Establishment of therapeutic concept targeting homomultimerized mutant CALR

• Project/Area Number: 19K08848
• 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: Juntendo University
• Principal Investigator: ARAKI MARITO 順天堂大学, 大学院医学研究科, 客員教授 (80613843)
• Co-Investigator(Kenkyū-buntansha): 小松 則夫 順天堂大学, 医学部, 特任教授 (50186798)
• 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: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 骨髄増殖性腫瘍 / calreticulin / トロンボポエチン / JAK2 / 分子シャペロン / 受容体 / ２量体 / カルレティキュリン / MPL

Outline of Research at the Start

本研究では、変異型CALR上においてホモ多量体化を担っているドメインを同定し、質量分析やX線結晶構造解析により得られたホモ多量体化ドメインにおいて、多量体化を制御している翻訳後修飾を明らかにする。続いて、このような翻訳後修飾による変異型CALRの多量体化を、RNA干渉法あるいは阻害剤により抑制することで、変異型CALRによる細胞の腫瘍性増殖が抑制できることを示す。最後に、患者細胞や動物モデルにおいて、変異型CALRの多量体化を標的として治療概念を証明する。

Outline of Final Research Achievements

We studied an oncogenic domain that is generated by the frameshift mutation on the carboxyl-terminal of mutant CALR, a causal factor of Philadelphia-negative myelproliferative neoplasms. As a result, we found a new intramolecular interaction mediated by this domain that does not exist in wild-type CALR, identified amino acid residues involved in the interaction, and clarified the significance of the interaction in the oncogenic capacity of mutnat CALR.

Academic Significance and Societal Importance of the Research Achievements

本研究により、変異型CALRに腫瘍原性を付与するドメインの有する機能が明らになったことから、CALR遺伝子変異を有するMPN患者に対する有効な治療薬の開発に資する情報が取得できたことは大変意義深いと考えられる。さらに、変異型分子シャペロンによる受容体活性化という、生物学的にユニークな現象の分子メカニズムの一端が明らかにされたことから、この知見に基づいて、これまで未知となっている生命現象の解明が期待される。

