance of leukemic stem cells in CML.2020

    • Author(s)
      Toda J, Ichii M, Oritani K, Shibayama H, Tanimura A, Saito H, Yokota T, Motooka D, Okuzaki D, Kitai Y, Muromoto R, Kashiwakura JI, Matsuda T, Hosen N, Kanakura Y.
    • Journal Title

      Oncogene

      Volume: 39 Pages: 5601-5615

    • DOI

      10.1038/s41388-020-01387-9.

    • Peer Reviewed / Open Access
  • [Journal Article] A Multicenter, Open-Label, Phase II Study of Tirabrutinib (ONO/GS-4059) in Patients With Waldenstrom's Macroglobulinemia.2020

    • Author(s)
      Sekiguchi N, Rai S, Munakata W, Suzuki K, Handa H, Shibayama H, Endo T, Terui Y, Iwaki N, Fukuhara N, Tatetsu H, Iida S, Ishikawa T, Shiibashi R, Izutsu K.
    • Journal Title

      Cancer Sci

      Volume: 111 Pages: 3327-3337

    • DOI

      10.1111/cas.14561.

    • Peer Reviewed / Open Access
  • [Journal Article] Clinical Utility of Target Capture-Based Panel Sequencing in Hematological Malignancies: A Multicenter Feasibility Study.2020

    • Author(s)
      Yasuda T, Sanada M, Ri M, Handa H, Ishida T, Shibayama H, Abe M, Kataoka K, Ogawa S, Iida S, Tomita A, Horibe K.
    • Journal Title

      Cancer Sci

      Volume: 111 Pages: 3367-3378

    • DOI

      10.1111/cas.14552.

    • Peer Reviewed / Open Access
  • [Journal Article] A Multi-Center Retrospective Analysis of Patients With Relapsed/Refractory Follicular Lymphoma After Third-Line Chemotherapy.2020

    • Author(s)
      Fuji S, Tada Y, Nozaki K, Saito H, Ozawa T, Kida T, Kosugi S, Sugahara H, Ikeda H, Hashimoto K, Karasuno T, Ueda S, Ishikawa J, Shibayama H.
    • Journal Title

      Ann Hematol

      Volume: 99 Pages: 2133-2139

    • DOI

      10.1007/s00277-020-04126-y.

    • Peer Reviewed / Open Access
  • [Journal Article] Pretreatment serum soluble interleukin-2 receptor level predicts survival in patients with newly diagnosed follicular lymphoma.2020

    • Author(s)
      Nozaki K, Sugahara H, Ueda S, Ishikawa J, Fuji S, Masaie H, Tada Y, Karasuno T, Iida M, Mitsui H, Kamae T, Saito N, Moriyama Y, Kawakami M, Kato R, Nakae Y, Kida T, Kosugi S, Nakagawa M, Uchida Y, Azenishi Y, Sakaniwa R, Kitamura T, Shibayama H.
    • Journal Title

      Leukemia and Lymphoma

      Volume: 61 Pages: 2113-2121

    • DOI

      10.1080/10428194.2020.1759054.

    • Peer Reviewed
  • [Journal Article] Limbic encephalitis with antibodies to N-methyl-D-aspartate (NMDA)-type glutamate receptor after allogeneic transplantation.2020

    • Author(s)
      Toda J, Maeda T, Akuta K, Kusakabe S, Ueda T, Fujita J, Shibayama H, Oritani K, Takahashi Y, Kanakura Y.
    • Journal Title

      Int J Hematol

      Volume: 112 Pages: 254-257

    • DOI

      10.1007/s12185-020-02859-0.

    • Peer Reviewed / Open Access
  • [Journal Article] Enterococcus: A predictor of ravaged microbiota and poor prognosis after allogeneic hematopoietic stem cell transplantation.2020

    • Author(s)
      Kusakabe S, Fukushima K, Yokota T, Hino A, Fujita J, Motooka D, Nakamura S, Shibayama H, Kanakura Y.
    • Journal Title

      Biol Blood Marrow Transplant

      Volume: 26 Pages: 1028-1033

    • DOI

      10.1016/j.bbmt.2020.01.019.

    • Peer Reviewed / Open Access
  • [Journal Article] Daratumumab, lenalidomide, and dexamethasone in Japanese patients with transplant-ineligible newly diagnosed multiple myeloma: a phase 1b study.2020

    • Author(s)
      Takamatsu H, Iida S, Shibayama H, Shibayama K, Yamazaki H, Suzuki K.
    • Journal Title

      Int J Hematol

      Volume: 115 Pages: 692-701

    • DOI

      10.1007/s12185-020-02825-w.

