Niche-based screening identifies small-molecule inhibitors of leukemia stem cells

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Efforts to develop more effective therapies for acute leukemia may benefit from high-throughput screening systems that reflect the complex physiology of the disease, including leukemia stem cells (LSCs) and supportive interactions with the bone marrow microenvironment. The therapeutic targeting of LSCs is challenging because LSCs are highly similar to normal hematopoietic stem and progenitor cells (HSPCs) and are protected by stromal cells in vivo. We screened 14,718 compounds in a leukemia-stroma co-culture system for inhibition of cobblestone formation, a cellular behavior associated with stem-cell function. Among those compounds that inhibited malignant cells but spared HSPCs was the cholesterol-lowering drug lovastatin. Lovastatin showed anti-LSC activity in vitro and in an in vivo bone marrow transplantation model. Mechanistic studies demonstrated that the effect was on target, via inhibition of HMG-CoA reductase. These results illustrate the power of merging physiologically relevant models with high-throughput screening.

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Figure 1: A high-throughput system for probing LSCe cells within a stromal niche.
Figure 2: New small molecule BRD7116 selectively targets LSCe cells by cell-autonomous and cell-non-autonomous mechanisms.
Figure 3: Lovastatin selectively inhibits mouse and human leukemia cells in co-culture.
Figure 4: Sensitivity of LSCe cells to HMGCR inhibition.

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We wish to thank L. VerPlank, V. Raksakulthai, A. Liberzon, M. Palmer, G. Cowley, J. Burbank, P. Aspesi Jr., M. Bliss-Moreau, D. Barker, B. Wagner, A. Krivtsov, S.A. Armstrong, R. Karmacharya, J. Perez, R. Onofrio, D. Thomas, R. Busanelli, D. Auclair, K. Masson, J. Du, C. Moore, M. Kharas, Y. Hoshida, D. Bachovchin, A. Mullally, J. Grabarek, A. Fraser, A. Basu, J. Cheah, N. Bodycombe, C. Mulrooney, S. Johnston, G. Walzer, D. Wilpitz, A. Bracha, A. Fabian, C. Hon, J. McGrath, C. Hartland, M. Hickey, T.R. Jones, M. Bray, K. Sokolnicki, R. Okabe, M. Paktinat, M. McConkey, L. Gaffney, L. Solomon, K. Rose and Broad Compound Management. We also thank the Golub, Gilliland and Ebert laboratories for scientific discussions and technical expertise. This work was funded by grants from the Starr Cancer Consortium (SCC Award l1-A50), the US National Institutes of Health (U54CA112962, U01HL1004402, T32 HL007623, T32 GM007753, N01-CO-12400, R01 GM089652, RL1CA133834, 20XS139, RL1HG004671, RL1CA133834, RL1GM084437 and UL1RR024924) and the US National Science Foundation (DBI 1148823). The content of this publication is solely the responsibility of the authors and does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products or organizations imply endorsement by the US Government. T.R.G., D.G.G. and S.L.S. are investigators at the Howard Hughes Medical Institute.

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A.E.C., A.F.S., A.L.S., A.M.S., B.L.E., D.G.G., D.T.S., K.A.H., M.A.S.M., P.G.M., S.L.S., S.M. and T.R.G. designed the project strategy and interpreted results. A.E.C., A.L.S., A.M.S., D.G.G., D.J.L., D.T.S., J.A.B., J.E.H., J.M.N., K.A.H., N.J.T., P.G.M., S.L.S., S.M., S.S., T.H., T.K. and Z.T. developed the assay. A.L.S., D.J.L., J.A.B., J.M.N., K.A.H., M. Duvet, N.J.T., P.A.C., P.G.M., S.C., S.S. and T.H. conducted small-molecule screening. A.L.S., A.R.K., B.M., C.H., D.J.L., D.P.S., F.A.-S., J.A.B., J.H.S., J.M.N., K.A.H., L.P.C., M. Dai, M. Duvet, M.J., N.J.T., P.G.M., R.N., R.P., S.C., S.S., T.H. and V.D. conducted follow-up studies. A.M.S., B.L.E., K.A.H., P.G.M. and T.R.G. drafted the paper.

Correspondence to Malcolm A S Moore or David T Scadden or Stuart L Schreiber or Benjamin L Ebert or Todd R Golub.

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Hartwell, K., Miller, P., Mukherjee, S. et al. Niche-based screening identifies small-molecule inhibitors of leukemia stem cells. Nat Chem Biol 9, 840–848 (2013) doi:10.1038/nchembio.1367

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