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TFF3 drives Hippo dependent EGFR-TKI resistance in lung adenocarcinoma

Abstract

Intrinsic and acquired resistance represent major obstacles to optimize outcomes in epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) targeted therapy in lung adenocarcinoma (LUAD). Hence, a deeper understanding of EGFR-TKI resistance mechanisms in LUAD will potentially assist in formulating strategies to delay or overcome such resistance. Herein, it was observed that trefoil factor 3 (TFF3) is a crucial mediator of the LUAD EGFR-TKI response. TFF3 conferred intrinsic resistance to EGFR inhibition in LUAD by promotion of EGFR activation. TFF3 expression was also increased in acquired EGFR-TKI resistant LUAD, accompanied by reduced EGFR activation. YAP, a key mediator of the Hippo signaling, was positively regulated by TFF3 by post-transcriptional mechanisms and was responsible for acquired EGFR-TKI resistance mediated by TFF3. Inhibition of TFF3 by a small molecule inhibitor not only enhanced EGFR-TKI sensitivity in LUAD cells but also restored the sensitivity of acquired EGFR-TKI resistant LUAD cells to EGFR-TKIs in vitro and in vivo. These findings demonstrate a pivotal function of TFF3 in mediating both intrinsic and acquired EGFR-TKI resistance in LUAD and may offer a potential therapeutic mechanism for delaying or overcoming resistance to EGFR-TKIs.

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Fig. 1: High TFF3 expression correlates with worse prognosis in LUAD patients and forced expression of TFF3 reduces EGFR-TKI sensitivity in LUAD cells via EGFR signaling.

Fig. 2: Depletion of TFF3 or inhibition of TFF3 with AMPC increases the sensitivity of LUAD to EGFR-TKIs.

Fig. 3: TFF3 expression is elevated in acquired EGFR-TKI resistant LUAD.

Fig. 4: Depletion of TFF3 or inhibition of TFF3 with AMPC restores the sensitivity of acquired EGFR-TKI resistant LUAD cells to EGFR-TKIs.

Fig. 5: Inhibition of TFF3 with AMPC decreases cancer cell proliferation, promotes apoptosis and restores EGFR-TKI sensitivity in acquired EGFR-TKI resistant LUAD xenografts.

Fig. 6: Inactivation of Hippo signaling pathway is required for acquired EGFR-TKI resistance.

Fig. 7: TFF3 inactivates Hippo signaling in acquired EGFR-TKI resistance.

Data availability

The data sets used in this study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors would like to thank Dr. Xiaoming Huang and Dr. Hui Guo for their valuable technical support. This work was supported by National Natural Science Foundation of China (82172618 and 82102768); the Shenzhen Key Laboratory of Innovative Oncotherapeutics (ZDSYS20200820165400003) (Shenzhen Science and Technology Innovation Commission), China; Shenzhen Development and Reform Commission Subject Construction Project ([2017] 1434), China; Universities Stable Funding Key Projects (WDZC20200821150704001), China; Guangdong Basic and Applied Basic Research Foundation (2020A1515111064), China; the Shenzhen Bay Laboratory, Oncotherapeutics (21310031), China; Overseas Research Cooperation Project (HW2020008) (Tsinghua Shenzhen International Graduate School), China; TBSI Faculty Start-Up Funds, China and Start-up Research Fund, BNU-HKBU United International College (UICR0700101-24), China.

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Authors and Affiliations

Institute of Biopharmaceutical and Health Engineering, Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen, 518055, PR China

Shuwei Zhang, Yan Qin Tan, Vijay Pandey & Peter E. Lobie

Guangdong Provincial Key Laboratory IRADS and Food Science and Technology Program, Department of Life Sciences, BNU-HKBU United International College, Zhuhai, 519087, Guangdong, PR China

Yan Qin Tan

Shenzhen Bay Laboratory, Shenzhen, 518055, Guangdong, PR China

Xi Zhang, Tao Zhu & Peter E. Lobie

Labortory of Chemical Biology, Department of Studies in Organic Chemistry, University of Mysore, Manasagangotri, Mysore, 570005, India

Basappa Basappa

Department of Oncology, The First Affiliated Hospital of USTC, Center for Advanced Interdisciplinary Science and Biomedicine of IHM, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230027, PR China

Tao Zhu

Key Laboratory of Immune Response and Immunotherapy, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230027, PR China

Tao Zhu

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Contributions

P.E.L., V.P. and S.Z. designed the study; Basappa synthesized AMPC; S.Z. and Y.Q.T. performed the experiments; S.Z., Y.Q.T., T.Z. and P.E.L. analyzed the data; X.Z. provided resources and cell lines; S.Z. wrote the draft; P.E.L. and Y.Q.T. revised the paper. All authors read and approved the final manuscript.

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Correspondence to Vijay Pandey or Peter E. Lobie.

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Competing interests

The authors declare the following competing financial interest(s): P.E.L. and T.Z. have previously consulted for Perseis Therapeutics Ltd. P.E.L. is named on PCT application numbers WO 2006/69253 and WO 2008/042435 and US provisional application number 61/059558 and derivatives thereof. V.P., B.B., and P.E.L. are named as inventors on PCT application WO/2018/226155 (PCT/SG2018/050277), Compounds, As Inhibitors of TFF3 Dimerization, Methods and Applications Thereof (and derivatives thereof including US Patent 11,141,402). P.E.L. is an equity holder in Sinotar Pharmaceuticals Ltd., which currently holds PCT/SG2018/050277 and derivatives thereof including issued US Patent No. 11,141,402.

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The authors confirm that all methods were performed in accordance with all relevant guidelines and regulations. All the animal experiments were approved by the Institutional Animal Care and Use Committee of the Laboratory Animal Centre of Peking University Shenzhen Graduate School (permit YW); and ethical approval was obtained from Tsinghua Shenzhen International Graduate School (Number: 37, Year 2019). Informed consent for the use of the LUAD tissue samples and clinical data was obtained by Outdo Bio-tech Co., Ltd. (Shanghai, China).

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Zhang, S., Tan, Y.Q., Zhang, X. et al. TFF3 drives Hippo dependent EGFR-TKI resistance in lung adenocarcinoma. Oncogene (2024). https://doi.org/10.1038/s41388-024-03244-5

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Received:12 April 2024

Revised:26 November 2024

Accepted:29 November 2024

Published:10 December 2024

DOI:https://doi.org/10.1038/s41388-024-03244-5

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