    • Peer Reviewed / Open Access
  • [Presentation] Genomic Analysis of FLT3 Mutations in a Comprehensive NGS Multicenter Study of AML: HM-Screen-Japan 01.2020

    • Author(s)
      Kentaro Fukushima, SungGi Chi, , Hirohiko Shibayama, , Naoko Hosono, Kenichi Miyamoto, Yosuke Minami
    • Organizer
      The American Society of Hematology 62nd Annual Meeting
    • Int'l Joint Research
  • [Presentation] ASXL1 Mutations Predict a Poor Response to Darbepoetin Alfa in Anemic Patients with Low-Risk MDS: A Multicenter, Phase II Study.2020

    • Author(s)
      Hitoshi Hanamoto, Yasuyoshi Morita, Motoshi Ichikawa, Yasuhito Nannya, Hirohiko Shibayama, Yoshinobu Maeda, Tomoko Hata, Toshihiro Miyamoto, Hiroshi Kawabata, Kazuto Takeuchi, Hiroko Tanaka, Junji Kishimoto, Satoru Miyano, Itaru Matsumura, Seishi Ogawa, Koichi Akashi, Yuzuru Kanakura, Kinuko Mitani
    • Organizer
      The American Society of Hematology 62nd Annual Meeting
    • Int'l Joint Research
  • [Presentation] The Phase 3 TOURMALINE-MM2 Trial: Oral Ixazomib, Lenalidomide, and Dexamethasone (IRd) Vs Placebo-Rd for Transplant-Ineligible Patients with Newly Diagnosed Multiple Myeloma (NDMM).2020

    • Author(s)
      Thierry Facon, Christopher P. Nizar Bahlis, Fritz Offner, Darrell J White, Lotfi Benboubker, Sophie Rigaudeau, Philippe Rodon, Sung-Soo Yoon, Kenshi Suzuki, Hirohiko Shibayama, Xiaoquan Zhang, Godwin Yung, Robert M. Rifkin, Philippe Moreau, Sagar Lonial, Shaji K. Kumar, Paul G. Richardson, Vincent Rajkumar
    • Organizer
      The American Society of Hematology 62nd Annual Meeting
    • Int'l Joint Research
  • [Presentation] Polatuzumab vedotin with bendamustine and rituximab in r/ r DLBCL (P-DRIVE): A phase 2 study in Japan2020

    • Author(s)
      Hirohiko Shibayama,Shinya Rai,Koji Izutsu,Motoko Yamaguchi, Jun Takizawa,Misato Hashizume,Yasuhito Terui
    • Organizer
      第82回日本血液学会学術集会
  • [Presentation] ASXL1 mutations predict a poor response to darbepoetin alfa in anemic patients with low-risk MDS2020

    • Author(s)
      Hirohiko Shibayama, Yasuyoshi Morita, Motoshi Ichikawa, Yasuhito Nannya, Hitoshi Hanamoto, Yoshinobu Maeda, Tomoko Hata, Toshihiro Miyamoto, Hiroshi Kawabata,Kazuto Takeuchi,Hiroko Tanaka,Junji Kishimoto,Satoru Miyano,Itaru Matsumura,Seishi Ogawa, Koichi Akashi, Yuzuru Kanakura, Kinuko Mitani
    • Organizer
      第82回日本血液学会学術集会
  • [Presentation] Patient demographics in a prospective observational study of plasma cell neoplasms in Japan2020

    • Author(s)
      Hirohiko Shibayama,Mitsuhiro Itagaki,Hiroshi Handa,Akihiro Yokoyama,Morio Sawamura,Toshiki Saito,Itaru Matsumura,Koichi Akashi,Shinsuke Iida
    • Organizer
      第82回日本血液学会学術集会
  • [Presentation] A multi-center retrospective analysis of relapsed/refractory FL after 3rd-line chemotherapy2020

    • Author(s)
      Shigeo Fuji,Yuma Tada,Kenji Nozaki,Hideaki Saito, Takayuki Ozawa,Toru Kida,Satoru Kosugi,Hiroyuki Sugahara, Hirokazu Ikeda,Koji Hashimoto,Takahiro Karasuno,Shuji Ueda,Jun Ishikawa,Hirohiko Shibayama
    • Organizer
      第82回日本血液学会学術集会
  • [Presentation] Multi-color analysis of immune response of myeloma patients under lenalidomide maintenance2020

