Control of myelofibrosis targeting the amplification mechanism of CALR mutant stem cells and the functional motif of CALR mutant proteins

• Project/Area Number: 19K08819
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 54010:Hematology and medical oncology-related
• Research Institution: University of Miyazaki
• Principal Investigator: Shide Kotaro 宮崎大学, 医学部, 助教 (20468028)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 骨髄増殖性腫瘍 / 原発性骨髄線維症 / 造血幹細胞 / 分子標的療法 / Calreticulin

Outline of Research at the Start

骨髄線維症患者の5年生存率は40%と予後不良である。この疾患群のドライバー変異であるcalreticulin(CALR)の変異は造血幹細胞レベルで生じており、約3割の患者にみられる。本研究では、申請者が見出した同変異に関する最新の研究成果を基盤として、「CALR変異造血幹細胞のクローン拡大機序」、「CALR変異蛋白による増殖シグナル活性化機序」を解明する。更に、両機序を標的とした病態制御のproof of conceptを得て、骨髄線維症の革新的治療に道を開くことを目標とする。

Outline of Final Research Achievements

We created the first blood cell-specific Calreticulin (Calr) KO mouse that had not yet been developed, and clarified the role of Calr in hematopoiesis. We further showed that haploinsufficiency of the Calr gene enhances the competitive repopulation capacity of hematopoietic stem cells (HSCs), and also enhances the clonal expansion of HSCs with CALR mutations in myeloproliferative patients.
Regarding the mechanism of cell proliferation by CALR mutant protein, we found a sequence essential for tumorigenicity in vitro and in vivo in the amino acid sequence specific to the mutant protein. We plan to begin preclinical studies targeting these amino acids.

Academic Significance and Societal Importance of the Research Achievements

原発性骨髄線維症(primary myelofibrosis:PMF) 、および本態性血小板血症(essential thrombocythemia:ET)に続発する二次性骨髄線維症(post-ET MF)患者の5年生存率は40%と予後不良であり、PMF, post-ET MFには臨床的なunmet needsがある。この疾患群のドライバー変異であるcalreticulin(CALR)の変異は造血幹細胞レベルで生じており、約3割の患者にみられる。本研究の成果は、CALR変異が疾患を引き起こす仕組みを明らかにしたものであり、これらの患者群を対象とした新規治療開発の基盤となるものである。