Research Products

• [Journal Article] MPL overexpression induces a high level of mutant-CALR/MPL complex: a novel mechanism of ruxolitinib resistance in myeloproliferative neoplasms with CALR mutations (2021)
  • Author(s): Yasuda Shunichiro、Aoyama Satoru、Yoshimoto Ryoto、Li Huixin、Watanabe Daisuke、Akiyama Hiroki、Yamamoto Kouhei、Fujiwara Takeo、Najima Yuho、Doki Noriko、Sakaida Emiko、Edahiro Yoko、Imai Misa、Araki Marito、Komatsu Norio、Miura Osamu、Kawamata Norihiko
  • Journal Title: International Journal of Hematology Volume: 114 Issue: 4 Pages: 424-440
  • DOI: 10.1007/s12185-021-03180-0
  • Peer Reviewed
• [Journal Article] Cell-autonomous megakaryopoiesis associated with polyclonal hematopoiesis in triple-negative essential thrombocythemia (2021)
  • Author(s): Inano Tadaaki、Araki Marito、Morishita Soji、Imai Misa、Kihara Yoshihiko、Okuda Maho、Yang Yinjie、Ito Masafumi、Osaga Satoshi、Mano Hiroyuki、Edahiro Yoko、Ochiai Tomonori、Misawa Kyohei、Fukuda Yasutaka、Ando Jun、Komatsu Norio
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 17702-17702
  • DOI: 10.1038/s41598-021-97106-9
  • Peer Reviewed / Open Access
• [Journal Article] MPNにおけるcalreticulin遺伝子変異の意義 (2021)
  • Author(s): 増渕菜弥、荒木真理人、小松則夫
  • Journal Title: 血液内科 Volume: 82 Pages: 339-346
• [Journal Article] Mechanism underlying the development of myeloproliferative neoplasms through mutant calreticulin (2020)
  • Author(s): Edahiro Yoko、Araki Marito、Komatsu Norio
  • Journal Title: Cancer Science Volume: 111 Issue: 8 Pages: 2682-2688
  • DOI: 10.1111/cas.14503
  • Peer Reviewed / Open Access
• [Journal Article] CREB3L1 overexpression as a potential diagnostic marker of Philadelphia chromosome?negative myeloproliferative neoplasms (2020)
  • Author(s): Morishita Soji、Yasuda Hajime、Yamawaki Saya、Kawaji Hideya、Itoh Masayoshi、Edahiro Yoko、Imai Misa、Kogo Yasushi、Tsuneda Satoshi、Ohsaka Akimichi、Hayashizaki Yoshihide、Ito Masafumi、Araki Marito、Komatsu Norio
  • Journal Title: Cancer Science Volume: 112 Issue: 2 Pages: 884-892
  • DOI: 10.1111/cas.14763
  • Peer Reviewed / Open Access
• [Journal Article] Thrombocytosis caused by abnormal molecular chaperone (2020)
  • Author(s): ARAKI Marito
  • Journal Title: Japanese Journal of Thrombosis and Hemostasis Volume: 31 Issue: 5 Pages: 491-497
  • DOI: 10.2491/jjsth.31.491
  • NAID: 130007928076
  • ISSN: 0915-7441, 1880-8808
  • Peer Reviewed
• [Journal Article] 変異型calreticulinによる骨髄増殖性腫瘍発症の分子基盤 (2020)
  • Author(s): 荒木 真理人
  • Journal Title: 臨床血液 Volume: 61 Pages: 937-944
  • NAID: 130007897793
  • Peer Reviewed
• [Journal Article] Mutant calreticulin interacts with MPL in the secretion pathway for activation on the cell surface (2019)
  • Author(s): Masubuchi Nami、Araki Marito、Yang Yinjie、Hayashi Erina、Imai Misa、Edahiro Yoko、Hironaka Yumi、Mizukami Yoshihisa、Kihara Yoshihiko、Takei Hiraku、Nudejima Mai、Koike Masato、Ohsaka Akimichi、Komatsu Norio
  • Journal Title: Leukemia Volume: 34 Issue: 2 Pages: 499-509
  • DOI: 10.1038/s41375-019-0564-z
  • Peer Reviewed
• [Journal Article] The role of calreticulin mutations in myeloproliferative neoplasms (2019)
  • Author(s): Araki Marito、Komatsu Norio
  • Journal Title: International Journal of Hematology Volume: 111 Issue: 2 Pages: 200-205
  • DOI: 10.1007/s12185-019-02800-0
  • Peer Reviewed / Open Access
• [Journal Article] JAK2 exon 12 mutation in myelodysplastic/myeloproliferative neoplasm with ring sideroblasts and thrombocytosis: Not an exclusive mutation to polycythaemia vera (2019)
