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Importin β1 regulates cell growth and survival during adult T cell leukemia/lymphoma therapy

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Summary

There is no cure for adult T cell leukemia/lymphoma (ATLL) associated with human T cell leukemia virus type 1 (HTLV-1), and novel targeted strategies are needed. NF-κB and AP-1 are crucial for ATLL, and both are transported to the nucleus by an importin (IPO)α/β heterodimeric complex to activate target genes. In this study, we aimed to elucidate the function of IPOβ1 in ATLL. The expression of IPOβ1 was analyzed by western blotting and RT-PCR. Cell growth, viability, cell cycle, apoptosis and intracellular signaling cascades were examined by the water-soluble tetrazolium-8 assay, flow cytometry and western blotting. Xenograft tumors in severe combined immune deficient mice were used to evaluate the growth of ATLL cells in vivo. IPOβ1 was upregulated in HTLV-1-infected T cell lines. Further, IPOβ1 knockdown or the IPOβ1 inhibitor importazole and the IPOα/β1 inhibitor ivermectin reduced HTLV-1-infected T cell proliferation. However, the effect of inhibitors on uninfected T cells was less pronounced. Further, in HTLV-1-infected T cell lines, inhibitors suppressed NF-κB and AP-1 nuclear transport and DNA binding, induced apoptosis and poly (ADP-ribose) polymerase cleavage, and activated caspase-3, caspase-8 and caspase-9. Inhibitors also mediated G1 cell cycle arrest. Moreover, the expression of NF-κB- and AP-1-target proteins involved in cell cycle and apoptosis was reduced. In vivo, the IPOα/β1 inhibitor ivermectin decreased ATLL tumor burden without side effects. IPOβ1 mediated NF-κB and AP-1 translocation into ATLL cell nuclei, thereby regulating cell growth and survival, which provides new insights for targeted ATLL therapies. Thus, ivermectin, an anti-strongyloidiasis medication, could be a potent anti-ATLL agent.

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Acknowledgments

The authors would like to thank Fujisaki Cell Center, Hayashibara Biochemical Laboratories, Inc. for providing C5/MJ, HUT-102 and MT-1 cells, Dr. Naoki Yamamoto (Tokyo Medical and Dental University) for providing MT-2 and MT-4 cells, Dr. Diane Prager (UCLA School of Medicine) for providing SLB-1 cells, Dr. Michiyuki Maeda (Kyoto University) for providing ED-40515(−) cells, and Dr. Masahiro Fujii (Niigata University) for providing TL-OmI cells. Recombinant human IL-2 was kindly provided by Takeda Pharmaceutical Company Ltd. The measurement of protein concentrations was performed at the University of the Ryukyus Center for Research Advancement and Collaboration. We would like to thank Editage (www.editage.jp) for English language editing.

Funding

This study was supported partially by JSPS KAKENHI (17 K07175).

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Authors and Affiliations

  1. Department of Microbiology and Oncology, Graduate School of Medicine, University of the Ryukyus, 207 Uehara, Nishihara, Okinawa, 903-0215, Japan

    Chie Ishikawa & Naoki Mori

  2. Division of Health Sciences, Transdisciplinary Research Organization for Subtropics and Island Studies, University of the Ryukyus, Nishihara, Okinawa, Japan

    Chie Ishikawa

  3. Department of Pathology, Institute of Tropical Medicine, Nagasaki University, Nagasaki, Japan

    Masachika Senba

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  1. Chie Ishikawa
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Correspondence to Naoki Mori.

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All animal experiments were approved by the Animal Care and Use Committee of University of the Ryukyus (A2016149).

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Ishikawa, C., Senba, M. & Mori, N. Importin β1 regulates cell growth and survival during adult T cell leukemia/lymphoma therapy. Invest New Drugs 39, 317–329 (2021). https://doi.org/10.1007/s10637-020-01007-z

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