Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Induction of IL6/STAT3-dependent TRAIL expression that contributes to the therapeutic efficacy of osimertinib in EGFR mutant NSCLC cells

Oncogene volume 44, pages 2315–2327 (2025)Cite this article

Subjects

Abstract

The third-generation mutation-selective EGFR tyrosine kinase inhibitor (EGFR-TKI) osimertinib (or AZD9291) effectively induces apoptosis in EGFR mutant (EGFRm) non-small cell lung cancer (NCSLC) cells. However, the underlying mechanisms have not been fully elucidated. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL or TNFSF10) is known as a death ligand that initiates apoptosis via binding to its cell surface death receptors such as DR5. In this study, we found that osimertinib and other EGFR-TKIs increased the expression of TRAIL primarily in EGFRm NSCLC cell lines. This effect was accompanied with increased IL6 expression and STAT3 activation. Inhibition of STAT3 with either protein degradation or gene knockout abrogated the ability of osimertinib or recombinant human IL6 to elevate TRAIL levels. Moreover, osimertinib increased STAT3-dependent transcription of TRAIL via two STAT3 novel binding sites present in the TRAIL 5’flanking region. Hence, osimertinib induces IL6/STAT3-mediated TRAIL expression in EGFRm NSCLC cells. While osimertinib lost the ability to induce TRAIL expression in osimertinib-resistant EGFRm NSCLC, knockdown or knockout of TRAIL in sensitive EGFRm NSCLC cells rendered them less sensitive to osimertinib both in vitro and in vivo. Thus, TRAIL elevation contributes to the induction of apoptosis by osimertinib in EGFRm NSCLC cells. Furthermore, osimertinib increased membrane-bound TRAIL and DR5 membrane clustering and DR5 knockdown significantly compromised the cell-killing effect of osimertinib, together suggesting a DR5-dependent effect. Collectively, this study has revealed a previously undiscovered connection between TRAIL induction and osimertinib-induced apoptosis in EGFRm NSCLC cells, increasing our understanding of mechanisms accounting for apoptosis induced by osimertinib.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Osimertinib and other EGFR-TKIs induce TRAIL gene expression in EGFRm NSCLC cell lines and tumors through increasing gene transcription.
Fig. 2: Osimertinib activates STAT3 together with induction of IL6 expression and induces IL6/STAT3 dependent TRAIL expression in EGFRm NSCLC cells.
Fig. 3: Osimertinib increases TRAIL gene transactivation through non-canonical STAT3 binding sites present in 5’-flanking region of the TRAIL gene.
Fig. 4: Osimertinib-resistant cell lines are not responsive to activation of STAT3 and induction of TRAIL by osimertinib, which contributes to decreased survival and enhanced apoptosis by osimertinib.
Fig. 5: TRAIL knockout in PC-9 cells compromises the effects of osimertinib on decreasing cell survival, inducing apoptosis, and suppressing tumor growth in vivo.
Fig. 6: Osimertinib decreases cell survival and induces apoptosis in EGFRm NSCLC cell lines in part through a DR5-dependent mechanism albeit independent of soluble TRAIL.
Fig. 7: Osimertinib enhances TRAIL and DR5 interaction, induces DR5 clustering, and increases membrane-bound TRAIL levels, suggesting a role of TRAIL induction and subsequent DR5 cluster formation in mediating apoptosis induced by osimertinib or other EGFR-TKIs.

Similar content being viewed by others

Data availability

All data generated in this study are presented in the article.

References

  1. Tartarone A, Lerose R. Clinical approaches to treat patients with non-small cell lung cancer and epidermal growth factor receptor tyrosine kinase inhibitor acquired resistance. Ther Adv Respir Dis. 2015;9:242–50.

    Article  CAS  PubMed  Google Scholar 

  2. Nagasaka M, Zhu VW, Lim SM, Greco M, Wu F, Ou SI. Beyond osimertinib: the development of third-generation egfr tyrosine kinase inhibitors for advanced EGFR+ NSCLC. J Thorac Oncol. 2021;16:740–63.

    Article  CAS  PubMed  Google Scholar 

  3. Ramalingam SS, Vansteenkiste J, Planchard D, Cho BC, Gray JE, Ohe Y, et al. Overall Survival with Osimertinib in Untreated, EGFR-Mutated Advanced NSCLC. N Engl J Med. 2020;382:41–50.