Research Products

• [Journal Article] Immunohistopathological Analysis of Extramedullary Hematopoiesis and Angiogenesis of Spleen in a Case of Primary Myelofibrosis with Huge Splenomegaly (2022)
  • Author(s): Kawano N, Saito N, Yoshida S, Kitanaka A, Shide K, Marutsuka K, Ohshima K, Shimoda K
  • Journal Title: The Tohoku Journal of Experimental Medicine Volume: 256 Issue: 2 Pages: 119-125
  • DOI: 10.1620/tjem.256.119
  • NAID: 130008160463
  • ISSN: 0040-8727, 1349-3329
  • Peer Reviewed / Open Access
• [Journal Article] Clonal hematopoiesis with JAK2V617F promotes pulmonary hypertension with ALK1 upregulation in lung neutrophils (2021)
  • Author(s): Kimishima Yusuke、Misaka Tomofumi、Yokokawa Tetsuro、Wada Kento、Ueda Koki、Sugimoto Koichi、Minakawa Keiji、Nakazato Kazuhiko、Ishida Takafumi、Oshima Motohiko、Koide Shuhei、Shide Kotaro、Shimoda Kazuya、Iwama Atsushi、Ikeda Kazuhiko、Takeishi Yasuchika
  • Journal Title: Nature Communications Volume: 12 Issue: 1 Pages: 6177-6177
  • DOI: 10.1038/s41467-021-26435-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Fibrocytes in primary myelofibrosis (2021)
  • Author(s): Kazuya Shimoda, Yoshinori Ozono, Kotaro Shide
  • Journal Title: Oncotarget Volume: 12 Issue: 20 Pages: 2101-2103
  • DOI: 10.18632/oncotarget.27971
  • Peer Reviewed / Open Access
• [Journal Article] Calreticulin mutations in myeloproliferative neoplasms (2021)
  • Author(s): Kotaro Shide
  • Journal Title: Int Rev Cell Mol Biol Volume: 365 Pages: 179-226
  • DOI: 10.1016/bs.ircmb.2021.05.005
  • ISBN: 9780323899390
  • Peer Reviewed
• [Journal Article] Pathophysiology of myeloproliferative neoplasms revealed from mouse models with driver mutations. (2020)
  • Author(s): 幣 光太郎
  • Journal Title: Cytometry Research Volume: 30 Issue: 2 Pages: 15-20
  • DOI: 10.18947/cytometryresearch.30.2_15
  • NAID: 130007952539
  • ISSN: 0916-6920, 2424-0664
  • Year and Date: 2020-12-11
  • Peer Reviewed / Open Access
• [Journal Article] Calreticulin haploinsufficiency augments stem cell activity and is required for onset of myeloproliferative neoplasms (2020)
  • Author(s): Shide Kotaro、Kameda Takuro、Kamiunten Ayako、Ozono Yoshinori、Tahira Yuki、Yokomizo-Nakano Takako、Kubota Sho、Ono Masaya、Ikeda Kazuhiko、Sekine Masaaki、Akizuki Keiichi、Nakamura Kenichi、Hidaka Tomonori、Kubuki Yoko、Iwakiri Hisayoshi、Hasuike Satoru、Nagata Kenji、Sashida Goro、Shimoda Kazuya
  • Journal Title: Blood Volume: - Pages: 106-118
  • DOI: 10.1182/blood.2019003358
  • Peer Reviewed / Open Access
• [Journal Article] Neoplastic fibrocytes play an essential role in bone marrow fibrosis in Jak2V617F-induced primary myelofibrosis mice (2020)
  • Author(s): Ozono Yoshinori、Shide Kotaro、Kameda Takuro、Kamiunten Ayako、Tahira Yuki、Sekine Masaaki、Akizuki Keiichi、Nakamura Kenichi、Iwakiri Hisayoshi、Sueta Mitsue、Hidaka Tomonori、Kubuki Yoko、Yamamoto Shojiro、Hasuike Satoru、Sawaguchi Akira、Nagata Kenji、Shimoda Kazuya
  • Journal Title: Leukemia Volume: - Issue: 2 Pages: 454-467
  • DOI: 10.1038/s41375-020-0880-3
  • Peer Reviewed / Open Access
• [Journal Article] JAK2-negative acute monocytic leukemia with TET2 mutation in essential thrombocythemia with JAK2 mutation with literature review (2020)
  • Author(s): Ogasawara Toshie、Kawauchi Kiyotaka、Ono Takuya、Marshall Shoko、Shide Kotaro、Shimoda Kazuya、Mori Naoki、Sakura Hiroshi
  • Journal Title: Leukemia Research Reports Volume: 13 Pages: 100194-100194
  • DOI: 10.1016/j.lrr.2019.100194
  • Peer Reviewed / Open Access
• [Journal Article] The role of driver mutations in myeloproliferative neoplasms: insights from mouse models (2020)
  • Author(s): Kotaro Shide
  • Journal Title: International Journal of Hematology Volume: 111 Issue: 2 Pages: 206-216
  • DOI: 10.1007/s12185-019-02803-x
  • Peer Reviewed / Open Access
• [Journal Article] TP53 and PTEN Mutations Were Shared in Concurrent Germ Cell Tumor and Acute Megakaryoblastic Leukemia (2020)
