Role of CALR mutation in the pathogenesis of myeloproliferative neoplasms and rogue hematopoietic signaling

• Project/Area Number: 16K09852
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Hematology
• Research Institution: University of Miyazaki
• Principal Investigator: Shide Kotaro 宮崎大学, 医学部, 助教 (20468028)
• Research Collaborator: KAMEDA Takuro
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 骨髄線維症 / 骨髄増殖性腫瘍 / マウスモデル / 原発性骨髄線維症 / 分子標的療法 / Calreticulin / 癌 / シグナル伝達

Outline of Final Research Achievements

We analyzed Calreticulin (CALR), which is frequently mutated in essential thrombocythemia and primary myelofibrosis. The role of CALR in normal hematopoiesis and the mechanism by which mutant CALR causes MPN were elucidated in in vitro experiments and in in vivo experiments by creating multiple mouse models. We found that CALR mutation conferred cell proliferation and cytokine independence and caused mice to develop essential thrombocythemia. However, at the same time, it also became clear that they have a negative effect on hematopoietic stem cells, suggesting that maintenance of mutated hematopoietic stem cells and development of disease require elements other than CALR mutation.

Academic Significance and Societal Importance of the Research Achievements

原発性骨髄線維症の予後は5年生存率40%と不良であり、新規治療法へのアンメットメディカルニーズがある。CALR変異は原発性骨髄線維症の約3割の患者で認められることから治療標的として注目されている。本研究で見出したCALR変異による疾患発症の仕組みは、原発性骨髄線維症を始め、同じくCALR変異を有する本態性血小板血症の克服、治癒を目指した治療法開発につながる成果という意味で極めて意義が大きい。

