基礎研究

リンパ系腫瘍グループ

血液がんの中でも特に診断及び治療が難しい病気の一つが、悪性リンパ腫です。特にT細胞性リンパ腫は既存の治療法に抵抗性で、あるいは、一度は治療が奏功しても高頻度に再発をきたします。この現状を変えて行くために、新しい治療標的分子の同定、分子標的薬の開発が望まれていますが、その基盤となるべき分子病態の理解は遅れています。

近年は次世代シークエンサーの登場により網羅的な体細胞遺伝子変異解析が可能になりました。T細胞性リンパ腫の分野でも腫瘍形成に重要と考えられる遺伝子変異が多数報告されてきています(Nakagawa et al. JEM 211:2497-505, 2014.など)。

それらの遺伝子変異の発見が新しい治療法開発につながると良いのですが、実際はそう簡単ではないこともわかってきています。その遺伝子変異はリンパ腫ができ始めたきっかけだったかもしれませんが、病気が発見され治療が必要な時にはすでに不必要になっているかもしれません。また、がんは一つの遺伝子異常ではなく、たくさんの遺伝子群によるnet effectで腫瘍形質を維持しています。一つの遺伝子を標的としても、不十分な効果しか得られないこともあります。では、どの遺伝子を標的にすれば本当に治療効果をあげられるのでしょうか？これは全ての悪性リンパ腫患者さん、そして、その治療に関わる人々にとって、最も重要な問題ではないでしょうか？

この重要問題に正面から取り組むために、我々はCRISPR/Cas9網羅的遺伝子ノックアウトスクリーニングを基盤とした仕事を進めています。CRISPR/Cas9は近年開発されたばかりの革新的ゲノム編集技術であり、これを用いることで従来法より飛躍的に高い効率・特異性で標的遺伝子をノックアウトすることが可能になってきました。我々はこのシステムをヒトT細胞性リンパ腫細胞株に導入、約 20000遺伝子を網羅的に解析し、どの遺伝子を標的とすることでT細胞リンパ腫細胞を根絶できるのかを探っています（図）。

代表的論文

1. Nakagawa, M., Shaffer, AL., Ceribelli, M., Zhang, M., Wright, GW., Huang, da W., Xiao, W., Powell, J., Petrus, MN., Yang, Y., Phelan, JD., Kohlhammer, H., Dubois, SP., Yoo, HM., Bachy, E., Webster, DE., Yang, Y., Xu, W., Yu, X., Zhao, H., Bryant, BR., Shimono, J., Ishio, T., Maeda, M., Green, PL., Waldmann, TA. and Staudt, LM. Targeting the HTLV-I-regulated BATF3/IRF4 Transcriptional Network in Adult T-Cell Leukemia/Lymphoma. Cancer Cell, 34:286-297, 2018.
   
   shRNAライブラリースクリーニングとCRISPRライブラリースクリーニングを駆使してT細胞性リンパ腫の一病型である成人T細胞白血病リンパ腫においてBATF3/IRF4が重要な治療標的であることを見出しました。成人T細胞白血病リンパ腫の原因ウイルスであるHTLV-IがBATF3の発現に関わることで、ウイルスと腫瘍発生の関係性についても明らかにしました。
   
   
2. Ishio, T., Kumar, S., Shimono, J., Daenthanasanmak, A., Dubois, S., Lin, Y., Bryant, BR., Petrus, MN., Bachy, E., Huang, DW., Yang, Y., Green, PL., Hasegawa, H., Maeda, M., Goto, H., Endo, T., Yokota, T., Hatanaka, KC., Hatanaka, Y., Tanaka, S., Matsuno, Y., Yang, Y., Hashino, S., Teshima, T., Waldmann, TA., Staudt, LM., Nakagawa, M. Genome-wide CRISPR screen identifies CDK6 as a therapeutic target in Adult T-cell leukemia/lymphoma. Blood. 139:1541-1556, 2022.
   
   石尾先生が大学院生としてCRISPRライブラリースクリーニングの解析を進めてくださり、成人T細胞白血病リンパ腫においてCDK6が重要な治療標的であることを見出しました。CDK6はキナーゼであり、その阻害薬は成人T細胞白血病リンパ腫に有効である可能性を指摘しました。
   
   
3. Chiba, M., Shimono, J., Ishio, T., Takei, N., Kasahara, K., Ogasawara, R., Ara, T., Goto, H., Izumiyama, K., Otsuguro, S., Perera, LP., Hasegawa, H., Maeda, M., Hashino, S., Maenaka, K., Teshima, T., Waldmann, TA., Yang, Y., Nakagawa, M. Genome-wide CRISPR screens identify CD48 defining susceptibility to NK cytotoxicity in peripheral T-cell lymphomas. Blood. 140:1951-1963, 2022.
   
