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Small-molecule activation of the TRAIL receptor DR5 in human cancer cells


Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) activates apoptosis through the death receptors DR4 and DR5. Because of its superior safety profile and high tumor specificity compared to other TNF family members, recombinant soluble TRAIL and agonistic antibodies against its receptors are actively being developed for clinical cancer therapy. Here, we describe the identification and characterization of the small molecules that directly target DR5 to initiate apoptosis in human cancer cells. The activity was initially discovered through a high-throughput chemical screen for compounds that promote cell death in synergy with a small-molecule mimetic of Smac, the antagonist for inhibitor of apoptosis protein. Structure-activity relationship studies yielded a more potent analog called bioymifi, which can act as a single agent to induce DR5 clustering and aggregation, leading to apoptosis. Thus, this study identified potential lead compounds for the development of small-molecule TRAIL mimics targeting DR5 for cancer therapy.

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Figure 1: Identification and chemical elucidation of the Smac synergists from the chemical library screen.
Figure 2: A functional analog of A2C2, bioymifi induces apoptosis.
Figure 3: Bioymifi-induced apoptosis occurs through a DR5-dependent extrinsic pathway.
Figure 4: Bioymifi promotes cell death independent of TRAIL.
Figure 5: Bioymifi selectively binds the ECD of DR5.
Figure 6: Bioymifi and A2C2 promote DR5 aggregation and activation.


  1. Green, D.R. Apoptotic pathways: ten minutes to dead. Cell 121, 671–674 (2005).

    Article  CAS  Google Scholar 

  2. Jiang, X. & Wang, X. Cytochrome c–mediated apoptosis. Annu. Rev. Biochem. 73, 87–106 (2004).

    Article  CAS  Google Scholar 

  3. Ashkenazi, A. & Dixit, V.M. Death receptors: signaling and modulation. Science 281, 1305–1308 (1998).

    Article  CAS  Google Scholar 

  4. Nagata, S. Apoptosis by death factor. Cell 88, 355–365 (1997).

    Article  CAS  Google Scholar 

  5. Wilson, N.S., Dixit, V. & Ashkenazi, A. Death receptor signal transducers: nodes of coordination in immune signaling networks. Nat. Immunol. 10, 348–355 (2009).

    Article  CAS  Google Scholar 

  6. Peter, M.E. & Krammer, P.H. The CD95(APO-1/Fas) DISC and beyond. Cell Death Differ. 10, 26–35 (2003).

    Article  CAS  Google Scholar 

  7. Sprick, M.R. et al. FADD/MORT1 and caspase-8 are recruited to TRAIL receptors 1 and 2 and are essential for apoptosis mediated by TRAIL receptor 2. Immunity 12, 599–609 (2000).

    Article  CAS  Google Scholar 

  8. Kischkel, F.C. et al. Apo2L/TRAIL-dependent recruitment of endogenous FADD and caspase-8 to death receptors 4 and 5. Immunity 12, 611–620 (2000).

    Article  CAS  Google Scholar 

  9. Varfolomeev, E. et al. Molecular determinants of kinase pathway activation by Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand. J. Biol. Chem. 280, 40599–40608 (2005).

    Article  CAS  Google Scholar 

  10. Crook, N.E., Clem, R.J. & Miller, L.K. An apoptosis-inhibiting baculovirus gene with a zinc finger–like motif. J. Virol. 67, 2168–2174 (1993).

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Deveraux, Q.L. et al. IAPs block apoptotic events induced by caspase-8 and cytochrome c by direct inhibition of distinct caspases. EMBO J. 17, 2215–2223 (1998).

    Article  CAS  Google Scholar 

  12. Roy, N., Deveraux, Q.L., Takahashi, R., Salvesen, G.S. & Reed, J.C. The c-IAP-1 and c-IAP-2 proteins are direct inhibitors of specific caspases. EMBO J. 16, 6914–6925 (1997).

    Article  CAS  Google Scholar 

  13. Li, L. et al. A small molecule Smac mimic potentiates TRAIL- and TNFα-mediated cell death. Science 305, 1471–1474 (2004).

    Article  CAS  Google Scholar 

  14. Wang, L., Du, F. & Wang, X. TNF-α induces two distinct caspase-8 activation pathways. Cell 133, 693–703 (2008).

    Article  CAS  Google Scholar 

  15. Petersen, S.L. et al. Autocrine TNFα signaling renders human cancer cells susceptible to Smac-mimetic–induced apoptosis. Cancer Cell 12, 445–456 (2007).