    • Author(s)
      Kenji Nozaki,Yuki Fujioka,Jun Ishikawa,Masato Iida,Masaru Shibata,Satoru Kosugi,Hirohiko Shibayama
    • Organizer
      第82回日本血液学会学術集会
  • [Presentation] Induction therapies for transplant-ineligible NDMM: retrospective analysis of 547 cases from KMF2020

    • Author(s)
      Yuji Shimura,Hirohiko Shibayama,Aya Nakaya,Ryosuke Yamamura,Kazunori Imada,Masayuki Hino,Akifumi Takaori,Itaru Matsumura,Yuzuru Kanakura,Shosaku Nomura
    • Organizer
      第82回日本血液学会学術集会
  • [Presentation] Daratumumab/Len/Dex (D-Rd) in non-transplant NDMM: Update of MMY1006 (Japanese patients) and MAIA2020

    • Author(s)
      Saad Z. Usmani,Hirohiko Shibayama,Kenshi Suzuki,Shinsuke Iida,Thierry Facon,Nizar J. Bahlis,Hiroshi Yamazaki,Jianping Wang,Rian Van Rampelbergh,Hiroyuki Takamatsu
    • Organizer
      第82回日本血液学会学術集会
  • [Presentation] Subanalysis of FLT3 mutations in comprehensive NGS multicenter study of AML: HM-SCREEN-Japan 012020

    • Author(s)
      Kentaro Fukushima, Hirohiko Shibayama, Kenichi Miyamoto, Naoko Hosono, Takahiro Yamauchi, Nobuhiko Yamauchi, Yosuke Minami
    • Organizer
      第82回日本血液学会学術集会
  • [Presentation] PET-CT after bendamustine + rituximab for follicular lymphoma in first relapse can predict PFS2020

    • Author(s)
      Miyuki Okura,Koji Kato,Junji Suzumiya,Koji Izutsu,Momoko Nishikori, Hirohiko Shibayama,Yoshinobu Maeda,Kenichi Yoshimura,Ukihide Tateishi,Toshihiro Miyamoto,Jun Ishikawa,Shinya Rai,Tsutomu Takahashi,Koichi Akashi,Yuzuru Kanakura
    • Organizer
      第82回日本血液学会学術集会
  • [Presentation] Carfilzomib and dexamethasone (Kd) with/without daratumumab (D): subgroup analysis in Asian patients2020

    • Author(s)
      Kenshi Suzuki,Chang-Ki Min,Kihyun Kim,Je-Jung Lee,Hirohiko Shibayama,Po-Shen Ko,Shang-Yi Huang,Sin-Syue Li,Bifeng Ding,Monica Khurana,Seasea Gao,Shinsuke Iida
    • Organizer
      第82回日本血液学会学術集会
  • [Presentation] A phase 1 study of acalabrutinib in Japanese patients with B-cell malignancies2020

    • Author(s)
      Hirokazu Nagai,Kiyoshi Ando,Daisuke Ennishi,Hirohiko Shibayama,Junji Suzumiya,Kazuhito Yamamoto,Satoshi Ichikawa,Koji Kato,Kyoya Kumagai,Priti Patel,Sakura Iizumi,Nobuya Hayashi,Hisashi Kawasumi,Koji Izutsu
    • Organizer
      第82回日本血液学会学術集会
  • [Presentation] Autonomous TGFβ signaling promotes phenotypic variations of human AML cells2020

    • Author(s)
      Yasuhiro Shingai,Takafumi Yokota,Ritsuko Nakai,Takayuki Ozawa, Tomoaki Ueda,Yukiko Doi,Tomohiko Ishibashi,Michiko Ichii, Hirohiko Shibayama,Yuzuru Kanakura,Naoki Hosen
    • Organizer
      第82回日本血液学会学術集会
  • [Presentation] Special AT-rich sequence binding protein 1 induces antigen presenting function in splenic B cells2020

    • Author(s)
      Takayuki Ozawa, Takafumi Yokota, Ritsuko Nakai, Yasuhiro Shingai, Tomoaki Ueda, Yukiko Doi, Hirohiko Shibayama, Naoki Hosen
    • Organizer
      第82回日本血液学会学術集会

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Published: 2021-12-27  

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