  • Author(s): Inano, T. Araki, M. Morishita, S. Imai, M. Yasuda, H. Nitta, H. Ito, M. Edahiro, Y. Ochiai, T. Misawa, K. Fukuda, Y. Ohsaka, A. Komatsu, N.
  • Journal Title: British Journal of Haematology Volume: 187 Issue: 1
  • DOI: 10.1111/bjh.16146
  • Peer Reviewed
• [Journal Article] Evidence for prevention of renal dysfunction associated with primary myelofibrosis by cytoreductive therapy (2019)
  • Author(s): Fukuda Yasutaka、Araki Marito、Yamamoto Kouji、Morishita Soji、Inano Tadaaki、Misawa Kyohei、Ochiai Tomonori、Edahiro Yoko、Imai Misa、Yasuda Hajime、Gotoh Akihiko、Ohsaka Akimichi、Komatsu Norio
  • Journal Title: Haematologica Volume: 印刷中 Issue: 11 Pages: e506-e509
  • DOI: 10.3324/haematol.2018.208876
  • Peer Reviewed / Open Access
• [Journal Article] Clinical and molecular features of patients with prefibrotic primary myelofibrosis previously diagnosed as having essential thrombocythemia in Japan (2019)
  • Author(s): Edahiro Yoko、Araki Marito、Inano Tadaaki、Ito Masafumi、Morishita Soji、Misawa Kyohei、Fukuda Yasutaka、Imai Misa、Ohsaka Akimichi、Komatsu Norio
  • Journal Title: European Journal of Haematology Volume: 00 Issue: 6 Pages: 1-5
  • DOI: 10.1111/ejh.13236
  • Peer Reviewed
• [Journal Article] MPNのゲノム解析研究の進歩 (2019)
  • Author(s): 奥田真帆、荒木真理人、小松則夫
  • Journal Title: 血液内科 Volume: 79 Pages: 556-564
• [Journal Article] 骨髄増殖性腫瘍の遺伝子異常 (2019)
  • Author(s): 荒木真理人、森下総司、小松則夫
  • Journal Title: 臨床検査 Volume: 63 Pages: 790-796
• [Presentation] On the mechanism of sugar-chain mediated activation of thrombopoietin receptor (MPL) (2021)
  • Author(s): Hiromi Watari, Hiromu Kageyama, Hiroya Nakajima, Kako Onodera, PJ Foces, Nami Masubuchi, Takashi Matsui, Yoshio Kodera, Tomohisa Ogawa, Makoto Hirayama, Kanji Hori, Takeshi Yokoyama, Douglas Freymann, Norio Komatsu, Marito Araki, Yoshikazu Tanaka, Ryuichi Sakai
  • Organizer: 第83回日本血液学会学術集会
• [Presentation] Therapeutic Potential of an Antibody Targeting the Cleaved Form of Mutant Calreticulin in Myeloproliferative Neoplasms (2021)
  • Author(s): Marito Araki
  • Organizer: MPN Asia 2021 - the International Symposium on Myeloproliferative Neoplasms
  • Int'l Joint Research / Invited
• [Presentation] Distributions of typical non-driver mutations in essential thrombocythemia and polycythemia vera (2021)
  • Author(s): Soji Morishita, Yoko Edahiro, Yoshinori Hashimoto, Shuichi Shirane, Tomonori Ochiai, Kyohei Misawa, Tadaaki Inano, Yasutaka Fukuda, Yosuke Mori, Chiho Furuya, Marito Araki, Jun Ando, Norio Komatsu
  • Organizer: 第83回日本血液学会学術集会
• [Presentation] 海洋天然物由来の新規生理活性タンパク質の構造機能解析 (2021)
  • Author(s): 田中良和, 影山大夢, 辺ひろみ, 増渕菜弥, 中島寛也, 小野寺かこ, 松井崇, 小寺義男, 小川智久, 横山武司, 小松則夫, 荒木真理人, 酒井隆一
  • Organizer: 第94回日本生化学会大会
  • Invited
• [Presentation] Quantitative measurement of mutant calreticulin in specimens of myeloproliferative neoplasm patients (2021)
  • Author(s): Satoko Ogata, Marito Araki, Yoshihiko Kihara, Maho Okuda, Soji Morishita, Misa Imai, Nami Masubuchi, Yousuke Mori, Yinjie Yang, Syunpei Yoshikawa, Tomonori Ochiai, Shuichi Shirane, Yoko Edahiro, Jun Ando, Norio Komatsu
  • Organizer: 第83回日本血液学会学術集会
• [Presentation] Therapeutic potential of an antibody targeting the cleaved form of mutant calreticulin in myeloproliferative neoplasms (2020)
  • Author(s): Kihara Y, Araki M, Imai M, Mori Y, Horino M, Ogata T, Yoshikawa S, Taguchi T, Masubuchi N, Mabuchi Y, Yang Y, Fukuda Y, Morishita S, Suzuki T, Dohmae N, Shimonaka M, Akazawa C, Ohsaka A, Komatsu N.
  • Organizer: 62nd Annual Meeting & Exposition of American Society of Hematology
  • Int'l Joint Research
