Data from Targeting Autophagy Augments <i>In Vitro</i> and <i>In Vivo</i> Antimyeloma Activity of DNA-Damaging Chemotherapy

Abstract

<div>Abstract<p><b>Purpose:</b> Although autophagy occurs in most tumor cells following DNA damage, it is still a mystery how this DNA-damaging event turns on the autophagy machinery in multiple myeloma (MM) and how the functional status of autophagy impacts on its susceptibility to death in response to DNA-damaging chemotherapy.</p><p><b>Experimental Design:</b> We investigate the effects of DNA damage on autophagy in MM cells and elucidate its underlying molecular mechanism. Then, we examined the impacts of pharmacologic or genetic inhibition of autophagy on DNA damage–induced apoptosis. Furthermore, the antimyeloma activity of autophagy inhibitor in combination with DNA-damaging agents was evaluated in MM xenograft models.</p><p><b>Results:</b> We showed that DNA-damaging drugs, doxorubicin and melphalan, induce caspase-dependent apoptosis and concurrently trigger Beclin 1–regulated autophagy in human MM cell lines H929 and RPMI 8226. Mechanistically, association of autophagy execution proteins Beclin 1 with class III phosphoinositide 3-kinase, which is inhibited by Bcl-2 recruitment, contributes directly to the autophagic process. Importantly, targeting suppression of autophagy by minimally toxic concentrations of pharmacologic inhibitors (hydroxychloroquine and 3-methyladenine) or short hairpin RNAs against autophagy genes, <i>Beclin 1</i> and <i>Atg5</i>, dramatically augments proapoptotic activity of DNA-damaging chemotherapy both <i>in vitro</i> using MM cell lines or purified patient MM cells and <i>in vivo</i> in a human plasmacytoma xenograft mouse model.</p><p><b>Conclusion:</b> These data can help unravel the underlying molecular mechanism of autophagy in DNA-damaged MM cells and also provide a rationale for clinical evaluation of autophagy inhibitors in combination with DNA-damaging chemotherapy in MM. <i>Clin Cancer Res; 17(10); 3248–58. ©2011 AACR</i>.</p></div>