    Article  CAS  Google Scholar 

  16. Hanahan, D. & Weinberg, R.A. The hallmarks of cancer. Cell 100, 57–70 (2000).

    Article  CAS  Google Scholar 

  17. Danial, N.N. & Korsmeyer, S.J. Cell death: critical control points. Cell 116, 205–219 (2004).

    Article  CAS  Google Scholar 

  18. Vince, J.E. et al. IAP antagonists target cIAP1 to induce TNFα-dependent apoptosis. Cell 131, 682–693 (2007).

    Article  CAS  Google Scholar 

  19. Wiezorek, J., Holland, P. & Graves, J. Death receptor agonists as a targeted therapy for cancer. Clin. Cancer Res. 16, 1701–1708 (2010).

    Article  CAS  Google Scholar 

  20. Ashkenazi, A., Holland, P. & Eckhardt, S.G. Ligand-based targeting of apoptosis in cancer: the potential of recombinant human apoptosis ligand 2/tumor necrosis factor-related apoptosis-inducing ligand (rhApo2L/TRAIL). J. Clin. Oncol. 26, 3621–3630 (2008).

    Article  CAS  Google Scholar 

  21. Feoktistova, M. et al. cIAPs block ripoptosome formation, a RIP1/caspase-8 containing intracellular cell death complex differentially regulated by cFLIP isoforms. Mol. Cell 43, 449–463 (2011).

    Article  CAS  Google Scholar 

  22. Alberti, S., Halfmann, R., King, O., Kapila, A. & Lindquist, S. A systematic survey identifies prions and illuminates sequence features of prionogenic proteins. Cell 137, 146–158 (2009).

    Article  CAS  Google Scholar 

  23. Hou, F. et al. MAVS forms functional prion-like aggregates to activate and propagate antiviral innate immune response. Cell 146, 448–461 (2011).

    Article  CAS  Google Scholar 

  24. Jin, Z. et al. Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cell 137, 721–735 (2009).

    Article  CAS  Google Scholar 

  25. Siegel, R.M. et al. SPOTS: signaling protein oligomeric transduction structures are early mediators of death receptor-induced apoptosis at the plasma membrane. J. Cell Biol. 167, 735–744 (2004).

    Article  CAS  Google Scholar 

  26. Wagner, K.W. et al. Death-receptor O-glycosylation controls tumor-cell sensitivity to the proapoptotic ligand Apo2L/TRAIL. Nat. Med. 13, 1070–1077 (2007).

    Article  CAS  Google Scholar 

  27. Fulda, S., Wick, W., Weller, M. & Debatin, K.M. Smac agonists sensitize for Apo2L/TRAIL- or anticancer drug–induced apoptosis and induce regression of malignant glioma in vivo. Nat. Med. 8, 808–815 (2002).

    Article  CAS  Google Scholar 

  28. Chen, G. & Goeddel, D.V. TNF-R1 signaling: a beautiful pathway. Science 296, 1634–1635 (2002).

    Article  CAS  Google Scholar 

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We thank M. Zhao for NMR and HPLC/MS analysis; S.L. Petersen for reagents and discussions; L. Zhao for the toxicological experiments; S.L. McKnight for helpful suggestions; M. Roth for proposing the chemical library screen; D. Frantz, J. Ready and A. Wang for synthesis of initial hits; Y. Liu, Z. Zhang, Q. Liu and L. Shang for comments of the manuscript; S.-C. Tso and T. Scheuermann for detecting the interaction between DR5 and bioymifi; and K. Phelps and A. Budge for the cell imaging. This work is supported by a program project grant from the US National Cancer Institute (2P01 CA095471-06 to Xiaodong Wang and G.W.) and the National Natural Science Foundation of China (21072150 and 21222209 to X.L.).

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Authors and Affiliations



Xiaodong Wang, X.L. and G.W. designed the study; G.W., H.Y., N.W., D.L.H., R.H., L.L. and L.W. performed and analyzed the biological experiments; Xiaoming Wang, J.N., P.H. and X.L. performed all of the chemical syntheses; S.C. performed the MS analysis; S.W. implemented the chemical library screen; and Xiaodong Wang, X.L. and G.W. wrote the manuscript.

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Correspondence to Gelin Wang, Xiaoguang Lei or Xiaodong Wang.

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The authors declare no competing financial interests.

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Wang, G., Wang, X., Yu, H. et al. Small-molecule activation of the TRAIL receptor DR5 in human cancer cells. Nat Chem Biol 9, 84–89 (2013).

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