• [Presentation] Concomitant occurrence of polyclonal hematopoiesis and cell-autonomous megakaryopoiesis in triple-negative essential thrombocythemia (2020)
  • Author(s): Inano T, Araki M, Morishita S, Imai M, Kihara Y, Okuda M, Ito M, Osaga S, Yang Y, Edahiro Y, Ochiai T, Misawa K, Fukuda Y, Ohsaka A, Komatsu N.
  • Organizer: 62nd Annual Meeting & Exposition of American Society of Hematology
  • Int'l Joint Research
• [Presentation] CREB3L1 Overexpression Can Reliably Discriminate Ph-MPNs from Reactive Cases (2020)
  • Author(s): Morishita S, Yasuda H, Yamawaki S, Kawaji H, Itoh M, Edahiro Y, Imai M, Kogo Y, Tsuneda S, Akimichi Ohsaka, Hayashizaki Y, Araki M, Komatsu N.
  • Organizer: 62nd Annual Meeting & Exposition of American Society of Hematology
  • Int'l Joint Research
• [Presentation] MPL Overexpression Induces a High Level of Mutant-Calr/MPL Complex: A Novel Mechanism of Ruxolitinib Resistance in Myeloproliferative Neoplasms with Calr Mutations (2020)
  • Author(s): Yasuda S, Aoyama S, Yoshimoto R, Watanabe D, Akiyama H, Yamamoto K, Fujiwara T, Sakaida E, Edahiro Y, Imai M, Araki M, Komatsu N, Kawamata N.
  • Organizer: 62nd Annual Meeting & Exposition of American Society of Hematology
  • Int'l Joint Research
• [Presentation] Therapeutic potential of myeloproliferative neoplasms by antibody targeting mutant calreticulin (2020)
  • Author(s): Kihara Y, Araki M, Imai M, Fukuda Y, Mori Y, Taguchi T, Masubuchi N, Mabuchi Y, Yang Y, Mizukami Y, Morishita S, Akazawa C, Ohsaka A, Komatsu N.
  • Organizer: 第82回日本血液学会学術集会
• [Presentation] Analysis of behavior of erythropoietin receptor on the cell surface in JAK2 mutant cells (2020)
  • Author(s): Shioiri K, Araki M, Kitazawa S, Masubuchi N, Morishita S, Imai M, Ohsaka A, Komatsu N.
  • Organizer: 第82回日本血液学会学術集会
• [Presentation] CREB3L1 overexpression as a novel diagnostic marker of myeloproliferative neoplasms (2020)
  • Author(s): Morishita S, Yasuda H, Yamawaki S, Kawaji H, Itoh M, Edahiro Y, Imai M, Kogo Y, Tsuneda S, Ohsaka A, Hayashizaki Y, Araki M, Komatsu N.
  • Organizer: 第82回日本血液学会学術集会
• [Presentation] Targeting ER stress enhances inhibitory effect of ruxolitinib in mutated-CALR transfected cells (2020)
  • Author(s): Moriyama M, Akahane D, Moriya S, Imai M, Araki M, Komatsu N, Miyazawa K, Gotoh A.
  • Organizer: 第82回日本血液学会学術集会
• [Presentation] Overexpression of MPL causes ruxolitinib-resistance in MPN with CALR frame-shift mutations. (2020)
  • Author(s): Yasuda S, Aoyama S, Yoshimoto R, Watanabe D, Yamamoto K, Fujiwara T, Edahiro Y, Imai M, Araki M, Komatsu N, Miura O, Kawamata N.
  • Organizer: 第82回日本血液学会学術集会
• [Presentation] Targeting calreticulin neoantigen for the treatment of myeloproliferative neoplasms (2020)
  • Author(s): Araki M, Komatsu N.
  • Organizer: 第79回日本癌学会学術総会
• [Presentation] 典型的なドライバー遺伝子変異の見いだされない本態性血小板血症の特徴 (2020)
  • Author(s): 荒木真理人, 稲野資明, 森下総司, 木原慶彦, 奥田真帆, 楊印杰, 今井美沙, 枝廣陽子, 伊藤雅文, 大佐賀智, 落合友則, 三澤恭平, 大坂顯通, 小松則夫.
  • Organizer: 第30回日本サイトメトリー学会学術集会
• [Presentation] The molecular mechanism underlying the development of myeloproliferative neoplasms by mutant calreticulin (2019)
  • Author(s): Marito Araki
  • Organizer: 第81回日本血液学会学術集会
  • Invited
• [Presentation] Oncogenic activation of thrombopoietin receptor by mutant calreticulin on cell surface (2019)
  • Author(s): Marito Araki、Norio Komatsu
  • Organizer: 第78回日本癌学会学術総会
• [Presentation] calreticulin変異による骨髄増殖性腫瘍発症の分子メカニズム (2019)
  • Author(s): 荒木真理人、楊印杰、増渕菜弥、林英里奈、水上喜久、木原慶彦、今井美沙、弘中由美、橳島麻衣、枝廣陽子、大坂顯通、小松則夫
  • Organizer: 第29回日本サイトメトリー学会学術集会
