Abstract 2142: The nuclear export protein CRM1 (XPO1) regulates multiple myeloma cell growth, osteoclastogenesis, and myeloma-induced osteolysis.
Authors: Tai Yu-Tzu , Landesman Yosef , Acharya Chirag , Calle Yolanda , Zhong Mike , Cea Michele , Tannenbaum Daniel , Cagnetta Antonia , Reagan Michaela , Munshi Aditya , Senapedis William , Saint-Martin Jean-Richard , Kashyap Trinayan , Shacham Sharon , Kauffman Michael , Gu Yumei , Wu Lizi , Schey Steve , Ghobrial Irene , Kung Andrew , Munshi Nikhil , Richardson Paul , Anderson Kenneth , Ying Haoqiang 
Published: August 2014

Abstract
The key nuclear export protein CRM1 (chromosomal region maintenance 1, Exportin 1, XPO1) may directly contribute to the pathophysiology of human multiple myeloma (MM). Here, we characterized the role of CRM1 in MM biology and defined the efficacy and molecular mechanisms of novel oral, irreversible, selective inhibitors of nuclear export (SINEs) targeting CRM1 against MM. CRM1 is significantly elevated in patient MM vs. normal plasma cells at transcript and protein levels. CRM1 downregulation by shCRM1 lentiviruses inhibited cell growth and survival of MM cells (p&lt; 0.01). Importantly, SINEs (KPT-185, KPT-251, KPT-276, and KPT-330) blocked proliferation and decreased survival of MM cell lines and patient MM cells (LD50 2-log differences). SINEs potently enhanced nuclear accumulation of multiple CRM1 cargo tumor suppressor proteins, including rapid induction of p53 and IκB (as early as 2h after drug treatment) followed by FOXO1A, FOXO3A, p27, and PP2A in MM cells. Transcripts of p53 and its downstream targets (p21, PUMA, BAX) were induced by KPT-185, thereby inducing strong growth arrest and apoptosis. KPT-185 decreased MM oncogenes (c-myc, c-maf), anti-apoptosis molecules Mcl-1 and Bcl-xL; increased pro-apoptotic protein BAX; as well as inhibited pIκBα. Inhibition of pIκBα further correlated with KPT-185-blocked NFκB p65 DNA-binding activity in MM cells with or without A Proliferation-Inducing Ligand (APRIL) stimulation. KPT-185 further downregulated CRM1 protein, which was blocked by bortezomib; concurrently, KPT-185 (or KPT-330) upregulated CRM1 mRNA in MM cells. Cleavage of caspase 3 and PARP was markedly increased in MM1R cells treated with KPT-185 and bortezomib vs. either drug alone, confirming enhanced cytotoxicity by combination of these agents. Combined dex with KPT-185 (or KPT-276) induced synergistic cytotoxicity against MM cells (combination indices &lt; 1). Moreover, KPT-185 and KPT-330 impaired osteoclastogenesis and bone resorption via blockade of RANKL-induced NFκB activation and NFATc1 in OC precursor cells, without impacting osteoblasts and BMSCs. Finally, SINEs (KPT-251 and KPT-276) suppressed MM cell growth (p&lt; 0.01), diminished MM cell-induced osteolysis, and prolonged survival of SCID mice with diffuse human MM bone lesions. Together, these results identify CRM1 as a promising novel target in MM, strongly supporting clinical development of SINE CRM1 inhibitors to inhibit both MM cell growth and related bone disease. The oral SINE KPT-330 is ongoing in MM and other hematological malignancies.
Citation Format: Yu-Tzu Tai, Yosef Landesman, Chirag Acharya, Yolanda Calle, Mike Zhong, Michele Cea, Daniel Tannenbaum, Antonia Cagnetta, Michaela Reagan, Aditya Munshi, William Senapedis, Jean-Richard Saint-Martin, Trinayan Kashyap, Sharon Shacham, Michael Kauffman, Yumei Gu, Lizi Wu, Steve Schey, Irene Ghobrial, Andrew Kung, Nikhil Munshi, Paul Richardson, Kenneth Anderson, Haoqiang Ying. The nuclear export protein CRM1 (XPO1) regulates multiple myeloma cell growth, osteoclastogenesis, and myeloma-induced osteolysis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2142. doi:10.1158/1538-7445.AM2013-2142