   千葉先生が大学院生として解析して下さった仕事です。成人T細胞白血病リンパ腫はCD48という分子の発現を減少させることで、NK細胞による腫瘍監視機構から逃れることを見出しました。

獲得競争的研究費

• 2017年　公益財団法人 先進医薬研究振興財団　血液医学分野研究助成金
• 2017年　公益財団法人 武田科学振興財団　医学系研究奨励
• 2018年　日本学術振興会科学研究費助成事業　基盤研究(C)
• 2018年　日本白血病研究基金 一般研究賞
• 2018年　寿原記念財団 研究助成
• 2018年　高松宮妃癌研究基金　研究助成金
• 2019年　公益財団法人 アステラス病態代謝研究会　研究助成金
• 2021年　日本学術振興会科学研究費助成事業　基盤研究(B)

Lymphoid Malignancies Group

RESEARCH INTERESTS

Functional genomics for discovering the therapeutic molecular targets in PTCL

> Identifying the "Achilles' heel" genes in peripheral T-cell lymphomas as molecular targets for therapeutic intervention.
> Defining the molecular mechanisms of frequently mutated CCR4 gene in PTCL.

Overview

Human peripheral T-cell lymphomas (PTCL) is approximately 10 % of non-Hodgkin Lymphomas in worldwide. PTCL comprises heterogeneous diseases, where most are aggressive with less than 40% of 5-year overall survival with current treatment strategy. A discovery for new therapeutic modalities is urgently need.

Recently, the Next generation sequencing technology enabled us to find frequently mutated genes in PTCL. In my previous work as a postdoctoral fellow in Lou Staudt Lab and Thomas Waldmann lab, we discovered frequent gain-of-function mutation in chemokine receptor CCR4 gene in Adult T-cell leukemia/lymphoma (ATLL) and provided functional evidences that inhibition of CCR4 signaling might have therapeutic potential for patients with ATLL. However most of other somatically mutated genes in PTCL were not fully characterized in terms of therapeutic targets.
On the other hand, it has been recently known that the malignant cells potentially acquire the dependency on un-mutated genes which is the phenomenon called as "non-oncogene addiction". These observations indicate that non-biased and comprehensive functional investigation should be conducted in order to discover the therapeutic molecular targets in PTCL.

The goal of our laboratory is to understand the oncogenic molecular networks in PTCL which can be exploited for therapeutic intervention. 

