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Kinome screening for regulators of the estrogen receptor identifies LMTK3 as a new therapeutic target in breast cancer

Nature Medicine volume 17, pages 715–719 (2011)Cite this article

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Abstract

Therapies targeting estrogen receptor α (ERα, encoded by ESR1) have transformed the treatment of breast cancer. However, large numbers of women relapse, highlighting the need for the discovery of new regulatory targets modulating ERα pathways1,2,3,4,5. An siRNA screen identified kinases whose silencing alters the estrogen response including those previously implicated in regulating ERα activity (such as mitogen-activated protein kinase and AKT). Among the most potent regulators was lemur tyrosine kinase-3 (LMTK3), for which a role has not previously been assigned. In contrast to other modulators of ERα activity, LMTK3 seems to have been subject to Darwinian positive selection, a noteworthy result given the unique susceptibility of humans to ERα+ breast cancer. LMTK3 acts by decreasing the activity of protein kinase C (PKC) and the phosphorylation of AKT (Ser473), thereby increasing binding of forkhead box O3 (FOXO3) to the ESR1 promoter. LMTK3 phosphorylated ERα, protecting it from proteasomal degradation in vitro. Silencing of LMTK3 reduced tumor volume in an orthotopic mouse model and abrogated proliferation of ERα+ but not ERα− cells, indicative of its role in ERα activity. In human cancers, LMTK3 abundance and intronic polymorphisms were significantly associated with disease-free and overall survival and predicted response to endocrine therapies. These findings yield insights into the natural history of breast cancer in humans and reveal LMTK3 as a new therapeutic target.

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Figure 1: High-throughput siRNA screen identifies kinases modulating ERα transcriptional activity.
Figure 2: Mechanisms of LMTK3 action on ERα transcription and translation.
Figure 3: Association of LMTK3 expression and germline polymorphisms with clinical outcome.
Figure 4: Tumor growth inhibition by in vivo LMTK3 siRNA in an orthotopic mouse model.

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Acknowledgements

We would like to thank P. Hexley and J. Elliott of the Flow Cytometry Core Facility (Imperial College London) for analyzing FACS data, J. Gronau for helping with confocal microscopy, P. Trivedi for performing the immunohistochemistry staining and T. Rapoz D'Silva for helping in the laboratory. We are grateful to G. Dranitsaris (Princess Margaret Hospital, Toronto) for statistical advice. Li93 software was a kind gift from W. Hsiung-Li (University of Chicago). The single-nucleotide polymorphism analyses were funded in part by US National Institutes of Health grant 5 P30CA14089-27I. S.A. and R.C.C. are supported by grants from Cancer Research UK and the Department of Health–funded Imperial College Cancer Medicine Centre (ECMC) grant. We thank R. and E. Girling and friends for their support.

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Authors and Affiliations

  1. Department of Surgery and Cancer, Imperial College London, Hammersmith Hospital Campus, London, UK

    Georgios Giamas, Aleksandra Filipović, Jimmy Jacob, Hua Zhang, Belul Assefa Shifa, Andrew Photiou, Leandro Castellano, R Charles Coombes, Simak Ali & Justin Stebbing

  2. Evolutionary Genomics, Inc., Lafayette, Colorado, USA

    Walter Messier

  3. Division of Medical Oncology, University of Southern California, Norris Comprehensive Cancer Center, Keck School of Medicine, Los Angeles, California, USA

    Dongyun Yang, Wu Zhang & Heinz-Josef Lenz

  4. Drug Discovery Centre, Imperial College London, Biochemistry Building, South Kensington Campus, London, UK.,

    Cathy Tralau-Stewart

  5. Histopathology, School of Molecular Medical Sciences, University of Nottingham and Nottingham University NHS Hospital Trust, Nottingham, UK

    Andrew R Green & Ian O Ellis

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  1. Georgios Giamas
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Contributions

G.G. and J.S. conceived of the study, initiated, designed, supervised and conducted most of the experiments and wrote the manuscript. W.M., S.A., H.-J.L., C.T.-S. and R.C.C. contributed to manuscript editing. G.G. and J.J. performed the kinome screening. A.F., B.A.S., A.P., L.C. and H.Z. performed in vitro experiments (including proliferation assays, quantitative RT-PCR and FACS). A.F. and J.S. performed the immunohistochemistry scoring. D.Y., W.Z. and H.J.L. generated the single-nucleotide polymorphism data. W.M. produced all the evolutionary data. A.R.G. and I.O.E. performed the statistical analysis of the clinical data. All authors discussed the results, conceived further experiments, commented on the manuscript and approved the final submitted version.

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Correspondence to Georgios Giamas or Justin Stebbing.

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The authors declare no competing financial interests.

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Giamas, G., Filipović, A., Jacob, J. et al. Kinome screening for regulators of the estrogen receptor identifies LMTK3 as a new therapeutic target in breast cancer. Nat Med 17, 715–719 (2011). https://doi.org/10.1038/nm.2351

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