  • Author(s): Keiichi Akizuki, Masaaki Sekine, Yasunori Kogure, Takuro Kameda, Kotaro Shide, Junji Koya, Ayako Kamiunten, Yoko Kubuki, Yuki Tahira, Tomonori Hidaka, Takumi Kiwaki, Hiroyuki Tanaka, Yuichiro Sato, Hiroaki Kataoka, Keisuke Kataoka & Kazuya Shimoda
  • Journal Title: BMC Cancer Volume: 20 Issue: 1 Pages: 5-5
  • DOI: 10.1186/s12885-019-6497-0
  • Peer Reviewed / Open Access
• [Journal Article] Essential thrombocytosis attributed to JAK2-T875N germline mutation (2019)
  • Author(s): Makoto Yoshimitsu, Miho Hachiman, Yuichiro Uchida, Naosuke Arima, Akihiko Arai, Yuhei Kamada, Kotaro Shide, Masafumi Ito, Kazuya Shimoda, Kenji Ishitsuka
  • Journal Title: International Journal of Hematology Volume: 110 Issue: 5 Pages: 584-590
  • DOI: 10.1007/s12185-019-02725-8
  • Peer Reviewed / Open Access
• [Journal Article] Monocyte-derived fibrocytes elimination had little contribution on liver fibrosis (2019)
  • Author(s): Ozono Yoshinori、Shide Kotaro、Toyoshima Fumiyo、Takaishi Yuuka、Tsuchimochi Mai、Kamiunten Ayako、Kameda Takuro、Nakamura Kenichi、Miike Tadashi、Kusumoto Kazunori、Iwakiri Hisayoshi、Hasuike Satoru、Nagata Kenji、Sawaguchi Akira、Shimoda Kazuya
  • Journal Title: Hepatobiliary & Pancreatic Diseases International Volume: Epub ahead of print Issue: 4 Pages: 348-353
  • DOI: 10.1016/j.hbpd.2019.02.002
  • Peer Reviewed / Open Access
• [Journal Article] Vitamin D receptor?mediated skewed differentiation of macrophages initiates myelofibrosis and subsequent osteosclerosis (2019)
  • Author(s): Wakahashi Kanako、Minagawa Kentaro、Kawano Yuko、Kawano Hiroki、Suzuki Tomohide、Ishii Shinichi、Sada Akiko、Asada Noboru、Sato Mari、Kato Shigeaki、Shide Kotaro、Shimoda Kazuya、Matsui Toshimitsu、Katayama Yoshio
  • Journal Title: Blood Volume: 133 Issue: 15 Pages: 1619-1629
  • DOI: 10.1182/blood-2018-09-876615
  • NAID: 120007002632
  • Peer Reviewed / Open Access
• [Journal Article] CALR変異によるMPN発症機構 (2019)
  • Author(s): 幣光太郎
  • Journal Title: 血液内科 Volume: 78 Pages: 678-674
  • NAID: 40021930128
• [Journal Article] 骨髄増殖性腫瘍におけるJAK阻害薬の使用基準 (2019)
  • Author(s): 3.幣光太郎, 下田和哉
  • Journal Title: 血液内科 Volume: 78 Pages: 870-876
  • NAID: 40021948459
• [Journal Article] マウスモデルによるMPNの病態解析 (2019)
  • Author(s): 幣光太郎
  • Journal Title: 血液内科 Volume: 79 Pages: 542-549
• [Journal Article] 骨髄増殖性腫瘍の病期進展メカニズム (2019)
  • Author(s): 幣光太郎, 下田和哉
  • Journal Title: 血液内科 Volume: 79 Pages: 697-701
• [Presentation] Role of driver mutations revealed from MPN mouse models. (2021)
  • Author(s): Kotaro Shide
  • Organizer: The 12th JSH International Symposium 2021 in Kamaura, AS II-3, Kamakura JAPAN (WEB併用)
  • Int'l Joint Research / Invited
• [Presentation] Hematopoietic cells with MPN driver mutations promote pulmonary hypertension (2021)
  • Author(s): Kazuhiko Ikeda，Tomofumi Misaka，Yusuke Kimishima， Tetsuro Yokokawa，Keiji Minakawa，Koki Ueda，Motohiko Oshima， Shuhei Koide，Kotaro Shide，Kazuya Shimoda，Osamu Nakajima， Atsushi Iwama，Yasuchika Takeishi
  • Organizer: The 83th Annual Meeting of the Japanese society of Hematology, OS2-7A-14, Sendai JAPAN, Sep 24.2021 (口演)
• [Presentation] Loss of EZH2 does not affect the severity of murine myeloproliferative neoplasms with CALR mutation (2021)
  • Author(s): Ayako Kamiunten, Yuki Tahira, Keiichi Akizuki, Masaaki Sekine, Takuro Kameda, Kotaro Shide, Tomonori Hidaka, Yoko Kubuki, Kazuya Shimoda
  • Organizer: The 83th Annual Meeting of the Japanese society of Hematology, OS2-7A-3, Sendai JAPAN, Sep 24.2021 (口演)
• [Presentation] RNA splicing machinery gene mutations in patients with myeloproliferative neoplasms (MPNs) in Japan (2021)
  • Author(s): Kotaro Shide, Saki Ogawa, Ayako Kamiunten et al.
  • Organizer: The 83th Annual Meeting of the Japanese society of Hematology, OS2-7B-4, Sendai JAPAN, Sep 24.2021 (口演)
• [Presentation] Renal impairment associated with myeloproliferative neoplasm (MPN) (2021)