  • Invited
• [Presentation] Overexpression of MPL Causes Ruxolitinib-Resistance in Myeloproliferative Neoplasms with Calreticulin Frame-Shift Mutations (2019)
  • Author(s): Shunichiro Yasuda、Satoru Aoyama、Ryoto Yoshimoto、Watanabe Daisuke、Hiroki Akiyama、Yoshihiro Umezawa、Ayako Nogami、Kouhei Yamamoto、Takeo Fujiwara、Yoko Edahiro Marito Araki、Norio Komatsu、Tetsuya Fukuda、Ayako Arai、Osamu Miura、Norihiko Kawamata
  • Organizer: 61th Annual Meeting & Exposition of American Society of Hematology
  • Int'l Joint Research
• [Presentation] Mutations in epigenetic and splicing related genes among Japanese overt and prefibrotic primary myelofibrosis using target seuencing (2019)
  • Author(s): Soji Morishita、Tomonori Ochiai、Kyohei Misawa、Tadaaki Inano、Yasutaka Fukuda、Yasumitsu Kurokawa、Yoko Edahiro、Marito Araki、Akimichi Ohsaka、Norio Komatsu
  • Organizer: The 10th JSH International Symposium
  • Int'l Joint Research
• [Presentation] Mutant calreticulin interacts with MPL in the secretion pathway for the activation on cell surface. (2019)
  • Author(s): Nami Masubuchi、Marito Araki、Yinjie Yang、Erina Hayashi、Misa Imai、Yoko Edahiro、Yumi Hironaka、Yoshihisa Mizukami、Yoshihiko Kihara、Masato Koike、Akimichi Ohsaka、Norio Komatsu
  • Organizer: 第81回日本血液学会学術集会
• [Presentation] Mutant specific sequence promotes accumulation of mutant CALR in the Golgi apparatus (2019)
  • Author(s): Yoshihisa Mizukami、Marito Araki、Misa Imai、Erina Hayashi、Nami Masubuchi、Yinjie Yang、Yoshihiko Kihara、Yumi Hironaka、Akimichi Ohsaka、Norio Komatsu
  • Organizer: 第81回日本血液学会学術集会
• [Presentation] Distinctive clinical and molecular features in absolute triple-negative essential thrombocythemia (2019)
  • Author(s): Tadaaki Inano、Marito Araki、Soji Morishita、Tomonori Ochiai、Kyohei Misawa、Yasutaka Fukuda、Naoko Midorikawa、Mai Nudejima、Masafumi Ito、Satoshi Osaga、Yoko Edahiro、Misa Imai、Teppei Taguchi、Maho Okuda、Akimichi Ohsaka、Norio Komatsu
  • Organizer: 第81回日本血液学会学術集会
• [Presentation] Establishment of a mouse model for mutant CALR myeloproliferative neoplasms (2019)
  • Author(s): Misa Imai、Yasutaka Fukuda、Teppei Taguchi、Kyohei Misawa、Honglian Tong、Kimi Araki、Motomi Osato、Akimichi Ohsaka、Marito Araki、Norio Komatsu
  • Organizer: 第81回日本血液学会学術集会
• [Presentation] 分泌経路における変異型分子シャペロンとサイトカイン受容体の会合による細胞の腫瘍化 (2019)
  • Author(s): 増渕菜弥、荒木真理人、木原慶彦、楊印杰、今井美沙、水上喜久、林英里奈、弘中由美、竹井拓、枝廣陽子、大坂顯通、小松則夫
  • Organizer: 第29回日本サイトメトリー学会学術集会
• [Presentation] 変異型特異的配列による変異型CALRの細胞内局在の規定 (2019)
  • Author(s): 水上喜久、荒木真理人、今井美沙、林英里奈、増渕菜弥、楊印杰、木原慶彦、弘中由美、大坂顯通、小松則夫
  • Organizer: 第29回日本サイトメトリー学会学術集会
• [Patent(Industrial Property Rights)] 抗切断型変異ＣＡＬＲ－ＣＤ３二重特異性抗体及び医薬組成物 (2020)
  • Inventor(s): 小松則夫、荒木真理人、木原慶彦、石田陽治、ほか
  • Industrial Property Rights Holder: 小松則夫、荒木真理人、木原慶彦、石田陽治、ほか
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2020-143551
  • Filing Date: 2020
• [Patent(Industrial Property Rights)] 切断型の変異型Ｃａｌｒｅｔｉｃｕｌｉｎに結合する抗体、及び骨髄増殖性腫瘍の診断、予防又は治療薬 (2020)
  • Inventor(s): 荒木真理人、小松則夫、木原慶彦
  • Industrial Property Rights Holder: 荒木真理人、小松則夫、木原慶彦
  • Industrial Property Rights Type: 特許
  • Filing Date: 2020
  • Overseas
• [Patent(Industrial Property Rights)] 多血症治療薬 (2020)
  • Inventor(s): 張黎臨、野村富美子、荒木真理人、小松則夫
  • Industrial Property Rights Holder: 張黎臨、野村富美子、荒木真理人、小松則夫
  • Industrial Property Rights Type: 特許
  • Filing Date: 2020
  • Overseas
• [Patent(Industrial Property Rights)] 非公開 (2020)
  • Inventor(s): 荒木真理人、小松則夫、木原慶彦
  • Industrial Property Rights Holder: 荒木真理人、小松則夫、木原慶彦
  • Industrial Property Rights Type: 特許
  • Filing Date: 2020
  • Overseas