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Active Pin1 is a key target of all-trans retinoic acid in acute promyelocytic leukemia and breast cancer

Abstract

A common key regulator of oncogenic signaling pathways in multiple tumor types is the unique isomerase Pin1. However, available Pin1 inhibitors lack the required specificity and potency for inhibiting Pin1 function in vivo. By using mechanism-based screening, here we find that all-trans retinoic acid (ATRA)—a therapy for acute promyelocytic leukemia (APL) that is considered the first example of targeted therapy in cancer, but whose drug target remains elusive—inhibits and degrades active Pin1 selectively in cancer cells by directly binding to the substrate phosphate- and proline-binding pockets in the Pin1 active site. ATRA-induced Pin1 ablation degrades the protein encoded by the fusion oncogene PML–RARA and treats APL in APL cell and animal models as well as in human patients. ATRA-induced Pin1 ablation also potently inhibits triple-negative breast cancer cell growth in human cells and in animal models by acting on many Pin1 substrate oncogenes and tumor suppressors. Thus, ATRA simultaneously blocks multiple Pin1-regulated cancer-driving pathways, an attractive property for treating aggressive and drug-resistant tumors.

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Figure 1: Mechanism-based screening identifies ATRA as a submicromolar Pin1 inhibitor that binds to the Pin1 active site.
Figure 2: ATRA causes Pin1 degradation and inhibits its oncogenic function in cells.
Figure 3: Pin1 is a critical target for ATRA to induce PML–RAR-α degradation and inhibit proliferation in APL cells.
Figure 4: Inhibition of Pin1 by ATRA or other compounds causes PML–RAR-α degradation and treats APL in cell and mouse models and human subjects.
Figure 5: ATRA ablates active Pin1 and thereby turns off oncogenes and turns on tumor suppressors in breast cancer.
Figure 6: ATRA exerts potent anticancer activity against TNBC in vivo by ablating Pin1 and thereby blocking multiple cancer pathways simultaneously.

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Acknowledgements

We thank W.G. Kaelin Jr., N. Gray, J. Clardy and A. Chakraborty for constructive advice; and H. de Thé (INSERM) for RAR-α, RAR-β, and RAR-γ triple-KO MEFs originally generated by P.A. Chambon (Université de Strasbourg); C. Ng for assistance with immunostaining and T. Garvey for editing the manuscript. S.W. is a recipient of a Susan G. Komen for the Cure postdoctoral fellowship (KG111233). The work is supported by grants from the US National Institutes of Health (R01CA167677, R03DA031663 and R01HL111430 to K.P.L.).

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S.W. designed the studies, performed the experiments, interpreted the data, and wrote the manuscript; S.K. helped characterize ATRA binding to and inhibition of Pin1; L.K., J.G. and M.R. helped design and conduct APL-related experiments; W.L. and Y.Z. determined the Pin1–ATRA co-crystal structure; M.N., M.L., Y.Y., A.K., H.H., and C.H.C. provided various technical assistances; M.-H.Y and T.H.L. performed Pin1 and DAPK1 immunostaining; G.B. and H.W. helped analyze Pin1 and ATRA binding; N.J.M. and S.C. provided advice on the FP-HTS screen; E.M.R. and F.L.-C. provided human APL samples; L.C.C. advised the project; P.P.P. advised the project, interpreted the data and reviewed the manuscript; X.Z.Z. developed the original Pin1 FP-HTS and worked with S.W. to identify ATRA; X.Z.Z. and K.P.L. conceived and supervised the project, designed the studies, interpreted the data, and wrote the manuscript.

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Correspondence to Xiao Zhen Zhou or Kun Ping Lu.

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Competing interests

K.P.L. and X.Z.Z. are the inventors of Pin1 inhibition technology, which was licensed by BIDMC to Pinteon Therapeutics. K.P.L. and X.Z.Z. own equity in and consult for Pinteon. K.P.L. also serves on its board of directors. Their interests were reviewed and are managed by BIDMC in accordance with its conflict of interest policy.

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Wei, S., Kozono, S., Kats, L. et al. Active Pin1 is a key target of all-trans retinoic acid in acute promyelocytic leukemia and breast cancer. Nat Med 21, 457–466 (2015). https://doi.org/10.1038/nm.3839

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