  • Author(s): Yuki Tahira, Kotaro Shide, Ayako Kamiunten, Takuro Kameda, Keiichi Akizuki, Masaaki Sekine, Tomonori Hidaka, Yoko Kubuki, Kazuya Shimoda
  • Organizer: The 83th Annual Meeting of the Japanese society of Hematology, PS-11-6, Sendai JAPAN, Sep 23.2021 (ポスター)
• [Presentation] はじめての血液学2 、血球増加症へのアプローチ (2020)
  • Author(s): Kotaro Shide
  • Organizer: The 82th Annual Meeting of the Japanese society of Hematology, SN-2, Kyoto JAPAN (WEB開催), Oct 10.2020 (教育講演)
  • Invited
• [Presentation] Mechanism of enhanced IL1α production induced by JAK2V617F (2020)
  • Author(s): Kotaro Shide，Ayako Kamiunten，Yoshinori Ozono，Takuro Kameda，Yuki Tahira，Keiichi Akizuki，Masaaki Sekine，Tomonori Hidaka，Yoko Kubuki，Kazuya Shimoda
  • Organizer: The 82th Annual Meeting of the Japanese society of Hematology, OS-25-5, Kyoto JAPAN (WEB開催), Oct 10.2020 (口演)
• [Presentation] Myeloproliferative neoplasm (MPN) nephropathy in a Jak2V617F induced MPN mouse model (2020)
  • Author(s): Yuki Tahira，Kotaro Shide，Yoshinori Ozono，Ayako Kamiunten，Takuro Kameda，Keiichi Akizuki，Masaaki Sekine，Tomonori Hidaka，Yoko Kubuki，Kazuya Shimoda
  • Organizer: The 82th Annual Meeting of the Japanese society of Hematology, PS-12-6, Kyoto JAPAN (WEB開催), Oct 10.2020 (ポスター)
• [Presentation] SRSF2, U2AF1, and IDH mutations in patients with myeloproliferative neoplasms (MPNs) in Japan (2020)
  • Author(s): Ogawa S, Shide K, Kamiunten A, Kubuki Y, Takeuchi M, Matsue K, Shimomura T, Suzushima H, Kawano N, Yamashita K, Yasumi M, Karasuno T, Imataki O, Kadowaki N, Yonezawa A, Otsuka E, Hidaka T, Kameda T, Akizuki K, Sekine M, Tahira Y, Shimoda K
  • Organizer: The 82th Annual Meeting of the Japanese society of Hematology, OS-27-4, Kyoto JAPAN (WEB開催), Oct 10.2020 (口演)
• [Presentation] A case of 5q deletion syndrome with CALR mutation (2020)
  • Author(s): Shiho Fujiwara，Kotaro Shide，Kazuya Shimoda，Nobuyuki Arima，Yukiko Kimura，Yuko Watanabe，Taizo Shimomura，Hitoshi Suzushima
  • Organizer: The 82th Annual Meeting of the Japanese society of Hematology, PS-15-10, Kyoto JAPAN (WEB開催), Oct 10.2020 (ポスター)
• [Presentation] Clonal Hematopoiesis With JAK2V617F Promotes Pulmonary Hypertension Through ALK1 (2020)
  • Author(s): Yusuke Kirishima, Tomofumi Misaka, Tetsuro Yokokawa, Kento Wada, Koki Ueda, Koichi Sugimoto, Keiji Minakawa, Kazuhiko Nakazato, Takafumi Ishida, Motohiko Oshima, Shuhei Koide, Kotaro Shide, Kazuya Shimoda, Atsushi Iwama, Kazuhiko Ikeda, Yasuchika Takeishi
  • Organizer: American Heart Association Scientific Session 2020, abstract 12873
  • Int'l Joint Research
• [Presentation] The Role of Calreticulin in Normal Hematopoiesis and Neoplastic Hematopoiesis of Myeloproliferative Neoplasms (2019)
  • Author(s): Kotaro Shide, Takuro Kameda, Ayako Kamiunten, Yoshinori Ozono, Yuki Tahira, Masaaki Sekine, Masaya Ono, Takako Yokomizo, Sho Kubota, Keiichi Akizuki, Yoko Kubuki, Tomonori Hidaka, Goro Sashida, Kazuya Shimoda
  • Organizer: American Society of Hematology the 61st Annual Meeting and Exposition
  • Int'l Joint Research
• [Presentation] Depletion of Neoplastic CD11b Positive Cells in Jak2V617F Mutant Mice Reduced Fibrocytes in Bone Marrow and Improved Myelofibrosis (2019)
  • Author(s): Yoshinori Ozono, Kotaro Shide, Takuro Kameda, Ayako Kamiunten, Yuki Tahira, Masaaki Sekine, Keiichi Akizuki, Kenichi Nakamura, Hisayoshi Iwakiri, Tomonori Hidaka, Yoko Kubuki, Satoru Hasuike, Fumiyo Toyoshima, Akira Sawaguchi, Kenji Nagata, Kazuya Shimoda
  • Organizer: American Society of Hematology the 61st Annual Meeting and Exposition
  • Int'l Joint Research
• [Presentation] Calreticulin不全が造血系に与える影響 (2019)
  • Author(s): 幣光太郎、亀田拓郎、上運天綾子、大園芳徳、尾野雅哉、横溝貴子、久保田翔、田平優貴、秋月渓一、関根雅明、久冨木庸子、日高智徳、指田吾郎、伊川正人、下田和哉
  • Organizer: The 81th Annual Meeting of the Japanese society of Hematology
• [Presentation] マウスモデルを用いた骨髄増殖性腫瘍発症機構の解析 (2019)
  • Author(s): 幣 光太郎
  • Organizer: 第29回日本サイトメトリー学会学術集会
  • Invited