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Natural products reveal cancer cell dependence on oxysterol-binding proteins

Nature Chemical Biology volume 7, pages 639–647 (2011)Cite this article

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Abstract

Cephalostatin 1, OSW-1, ritterazine B and schweinfurthin A are natural products that potently, and in some cases selectively, inhibit the growth of cultured human cancer cell lines. The cellular targets of these small molecules have yet to be identified. We have discovered that these molecules target oxysterol binding protein (OSBP) and its closest paralog, OSBP-related protein 4L (ORP4L)—proteins not known to be involved in cancer cell survival. OSBP and the ORPs constitute an evolutionarily conserved protein superfamily, members of which have been implicated in signal transduction, lipid transport and lipid metabolism. The functions of OSBP and the ORPs, however, remain largely enigmatic. Based on our findings, we have named the aforementioned natural products ORPphilins. Here we used ORPphilins to reveal new cellular activities of OSBP. The ORPphilins are powerful probes of OSBP and ORP4L that will be useful in uncovering their cellular functions and their roles in human diseases.

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Figure 1: Chemical structures.
Figure 2: OSBP and ORP4L are high-affinity receptors of ORPphilins.
Figure 3: shRNA knockdown, protein expression and SAR experiments support OSBP and ORP4L as targets of ORPphilins.
Figure 4: Suppression of antiproliferative activity by 25-OHC administration and effects of lipoprotein-deficient media on proliferation.
Figure 5: ORPphilins induce OSBP translocation in cells.
Figure 6: Cephalostatin 1 and OSW-1 cause proteasome-dependent reduction of cellular OSBP concentrations.
Figure 7: ORPphilins block sphingomyelin biosynthesis.

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Acknowledgements

We thank the NCI for the generous gifts of schweinfurthin A. (J.A. Beutler) and ritterazine B. We gratefully acknowledge the aforementioned reagents provided by G. Romeo (Joslin Diabetes Center), M. Brown and J. Goldstein (University of Texas Southwestern Medical Center), H. Arai (University of Tokyo), B. Vogelstein (Johns Hopkins University) and D. Ory (Washington University, St. Louis). The beta-tubulin monoclonal antibody (E7) developed by M. Klymkowsky was obtained from the Developmental Studies Hybridoma Bank (National Institute of Child Health and Human Development, US National Institutes of Health (NIH) and University of Iowa, Department of Biology). R. King is acknowledged for helpful comments and discussions. T.B.P. would like to thank the Lundbeck foundation for a post-doctoral fellowship and the Danish Council for Independent Research–Natural Sciences for additional financial support. A.W.G.B gratefully thanks the Susan G. Komen for the Cure Foundation for providing a postdoctoral research fellowship. Financial support from the Novartis Institutes of Biomedical Research, the US NIH (grant no. R01GM090068), the Harvard Catalyst and The Harvard Clinical and Translational Science Center (NIH award no. UL1RR025758) is acknowledged. This work was conducted with support and financial contributions from Harvard University and its affiliated academic health care centers.

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  1. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA

    Anthony W G Burgett, Thomas B Poulsen, Kittikhun Wangkanont, D Ryan Anderson, Chikako Kikuchi, Kousei Shimada, Shuichi Okubo, Kevin C Fortner & Matthew D Shair

  2. School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan

    Yoshihiro Mimaki & Minpei Kuroda

  3. Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, USA

    Jason P Murphy, David J Schwalb, Eugene C Petrella, Ivan Cornella-Taracido, Markus Schirle & John A Tallarico

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Contributions

M.D.S., A.W.G.B., T.B.P., K.W. and D.R.A. conceived and designed the study. A.W.G.B., T.B.P., K.W. and D.R.A. implemented experiments. K.C.F. prepared cephalostatin 1. The OSBP and ORP4L molecular biology was done by K.W. and C.K. K.S. prepared OSW-1. S.O. performed early studies on p21 selectivity. Y.M. and M.K. supplied OSW-1 isolated from nature. A.W.G.B., D.R.A., M.S., J.P.M., E.C.P., D.J.S., I.C.-T. and J.A.T. performed the OSW-1 affinity chromatography experiments and subsequent quantitative mass spectrometry analysis. M.D.S., A.W.G.B., T.B.P. and K.W. wrote the paper.

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Correspondence to Matthew D Shair.

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Burgett, A., Poulsen, T., Wangkanont, K. et al. Natural products reveal cancer cell dependence on oxysterol-binding proteins. Nat Chem Biol 7, 639–647 (2011). https://doi.org/10.1038/nchembio.625

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