PUBLICATION LIST

1. Wei, W., Song, Z., Chiba, M., Wu, W., Jeong, S., Zhang, JP., Kadin, ME., Nakagawa, M., Yang, Y. Analysis and therapeutic targeting of the EP300 and CREBBP acetyltransferases in anaplastic large cell lymphoma and Hodgkin lymphoma. Leukemia. 37:396-407, 2022.   
2. Chiba, M., Shimono, J., Ishio, T., Takei, N., Kasahara, K., Ogasawara, R., Ara, T., Goto, H., Izumiyama, K., Otsuguro, S., Perera, LP., Hasegawa, H., Maeda, M., Hashino, S., Maenaka, K., Teshima, T., Waldmann, TA., Yang, Y., Nakagawa, M. Genome-wide CRISPR screens identify CD48 defining susceptibility to NK cytotoxicity in peripheral T-cell lymphomas. Blood. 140:1951-1963, 2022.
3. Ishio, T., Kumar, S., Shimono, J., Daenthanasanmak, A., Dubois, S., Lin, Y., Bryant, BR., Petrus, MN., Bachy, E., Huang, DW., Yang, Y., Green, PL., Hasegawa, H., Maeda, M., Goto, H., Endo, T., Yokota, T., Hatanaka, KC., Hatanaka, Y., Tanaka, S., Matsuno, Y., Yang, Y., Hashino, S., Teshima, T., Waldmann, TA., Staudt, LM., Nakagawa, M. Genome-wide CRISPR screen identifies CDK6 as a therapeutic target in Adult T-cell leukemia/lymphoma. Blood. 139:1541-1556, 2022.
4. Wang, H., Wei, W., Zhang, JP., Song, Z., Li, Y., Xiao, W., Liu, Y., Zeng, MS., Petrus, MN., Thomas, CJ., Kadin, ME., Nakagawa, M., Waldmann, TA., Yang, Y. A novel model of alternative NF-kappaB pathway activation in anaplastic large cell lymphoma. Leukemia, 35:1976-1989, 2021.
5. Zhang, JP., Song, Z., Wang, HB., Lang, L., Yang YZ., Xiao, W., Webster, DE., Wei, W., Barta, SK., Kadin, ME., Staudt, LM., Nakagawa, M., and Yang, Y. A novel model of controlling PD-L1 expression in ALK+ Anaplastic Large Cell Lymphoma. Blood, 132:171-185, 2019. (Co-corresponding author)  
6. Nakagawa, M., Shaffer, AL., Ceribelli, M., Zhang, M., Wright, GW., Huang, da W., Xiao, W., Powell, J., Petrus, MN., Yang, Y., Phelan, JD., Kohlhammer, H., Dubois, SP., Yoo, HM., Bachy, E., Webster, DE., Yang, Y., Xu, W., Yu, X., Zhao, H., Bryant, BR., Shimono, J., Ishio, T., Maeda, M., Green, PL., Waldmann, TA. and Staudt, LM. Targeting the HTLV-I-regulated BATF3/IRF4 Transcriptional Network in Adult T-Cell Leukemia/Lymphoma. Cancer Cell, 34:286-297, 2018.
7. Phelan, JD., Young, RM., Webster, DE., Roulland, S., Wright, GW., Kasbekar, M., Shaffer, AL., Ceribelli, M., Wang, JQ., Schmitz, R., Nakagawa, M., Bachy, E., Huang, da W., Ji, Y., Chen, L., Yang, Y., Zhao, H., Yu, X., Xu, W., Palisoc, MM., Valadez, RR., Davies-Hill, T., Wilson, WH., Chan, WC., Jaffe, ES., Gascoyne, RD, Campo, E., Rosenwald, A., Ott, G., Delabie, J., Rimsza, LM, Rodriguez, FJ., Estephan, F., Holdhoff, M., Kruhlak, MJ., Hewitt, SM., Thomas, CJ., Pittaluga, S, Oellerich, T., and Staudt, LM. A Multiprotein Supercomplex Controlling Oncogenic Signaling in Lymphoma. Nature, 2018. In Press.
8. Perera, LP., Zhang, M., Nakagawa, M., Petrus, MN., Maeda, M., Kadin, ME., Waldmann, TA., Perera, PY. Chimeric antigen receptor modified T cells that target chemokine receptor CCR4 as a therapeutic modality for T-cell malignancies. American Journal of Hematology, 2017. In Press.
9. Shimono, J., Miyoshi, H., Kiyasu, J., Sato, K., Kamimura, T., Eto, T., Miyagishima, T., Nagafuji, K., Teshima, T., Ohshima, K. Clinicopathologic analysis of primary splenic diffuse large B-cell lymphoma. Br J Haemtol 2017 In Press.
10. Shimono, J., Miyoshi, H., Seto, M., Ohshima, K. Clinical features of diffuse large B-cell lymphoma with polyploidy. Pathol Int, 67:17-23, 2017. 
11. Chen, J., Zhang. Y,, Petrus, MN., Xiao, W., Nicolae, A., Raffeld, M., Pittaluga, S., Bamford, RN., Nakagawa, M., Ouyanga, S., Epsteine, AL., Kadin, ME., Del Mistro, A., Woessnerh, R., Jaffe, ES., Waldmann, TA. Cytokine receptor signaling is required for the survival of ALK- anaplastic large cell lymphoma even in the presence of JAK1/STAT3 mutations. Proc. Natl. Acad. Sci. U S A, 2017. In Press. 
