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Myeloma

CRM1 inhibition induces tumor cell cytotoxicity and impairs osteoclastogenesis in multiple myeloma: molecular mechanisms and therapeutic implications

Abstract

The key nuclear export protein CRM1/XPO1 may represent a promising novel therapeutic target in human multiple myeloma (MM). Here we showed that chromosome region maintenance 1 (CRM1) is highly expressed in patients with MM, plasma cell leukemia cells and increased in patient cells resistant to bortezomib treatment. CRM1 expression also correlates with increased lytic bone and shorter survival. Importantly, CRM1 knockdown inhibits MM cell viability. Novel, oral, irreversible selective inhibitors of nuclear export (SINEs) targeting CRM1 (KPT-185, KPT-330) induce cytotoxicity against MM cells (ED50<200 nM), alone and cocultured with bone marrow stromal cells (BMSCs) or osteoclasts (OC). SINEs trigger nuclear accumulation of multiple CRM1 cargo tumor suppressor proteins followed by growth arrest and apoptosis in MM cells. They further block c-myc, Mcl-1, and nuclear factor κB (NF-κB) activity. SINEs induce proteasome-dependent CRM1 protein degradation; concurrently, they upregulate CRM1, p53-targeted, apoptosis-related, anti-inflammatory and stress-related gene transcripts in MM cells. In SCID mice with diffuse human MM bone lesions, SINEs show strong anti-MM activity, inhibit MM-induced bone lysis and prolong survival. Moreover, SINEs directly impair osteoclastogenesis and bone resorption via blockade of RANKL-induced NF-κB and NFATc1, with minimal impact on osteoblasts and BMSCs. These results support clinical development of SINE CRM1 antagonists to improve patient outcome in MM.

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Acknowledgements

We are grateful for Hao Wang, Matt Ma, Michelle Chen, Yiguo Hu and Sunyoung Kong for excellent technical support to the project. We thank the input from Haoqiang Ying and Dilara McCauley from Karyopharm Pharmaceutics. We also thank the nursing staff and clinical research coordinators of the LeBow Institute for Myeloma Therapeutics and the Jerome Lipper Multiple Myeloma Center of the Dana-Farber Cancer Institute for support and help in providing primary tumor specimens for this study. This study was supported by National Institutes of Health Grants RO-1 50947, PO1–78378 and DF/HCC SPORE in Multiple Myeloma P50CA100707; and the Lebow Fund to Cure Myeloma (KCA); KCA is an American Cancer Society Clinical Research Professor.

Author Contributions

Y-TT, YL, SS, MK, KCA conceptualized research and formed the hypothesis of this paper; Y-TT, YL, YC, SS, ALK designed, performed experiments, analyzed data; FZ, AC, AAM analyzed expression data; CA, MC, YC, MYZ, AC, MR, DT, J-RS-M, TK, YG, WS, IG, LW, performed the in vitro research and collected data; SS, ALK, CA, MC, MYZ designed, performed and analyzed animal work; YL, YC, SS, MK, LW, SAS provided reagents, analytic tools and input to studies; NCM, PR, KCA provided MM patient samples; Y-TT wrote the manuscript; Y-TT, SS, MK, KCA critically evaluated and edited the manuscript.

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Correspondence to Y-T Tai.

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YL, WS, J-RS-M, TK, SS, MK are employees of Karyopharm Therapeutics Inc., whose product was used in this research PR serves on advisory boards to Millennium, Celgene, Novartis, Johnson & Johnson and Bristol-Myers Squibb NCM serves on advisory boards to Millennium, Celgene and Novartis KCA serves on advisory boards to Onyx, Celgene, Gilead and Sanofi-Aventis. The remaining authors declare no conflict of interest.

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Tai, YT., Landesman, Y., Acharya, C. et al. CRM1 inhibition induces tumor cell cytotoxicity and impairs osteoclastogenesis in multiple myeloma: molecular mechanisms and therapeutic implications. Leukemia 28, 155–165 (2014). https://doi.org/10.1038/leu.2013.115

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Keywords

  • multiple myeloma (MM)
  • selective inhibitors of nuclear export (SINEs) against CRM1/XPO1
  • osteoclasts (OC)
  • nuclear factor-κB (NF-κB) activation
  • nuclear export protein
  • tumor suppressors

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