Research Products

• [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] Mice with Calr mutations homologous to human CALR mutations only exhibit mild thrombocytosis (2019)
  • Author(s): Shide K、Kameda T、Kamiunten A、Oji A、Ozono Y、Sekine M、Honda A、Kitanaka A、Akizuki K、Tahira Y、Nakamura K、Hidaka T、Kubuki Y、Abe H、Miike T、Iwakiri H、Tahara Y、Sueta M、Hasuike S、Yamamoto S、Nagata K、Ikawa M、Shimoda K
  • Journal Title: Blood Cancer Journal Volume: 9 Issue: 4 Pages: 42-42
  • DOI: 10.1038/s41408-019-0202-z
  • NAID: 120006957457
  • 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] Early/prefibrotic primary myelofibrosis in patients who were initially diagnosed with essential thrombocythemia (2018)
  • Author(s): Kamiunten A, Shide K, Kameda T, Ito M, Sekine M, Kubuki Y, Hidaka T, Akizuki K, Tahira Y, Toyama T, Kawano N, Marutsuka K, Maeda K, Takeuchi M, Kawano H, Sato S, Ishizaki J, Shimoda H, Yamashita K, Matsuoka H, Shimoda K:
  • Journal Title: International Journal of Hematology Volume: 108 Issue: 4 Pages: 411-415
  • DOI: 10.1007/s12185-018-2495-2
  • Peer Reviewed / Open Access
• [Journal Article] Pathogenesis of myeloproliferative neoplasms: insights from mouse models (2018)
  • Author(s): 幣 光太郎
  • Journal Title: Rinsho Ketsueki Volume: 59 Issue: 10 Pages: 2075-2083
  • DOI: 10.11406/rinketsu.59.2075
  • NAID: 130007496566
  • ISSN: 0485-1439, 1882-0824
  • Peer Reviewed
• [Journal Article] Myeloproliferative neoplasms: recent progresses in therapy (2018)
  • Author(s): 上運天 綾子、幣 光太郎、下田 和哉
  • Journal Title: Rinsho Ketsueki Volume: 59 Issue: 6 Pages: 741-746
  • DOI: 10.11406/rinketsu.59.741
  • NAID: 130007393847
  • ISSN: 0485-1439, 1882-0824
  • Peer Reviewed
• [Journal Article] 原発性および二次性骨髄線維症の本邦における特徴 (特集 造血障害とその類縁疾患 : 最近の展開) (2018)
  • Author(s): 幣 光太郎、下田 和哉
  • Journal Title: 血液内科 Volume: 76 Pages: 59-65
  • NAID: 40021456609
• [Journal Article] 骨髄線維症に対する新たなJAK阻害薬momelotinibの臨床効果(SIMPLIFY-1/2試験) (2018)
  • Author(s): 幣 光太郎
  • Journal Title: 血液内科 Volume: 77 Pages: 131-136
  • NAID: 40021639666
• [Journal Article] 骨髄増殖性腫瘍の病態 (2018)
  • Author(s): 幣光太郎、亀田拓郎、下田和哉
  • Journal Title: サイトメトリーリサーチ Volume: 28 Pages: 7-12
  • NAID: 130007596141
• [Journal Article] Thrombohemorrhagic events, disease progression, and survival in polycythemia vera and essential thrombocythemia: a retrospective survey in Miyazaki prefecture, Japan (2018)
  • Author(s): Kamiunten A, Shide K, Kameda T, Sekine M, Kubuki Y, Ito M, Toyama T, Kawano N, Marutsuka K, Maeda K, Takeuchi M, Kawano H, Sato S, Ishizaki J, Akizuki K, Tahira Y, Shimoda H, Hidaka T, Yamashita K, Matsuoka H, Shimoda K.
  • Journal Title: Int J Hematol Volume: 印刷中 Issue: 6 Pages: 681-688
  • DOI: 10.1007/s12185-018-2428-0
  • Peer Reviewed / Open Access
• [Journal Article] Hmga2 collaborates with JAK2 V617F in the development of myeloproliferative neoplasms (2017)
  • Author(s): Ueda K, Ikeda K, Ikezoe T, Harada-Shirado K, Ogawa K, Hashimoto Y, Sano T, Ohkawara H, Kimura S, Shichishima-Nakamura A, Nakamura Y, Shikama Y, Mori T, Mason PJ, Bessler M, Morishita S, Komatsu N, Shide K, Shimoda K, Koide S, Aoyama K, Oshima M, Iwama Aand Takeishi Y.
  • Journal Title: Blood Advances Volume: 1 Issue: 15 Pages: 1001-1015
  • DOI: 10.1182/bloodadvances.2017004457
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Loss of Tyrosine Kinase 2 Does Not Affect the Severity of Jak2V617F-induced Murine Myeloproliferative Neoplasm (2017)
  • Author(s): Yamaji T, Shide K, Kameda T, Sekine M, Kamiunten A, Hidaka T, Kubuki Y, Shimoda H, Abe H, Miike T, Iwakiri H, Tahara Y, Sueta M, Yamamoto S, Hasuike S,Nagata K and Shimoda K Loss of Tyrosine Kinase 2 Does Not Affect the Severity of Jak2V617F-induced Murine Myeloproliferative Neoplasm
  • Journal Title: Anticancer Res Volume: 37 Issue: 7 Pages: 3841-3847
  • DOI: 10.21873/anticanres.11763
  • Peer Reviewed
• [Journal Article] CALR変異による本態性血小板血症モデル (2017)
  • Author(s): 幣 光太郎
  • Journal Title: 血液内科 Volume: 75 Pages: 358-363
  • NAID: 40021332151
• [Journal Article] Mutant calreticulin causes essential thrombocythemia (2017)
  • Author(s): Kazuya Shimoda, Kotaro Shide and Takuro Kameda
  • Journal Title: Oncotarget Volume: 8 Issue: 51 Pages: 88251-88252
  • DOI: 10.18632/oncotarget.21292
  • Peer Reviewed / Open Access
• [Journal Article] Differences in Hematological and Clinical Features Between Essential Thrombocythemia Cases With JAK2- or CALR-Mutations. (2017)
  • Author(s): Kubuki Y, Shide K, Kameda T, Yamaji T, Sekine M, Kamiunten A, Akizuki K, Shimoda H, Tahira Y, Nakamura K, Abe H, Miike T, Iwakiri H, Tahara Y, Sueta M, Hashimoto K, Yamamoto S, Hasuike S, Hidaka T, Nagata K, Kitanaka A, Shimoda K
  • Journal Title: Annals of laboratory medicine. Volume: 37 Issue: 2 Pages: 159-161
  • DOI: 10.3343/alm.2017.37.2.159