12. Yang, Y., Kelly, P., Shaffer, AL., Schmitz, R., Yoo, HM., Liu, X., Huang, da W., Webster, D., Young, RM., Nakagawa, M., Ceribelli, M., Wright, GW., Yang, Y., Zhao, H., Yu, X., Xu, W., Chan, WC., Jaffe, ES., Gascoyne, RD., Campo, E., Rosenwald, A, Ott, G., Delabie, J., Rimsza, L., Staudt, LM.　 Targeting Non-proteolytic Protein Ubiquitination for the Treatment of Diffuse Large B Cell Lymphoma.　 Cancer Cell, 29:494-507, 2016.
13. Hodson, DJ., Shaffer, AL., Xiao, W., Wright, GW., Schmitz, R., Phelan, JD., Yang, Y., Webster, DE., Rui, L., Kohlhammer, H., Nakagawa, M., Waldmann, TA., Staudt, LM. Regulation of normal B-cell differentiation and malignant B-cell survival by OCT2. Proc. Natl. Acad. Sci. U S A, 113:E2039-46, 2016
14. Miyoshi, H., Kiyasu, J., Kato, T., Yoshida, N., Shimono, J., Yokoyama, S., Taniguchi, H., Sasaki, Y., Kurita, D., Kawamoto, K., Kato, K., Imaizumi, Y., Seto, M., Ohshima, K. PD-L1 expression on neoplastic or stromal cells is respectively a poor or good prognostic factor for adult T-cell leukemia/lymphoma. Blood, 128:1374-81, 2016. 
15. Yokoyama, S., Miyoshi, H., Nakashima, K., Shimono, J., Hashiguchi, T., Mitsuoka, M., Takamori, S., Akagi, Y., Ohshima, K. Prognostic Value of Programmed Death Ligand 1 and Programmed Death 1 Expression in Thymic Carcinoma. Clin Cancer Res, 22:4727-34, 2016
16. Yoshida, N., Miyoshi, H., Kato, T., Sakata-Yanagimoto, M., Niino, D., Taniguchi, H., Moriuchi, Y., Miyahara, M., Kurita, D., Sasaki, Y., Shimono, J., Kawamoto, K., Utsunomiya, A., Imaizumi, Y., Seto, M., Ohshima, K. CCR4 frameshift mutation identifies a distinct group of adult T cell leukaemia/lymphoma with poor prognosis. J Pathol, 238:621-6, 2016.
17. Nakagawa, M., Schmitz, R., Xiao, W., Goldman, CK., Yang, Y., Xu, W., Yu, X., Waldmann, TA., Staudt, LM. Gain-of-Function CCR4 Mutations in Adult T-cell Leukemia/Lymphoma. J. Exp. Med., 211:2497-505, 2014.
18. Yu, P., Petrus, MN., Ju, W., Zhang, M., Conlon, KC., Nakagawa, M., Maeda, M., Bamford, RN., Waldmann, TA. Augmented efficacy with the combination of blockade of the Notch-1 pathway, bortezomib and romidepsin in a murine MT-1 adult T-cell leukemia model. Leukemia, 29:556-66, 2014.
19. Suguro, M., Yoshida, N., Umino, A., Kato, H., Tagawa, H., Nakagawa, M., Fukuhara, N., Karnan, S., Takeuchi, I., Hocking, TD., Arita, K., Karube, K., Tsuzuki, S., Nakamura, S., Kinoshita, T., Seto, M. Clonal heterogeneity of lymphoid malignancies correlates with poor prognosis. Cancer Sci., 105:897-904, 2014.
20. Karube, K., Nakagawa, M., Tsuzuki, S., Takeuchi, I., Honma, K., Nakashima, Y., Shimizu, N., Ko, YH., Morishima, Y., Ohshima, K., Nakamura, S., Seto, M. Identification of FOXO3 and PRDM1 as tumor-suppressor gene candidates in NK-cell neoplasms by genomic and functional analyses. Blood, 118: 3195-204, 2011.
21. Nakagawa, M., Tsuzuki, S, Honma, K., Taguchi, O., Seto, M. Synergistic effect of Bcl2, Myc and Ccnd1 transforms mouse primary B-cells into malignant cells. Haematologica, 96: 1318-1326, 2011. 
22. Umino, A., Nakagawa, M., Utsunomiya, A., Tsukasaki, K., Taira, N., Katayama, N., Seto, M. Clonal evolution of adult T-cell leukemia/lymphoma takes place in lymph node. Blood, 117: 5473-5478, 2011. 
23. Kato, H., Kagami, Y., Nakagawa, M., Karnan, S., Yatabe, Y., Nakamura, S., Morishima, Y., Seto, M. IL-4/CD40L Co-Stimulation Induces Long-Term Proliferation for CD10-Positive Germinal Center B Cell-Like Diffuse Large B-Cell Lymphoma. The Open Leukemia Journal, 3: 60-68, 2010.
24. Seto, M., Honma, K., Nakagawa, M. Diversity of genome profiles in malignant lymphoma. Cancer Sci., 101: 573-578, 2010. 
25. Honma, K. Tsuzuki, S. Nakagawa,　M., Tagawa, H., Nakamura, S., Morishima, Y., Seto, M. TNFAIP3/A20 functions as a novel tumor suppressor gene in several subtypes of non-Hodgkin lymphomas. Blood, 114: 2467-2475, 2009. 