    Article  CAS  PubMed  Google Scholar 

  4. Blaquier JB, Ortiz-Cuaran S, Ricciuti B, Mezquita L, Cardona AF, Recondo G. Tackling osimertinib resistance in EGFR mutant non-small cell lung cancer. Clin Cancer Res. 2023;29:3579–91.

  5. Zalaquett Z, Catherine Rita Hachem M, Kassis Y, Hachem S, Eid R, Raphael Kourie H, et al. Acquired resistance mechanisms to osimertinib: the constant battle. Cancer Treat Rev. 2023;116:102557.

    Article  CAS  PubMed  Google Scholar 

  6. Gomatou G, Syrigos N, Kotteas E. Osimertinib resistance: molecular mechanisms and emerging treatment options. Cancers. 2023;15:841.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Tang ZH, Lu JJ. Osimertinib resistance in non-small cell lung cancer: mechanisms and therapeutic strategies. Cancer Lett. 2018;420:242–6.

    Article  CAS  PubMed  Google Scholar 

  8. Hengartner MO. The biochemistry of apoptosis. Nature. 2000;407:770–6.

    Article  CAS  PubMed  Google Scholar 

  9. Fesik SW. Promoting apoptosis as a strategy for cancer drug discovery. Nat Rev Cancer. 2005;5:876–85.

    Article  CAS  PubMed  Google Scholar 

  10. Shi P, Oh YT, Deng L, Zhang G, Qian G, Zhang S, et al. Overcoming acquired resistance to AZD9291, a third-generation EGFR inhibitor, through modulation of MEK/ERK-dependent Bim and Mcl-1 degradation. Clin Cancer Res. 2017;23:6567–79.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Shi P, Zhang S, Zhu L, Qian G, Ren H, Ramalingam SS, et al. The third-generation EGFR inhibitor, osimertinib, promotes c-FLIP degradation, enhancing apoptosis including TRAIL-induced apoptosis in NSCLC cells with activating EGFR mutations. Transl Oncol. 2019;12:705–13.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Wiley SR, Schooley K, Smolak PJ, Din WS, Huang CP, Nicholl JK, et al. Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity. 1995;3:673–82.

    Article  CAS  PubMed  Google Scholar 

  13. Montinaro A, Walczak H. Harnessing TRAIL-induced cell death for cancer therapy: a long walk with thrilling discoveries. Cell Death Differ. 2023;30:237–49.

    Article  CAS  PubMed  Google Scholar 

  14. Pimentel JM, Zhou JY, Wu GS. The role of TRAIL in apoptosis and immunosurveillance in cancer. Cancers. 2023;15:2752.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Smyth MJ, Takeda K, Hayakawa Y, Peschon JJ, van den Brink MR, Yagita H. Nature’s TRAIL-on a path to cancer immunotherapy. Immunity. 2003;18:1–6.

    Article  CAS  PubMed  Google Scholar 

  16. Falschlehner C, Schaefer U, Walczak H. Following TRAIL’s path in the immune system. Immunology. 2009;127:145–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Liu S, Polsdofer EV, Zhou L, Ruan S, Lyu H, Hou D, et al. Upregulation of endogenous TRAIL-elicited apoptosis is essential for metformin-mediated antitumor activity against TNBC and NSCLC. Mol Ther Oncolytics. 2021;21:303–14.

    Article  PubMed  PubMed Central  Google Scholar 

  18. Allen JE, Krigsfeld G, Mayes PA, Patel L, Dicker DT, Patel AS, et al. Dual inactivation of Akt and ERK by TIC10 signals Foxo3a nuclear translocation, TRAIL gene induction, and potent antitumor effects. Sci Transl Med. 2013;5:171ra117.

    Article  Google Scholar 

  19. Lund P, Kotova I, Kedinger V, Khanwalkar H, Voltz E, Hahn WC, et al. Transformation-dependent silencing of tumor-selective apoptosis-inducing TRAIL by DNA hypermethylation is antagonized by decitabine. Mol Cancer Ther.2011;10:1611–23.

    Article  CAS  PubMed  Google Scholar 

  20. Cho SD, Yoon K, Chintharlapalli S, Abdelrahim M, Lei P, Hamilton S, et al. Nur77 agonists induce proapoptotic genes and responses in colon cancer cells through nuclear receptor-dependent and nuclear receptor-independent pathways. Cancer Res. 2007;67:674–83.

    Article  CAS  PubMed  Google Scholar 

  21. Oh YT, Yue P, Zhou W, Balko JM, Black EP, Owonikoko TK, et al. Oncogenic Ras and B-Raf proteins positively regulate death receptor 5 expression through co-activation of ERK and JNK signaling. J Biol Chem. 2012;287:257–67.