  • Peer Reviewed / Open Access
• [Journal Article] Calreticulin mutant mice develop essential thrombocythemia that is ameliorated by the JAK inhibitor ruxolitinib (2016)
  • Author(s): Kotaro Shide, Takuro Kameda, Takumi Yamaji, Masaaki Sekine, Naoki Inada, Ayako Kamiunten, Keiichi Akizuki, Kenichi Nakamura, Akira Kitanaka, Arata Honda, Akira Sawaguchi, Hiroo Abe, Tadashi Miike, Hisayoshi Iwakiri, Yoshihiro Tahara, Mitsue Sueta, Satoru Hasuike, Shojiro Yamamoto, Kenji Nagata, and Kazuya Shimoda
  • Journal Title: Leukemia Volume: 31 Issue: 5 Pages: 1136-1144
  • DOI: 10.1038/leu.2016.308
  • Peer Reviewed / Open Access
• [Presentation] Haploinsufficiency of Calr Confers Hematopoietic Stem Cells (HSCs) with a Clonal Advantage over Wild-Type Cells, and, in Setting of Myeloproliferative Neoplasms, Compensates for the Functions of HSCs Impaired By the Calr Mutation (2018)
  • Author(s): Shide K, Kameda T, Kamiunten A, Sekine A, Ozono Y, Yokomizo T, Akizuki K, Tahira Y, Kubuki Y, Hidaka T, Sashida G, Shimoda K
  • Organizer: American Society of Hematology 60th Annual Meeting, Abstract no. 0097, San Diego CA, 米国,Dec 1. 2018 (口演)
  • Int'l Joint Research
• [Presentation] TET2 Mutation Associated with Organ Infiltrations in ATLL (2018)
  • Author(s): Kameda T, Shide K, Kamiunten A, Tahira Y, Akizuki K, Sekine M, Hidaka T, Kubuki Y, Hidaka M, Utsunomiya A, Ohshima T, Kataoka K, Ogawa S, Shimoda K
  • Organizer: American Society of Hematology 60th Annual Meeting, Abstract no. 1345, San Diego CA, 米国,Dec 1. 2018 (ポスター)
  • Int'l Joint Research
• [Presentation] Pathogenesis of myeloproliferative neoplasms: insights from mouse models (Educational Lecture 3-3F) (2018)
  • Author(s): Shide K.
  • Organizer: The 80th Annual Meeting of the Japanese society of Hematology, EL3-3F, Osaka, JAPAN, Oct 14.2018 (教育講演)
  • Invited
• [Presentation] CALR haploinsufficiency compensate HSC funtion of CALR-del52 transgenic mice (2018)
  • Author(s): Shide K, Kameda T, Kamiunten A, Sekine M, Akizuki K, Tahira Y, Hidaka T, Kubuki Y, Ikawa M, Shimoda K.
  • Organizer: The 80th Annual Meeting of the Japanese society of Hematology, OS1-13A-5, Osaka, JAPAN, Oct 12.2018 (口演)
• [Presentation] JAK2 T875N, a causative novel activating germline mutation for familial essential thrombocytosis (2018)
  • Author(s): Yoshimitsu M, Hachiman M, Shide K, Shimoda K, Ishitsuka K
  • Organizer: The 80th Annual Meeting of the Japanese society of Hematology, OS1-13B-1, Osaka, JAPAN, Oct 12.2018 (口演)
• [Presentation] Generation of Calr mutation knock-in mice using CRISPR/Cas9 system (2018)
  • Author(s): Shide K, Ikawa M and Shimoda K
  • Organizer: Japan Society for the Promotion of Science & National University of Singapore Joint 2nd symposium, NEW HORIZONS IN NORMAL AND CANCER STEM CELL RESEARCH
  • Int'l Joint Research
• [Presentation] Murine Calr mutants homologous to human CALR mutants activate STATs, but do not develop MPNs in vivo (2017)
  • Author(s): Shide K, Kameda T, Sekine M, Kamiunten A, Akizuki K, Tahira Y, Hidaka T, Kubuki Y, Honda A, Ikawa M and Shimoda K
  • Organizer: 第79回日本血液学会学術集会
  • Int'l Joint Research
• [Presentation] CALR del52 変異はMPLと協調してJAK-STAT経路を活性化し、マウスにETを発症させる (2017)
  • Author(s): 幣 光太郎
  • Organizer: 第54回日本臨床分子医学会学術集会
• [Presentation] Physiological Expression of Calr Mutant induces Increased Cell Growth and Cytokine Independency in Human Cell Lines Expressing Mpl, and Develops Essential Thrombocythemia in Mice (2016)
  • Author(s): Shide K, Kameda T, Sekine A, Kamiunten A, Akizuki K, Tahira Y, Hidaka T, Kubuki Y, Honda A, Sawaguchi A, Kitanaka A and Shimoda K.
  • Organizer: American Society of Hematology 58th Annual Meeting
  • Place of Presentation: サンディエゴ(米国)
  • Int'l Joint Research
• [Presentation] HMGA2 Orchestrates the Tumorgenesis of Myeloproliferative Neoplasms (MPN) in Corporation with JAK2V617F (2016)
  • Author(s): Ueda K, Ikeda K, Ikezoe T, Ogawa K, Hashimoto Y, Harada-Shirado K, Ohkawara H, Komatsu N, Shide K, Shimoda K, Koide S, Oshima M, Iwama A and Takeishi Y.
  • Organizer: American Society of Hematology 58th Annual Meeting
  • Place of Presentation: サンディエゴ(米国)
  • Int'l Joint Research
• [Presentation] CALRdel52 mutation activates the JAK-STAT pathway in coordination with MPL, and causes ET in mice (2016)
  • Author(s): Shide K, Kameda T, Sekine M, Kamiunten A, Akizuki K, Tahira Y, Shimoda H, Hidaka T, Kubuki Y, Honda A, Kitanaka A, Shimoda K.
  • Organizer: 第78回日本血液学会
  • Place of Presentation: 横浜
  • Int'l Joint Research
• [Presentation] The role of oncogene HMGA2 in the pathogenesis of myeloproliferative neoplasms (MPN) (2016)
  • Author(s): Ueda K, Ikeda K, Ogawa K, Hashimoto Y, Morishita S, Komatsu N, Shide K, Shimoda K, Iwama A, Takeishi A.
  • Organizer: 第78回日本血液学会
  • Place of Presentation: 横浜
  • Int'l Joint Research