26. Nakagawa, M., Nakagawa-Oshiro, A., Karnan, S., Tagawa, H., Utsunomiya, A., Nakamura, S., Takeuchi, I., Ohshima, K., Seto, M. Array CGH analysis of PTCL-U reveals a distinct subgroup with genetic alterations similar to lymphoma-type ATLL. Clin. Cancer Res., 15: 30-38, 2009.
27. Takeuchi, I., Tagawa, H., Tsujikawa, A., Nakagawa, M., Katayama-Suguro, M., Guo, Y., Seto, M. The potential of copy number gains and losses, detected by array-based comparative genomic hybridization, for computational differential diagnosis of B-cell lymphomas and genetic regions involved in lymphomagenesis. Haematologica, 94: 61-69, 2009.
28. Hashino, S., Kobayashi, S., Takahata, M., Onozawa, M., Nakagawa, M., Kawamura, T., Fujisawa, F., Izumiyama, K., Kahata, K., Kondo, T., Asaka, M. Graft-versus-tumor effect after reduced-intensity allogeneic hematopoietic stem cell transplantation in a patient with advanced colon cancer. Int. J. Clin. Oncol., 13: 176-180, 2008. 
29. Honma, K., Tsuzuki, S., Nakagawa, M., Karnan, S., Aizawa, Y., Kim, WS., Kim, YD., Ko, YH., Seto, M. TNFAIP3 is the target gene of chromosome band 6q23.3-q24.1 loss in ocular adnexal marginal zone B cell lymphoma. Genes Chrom. Cancer, 47: 1-7, 2008. 
30. Hashino, S., Morita, L., Takahata, M., Onozawa, M., Nakagawa, M., Kawamura, T., Fujisawa, F., Kahata, K., Izumiyama, K., Yonezumi, M., Chiba, K., Kondo, T., Asaka, M. Administration of micafungin as prophylactic antifungal therapy in patients undergoing allogeneic stem cell transplantation. Int. J. Hematol., 87: 91-97, 2008. 
31. Nakagawa, M., Hashino, S., Takahata, M., Kawamura, T., Fujisawa, F., Kahata, K., Kondo, T., Imamura, M., Ando, S., Asaka, M. Successful reduced-intensity stem cell transplantation with cord blood for a poor-prognosis adult with refractory chronic active epstein-barr virus infection. Int. J. Hematol., 85: 443-445, 2007.
32. Fukuhara, N., Nakamura, T., Nakagawa, M., Tagawa, H., Takeuchi, I., Yatabe, Y., Morishima, Y., Nakamura, S., Seto, M. Chromosomal imbalances are associated with outcome of Helicobacter pylori eradication in t(11;18)(q21;q21) negative gastric MALT lymphomas. Genes Chrom. Cancer, 46: 784-790, 2007.
33. Onozawa, M., Hashino, S., Takahata, M., Fujisaw, F., Kawamura, T., Nakagawa, M., Kahata, K., Kondo, T., Ota, S., Tanaka, J., Imamura, M., Asaka, M. Relationship between preexisting anti-varicella-zoster virus (VZV) antibody and clinical VZV reactivation in hematopoietic stem cell transplantation recipients. J. Clin. Microbiol., 44: 4441-4443, 2006. 
34. Nakagawa, M., Seto, M., Hosokawa, Y. Molecular pathogenesis of MALT lymphoma: two signaling pathways underlying anti-apoptotic effect of API2-MALT1 fusion protein. Leukemia, 20: 929-936, 2006. 
35. Nakagawa, M., Seto, M., Hosokawa, Y. Molecular pathogenesis of MALT lymphoma: two signaling pathways underlying anti-apoptotic effect of API2-MALT1 fusion protein. Leukemia, 20: 929-936, 2006.
36. Hosokawa, Y., Suzuki, H., Nakagawa, M., Lee, T.H., Seto, M. API2-MALT1 fusion protein induces transcriptional activation of the API2 gene through NF-kappaB binding elements: Evidence for a positive feed-back loop pathway resulting in unremitting NF-kappaB activation. Biochem. Biophys. Res. Commun., 334: 51-60, 2005.
37. Nakagawa, M., Kameoka, Y., Suzuki, R. Nucleophosmin in acute myelogenous leukemia. N. Engl. J. Med., 352: 1819-1820, 2005. 
38. Izumiyama, K., Nakagawa, M., Yonezumi, M., Kasugai, Y., Suzuki, R., Suzuki, H., Tsuzuki, S., Hosokawa, Y., Asaka, M., Seto, M. Stability and subcellular localization of API2-MALT1 chimeric protein involved in t(11;18) (q21;q21) MALT lymphoma. Oncogene, 22: 8085-8092, 2003.
39. Nakagawa, M., Miyagishima, T., Kamata, T., Arai, S., Miura, Y., Onishi, S., Kishimoto, A., Kamishima, Y., Choi, G.H., Kudo, M., Okabe, M. Refractory idiopathic cold agglutinin disease successfully treated with intermittent high-dose cyclophosphamide. Rinsho Ketsueki, 42: 713-715, 2001.