    Article  CAS  PubMed  Google Scholar 

  22. Oh YT, Liu X, Yue P, Kang S, Chen J, Taunton J, et al. ERK/ribosomal S6 kinase (RSK) signaling positively regulates death receptor 5 expression through co-activation of CHOP and Elk1. J Biol Chem. 2010;285:41310–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Shi P, Oh YT, Zhang G, Yao W, Yue P, Li Y, et al. Met gene amplification and protein hyperactivation is a mechanism of resistance to both first and third-generation EGFR inhibitors in lung cancer treatment. Cancer Lett. 2016;380:494–504.

    Article  CAS  PubMed  Google Scholar 

  24. Xu J, Zhou JY, Wu GS. Tumor necrosis factor-related apoptosis-inducing ligand is required for tumor necrosis factor alpha-mediated sensitization of human breast cancer cells to chemotherapy. Cancer Res. 2006;66:10092–9.

    Article  CAS  PubMed  Google Scholar 

  25. Maadi H, Nami B, Wang Z. Dimerization assessment of epithelial growth factor family of receptor tyrosine kinases by using cross-linking reagent. Methods Mol Biol. 2017;1652:101–8.

    Article  CAS  PubMed  Google Scholar 

  26. Turk HF, Chapkin RS. Analysis of epidermal growth factor receptor dimerization by BS(3) cross-linking. Methods Mol Biol. 2015;1233:25–34.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Zhang S, Qian G, Shi P, Zhang G, Fan S, He Y, et al. Downregulation of death receptor 4 is tightly associated with positive response of EGFR mutant lung cancer to EGFR-targeted therapy and improved prognosis. Theranostics. 2021;11:3964–80.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Zhu L, Chen Z, Zang H, Fan S, Gu J, Zhang G, et al. Targeting c-Myc to overcome acquired resistance of EGFR mutant NSCLC cells to the third-generation EGFR tyrosine kinase inhibitor, osimertinib. Cancer Res. 2021;81:4822–34.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Brown K, Comisar C, Witjes H, Maringwa J, de Greef R, Vishwanathan K, et al. Population pharmacokinetics and exposure-response of osimertinib in patients with non-small cell lung cancer. Br J Clin Pharm. 2017;83:1216–26.

    Article  CAS  Google Scholar 

  30. Matsuda T. The physiological and pathophysiological role of IL-6/STAT3-mediated signal transduction and STAT3 binding partners in therapeutic applications. Biol Pharm Bull. 2023;46:364–78.

    Article  CAS  PubMed  Google Scholar 

  31. Bai L, Zhou H, Xu R, Zhao Y, Chinnaswamy K, McEachern D, et al. A potent and selective small-molecule degrader of STAT3 achieves complete tumor regression in vivo. Cancer Cell. 2019;36:498–511.e417.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Zhou H, Bai L, Xu R, Zhao Y, Chen J, McEachern D, et al. Structure-based discovery of SD-36 as a potent, selective, and efficacious PROTAC degrader of STAT3 protein. J Med Chem. 2019;62:11280–300.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Wang J, Wang Y, Zheng C, Hou K, Zhang T, Qu X, et al. Tyrosine kinase inhibitor-induced IL-6/STAT3 activation decreases sensitivity of EGFR-mutant non-small cell lung cancer to icotinib. Cell Biol Int. 2018;42:1292–9.

    Article  CAS  PubMed  Google Scholar 

  34. Ishiguro Y, Ishiguro H, Miyamoto H. Epidermal growth factor receptor tyrosine kinase inhibition up-regulates interleukin-6 in cancer cells and induces subsequent development of interstitial pneumonia. Oncotarget. 2013;4:550–9.

    Article  PubMed  PubMed Central  Google Scholar 

  35. Li L, Li Z, Lu C, Li J, Zhang K, Lin C, et al. Ibrutinib reverses IL-6-induced osimertinib resistance through inhibition of Laminin alpha5/FAK signaling. Commun Biol. 2022;5:155.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Vallania F, Schiavone D, Dewilde S, Pupo E, Garbay S, Calogero R, et al. Genome-wide discovery of functional transcription factor binding sites by comparative genomics: the case of Stat3. Proc Natl Acad Sci USA. 2009;106:5117–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Seidel HM, Milocco LH, Lamb P, Darnell JE Jr, Stein RB, Rosen J. Spacing of palindromic half sites as a determinant of selective STAT (signal transducers and activators of transcription) DNA binding and transcriptional activity. Proc Natl Acad Sci USA. 1995;92:3041–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Ihle JN. STATs: signal transducers and activators of transcription. Cell. 1996;84:331–4.

    Article  CAS  PubMed  Google Scholar 

  39. von Karstedt S, Montinaro A, Walczak H. Exploring the TRAILs less travelled: TRAIL in cancer biology and therapy. Nat Rev Cancer. 2017;17:352–66.

    Article  Google Scholar 

  40. Pan L, Fu TM, Zhao W, Zhao L, Chen W, Qiu C, et al. Higher-order clustering of the transmembrane anchor of DR5 drives signaling. Cell. 2019;176:1477–89.e1414.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Du G, Zhao L, Zheng Y, Belfetmi A, Cai T, Xu B, et al. Autoinhibitory structure of preligand association state implicates a new strategy to attain effective DR5 receptor activation. Cell Res. 2023;33:131–46.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Lee HK, Noh MH, Hong SW, Kim SM, Kim SH, Kim YS, et al. Erlotinib activates different cell death pathways in EGFR-mutant lung cancer cells grown in 3D versus 2D culture systems. Anticancer Res. 2021;41:1261–9.

    Article  CAS  PubMed  Google Scholar 

  43. Chen S, Fu L, Raja SM, Yue P, Khuri FR, Sun SY. Dissecting the roles of DR4, DR5 and c-FLIP in the regulation of geranylgeranyltransferase I inhibition-mediated augmentation of TRAIL-induced apoptosis. Mol Cancer. 2010;9:23.

    Article  PubMed  PubMed Central  Google Scholar 

  44. Naval J, de Miguel D, Gallego-Lleyda A, Anel A, Martinez-Lostao L. Importance of TRAIL Molecular anatomy in receptor oligomerization and signaling. Implications for cancer therapy. Cancers. 2019;11:444.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Chang Q, Bournazou E, Sansone P, Berishaj M, Gao SP, Daly L, et al. The IL-6/JAK/Stat3 feed-forward loop drives tumorigenesis and metastasis. Neoplasia. 2013;15:848–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We are thankful to Dr. Gen-Sheng Wu for providing reporter constructs with TRAIL 5’-flanking region used in this study. We are grateful to Dr. Anthea Hammond in our department for editing the manuscript. This work was supported by NIH/NCI R01 CA223220 (to SYS) and UG1 CA233259 (to SSR/SY) and by the David A. Cole Family Professorship. SSR and SYS are Georgia Research Alliance Distinguished Cancer Research Scholars. SYS is a David A. Cole Family Professor.

Author information

Authors and Affiliations

  1. Department of Hematology and Medical Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA, USA

    You-Take Oh, Zhen Chen, Dongsheng Wang, Suresh S. Ramalingam & Shi-Yong Sun

Authors
  1. You-Take Oh
    View author publications

    Search author on:PubMed Google Scholar

  2. Zhen Chen
    View author publications

    Search author on:PubMed Google Scholar

  3. Dongsheng Wang
    View author publications

    Search author on:PubMed Google Scholar

  4. Suresh S. Ramalingam
    View author publications

    Search author on:PubMed Google Scholar

  5. Shi-Yong Sun
    View author publications

    Search author on:PubMed Google Scholar

Contributions

Conceptualization: Y-TO, and S-YS. Investigation: Y-TO, ZC, and DW. Methodology: Y-TO, ZC, and DW. Writing/Original Draft: Y-TO, and S-YS. Review/Editing: ZC, DW, and SSR. Funding Acquisition: SSR, and S-YS. Supervision: S-Y.S.

Corresponding author

Correspondence to Shi-Yong Sun.

Ethics declarations

Competing interests

SSR is on consulting/advisory board for AstraZeneca, BMS, Merck, Roche, Tesaro and Amgen. Other authors disclose that they have no potential conflicts of interest.

Ethics approval and consent to participate

Animal experiment was approved by the Institutional Animal Care and Use Committee (IACUC) of Emory University (PROTO201700718). All methods were performed in accordance with the relevant guidelines and regulations.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Oh, YT., Chen, Z., Wang, D. et al. Induction of IL6/STAT3-dependent TRAIL expression that contributes to the therapeutic efficacy of osimertinib in EGFR mutant NSCLC cells. Oncogene 44, 2315–2327 (2025). https://doi.org/10.1038/s41388-025-03381-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41388-025-03381-5

Search

Advanced search

Quick links