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P21-activated kinase 2-mediated β-catenin signaling promotes cancer stemness and osimertinib resistance in EGFR-mutant non-small-cell lung cancer

Oncogene volume 41, pages 4318–4329 (2022)Cite this article

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Abstract

Osimertinib (AZD9291) is a third-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), used for treating patients with advanced non-small-cell lung cancer (NSCLC) harboring EGFR-activating mutations or the resistant T790M mutation. However, acquired resistance to osimertinib is inevitable in EGFR-mutant NSCLC. By employing a global mass spectrometry-based phosphoproteomics approach, we identified that the activated p21-activated kinase 2 (PAK2)/β-catenin axis acts as a driver of osimertinib resistance. We found that PAK2 directly phosphorylates β-catenin and increases the nuclear localization of β-catenin, leading to the increased expression and transcriptional activity of β-catenin, which in turn enhances cancer stem-like properties and osimertinib resistance. Moreover, we revealed that HER3 as an upstream regulator of PAK2, drives the activation of PAK2/β-catenin pathways in osimertinib-resistant cells. The clinical relevance of these findings was further confirmed by examining tissue specimens from patients with EGFR-mutant NSCLC. The results demonstrated that the levels of HER3, phospho-PAK2 (p-PAK2) and β-catenin in the tissues from patients with EGFR-mutant NSCLC, that had relapsed after treatment with osimertinib, were elevated compared to those of the corresponding untreated tissues. Additionally, the high levels of HER3, p-PAK2 and β-catenin correlated with shorter progression-free survival (PFS) in patients with EGFR-TKI-treated NSCLC. We additionally observed that the suppression of PAK2 via knockdown or pharmacological targeting with PAK inhibitors markedly restored the response of osimertinib-resistant NSCLC cells to osimertinib both in vitro and in vivo. In conclusion, these results indicated that the PAK2-mediated activation of β-catenin is important for osimertinib resistance and targeting the HER3/PAK2/β-catenin pathway has potential therapeutic value in NSCLCs with acquired resistance to osimertinib.

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Fig. 1: Quantitative phosphoproteomic analysis of osimertinib resistance in NSCLC cells.
Fig. 2: Activation of β-catenin is associated with osimertinib resistance and cancer stemness.
Fig. 3: PAK2 regulates the phosphorylation and transcriptional activity of β-catenin in osimertinib-resistant NSCLC cells.
Fig. 4: PAK2 induces osimertinib resistance by promoting β-catenin-dependent stemness in NSCLCs.
Fig. 5: Increased HER3 contributes to osimertinib resistance and regulates PAK2 phosphorylation.
Fig. 6: HER3/PAK2/β-catenin signaling is activated in clinical samples of osimertinib-resistant NSCLC and is associated with poor clinical outcome in patients with EGFR-mutant NSCLC.
Fig. 7: Targeting PAK2 synergizes with osimertinib in suppressing the growth of osimertinib-resistant NSCLC.

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Funding

This work was supported by the National Natural Science Foundation of China (81972194, 82172810, 81772825); Guangdong Natural Science Funds (2019A1515010113); The Science and Technology Program of Guangzhou (202201010839); Medical Scientific Research Foundation of Guangdong (B2020165); Discipline construction project of Guangdong Medical University (4SG22005G).

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  1. These authors contributed equally: Yanmei Yi, Pan Li, Yuanfeng Huang.

Authors and Affiliations

  1. School of Basic Medical Sciences, Guangdong Medical University, Zhanjiang, Guangdong, China

    Yanmei Yi, Siwen Fan, Xinwu Lv & Zhiwei He

  2. Affiliated Cancer Hospital & Institute of Guangzhou Medical University, Guangzhou Key Laboratory of “Translational Medicine on Malignant Tumor Treatment”, Guangzhou, Guangdong, China

    Pan Li, Yuanfeng Huang, Danyang Chen, Jun Wang, Minqiang Yang, Shanshan Zeng, Jin Deng, Kai Luo & Hao Liu

  3. China-America Cancer Research Institute, Guangdong Medical University, Dongguan, Guangdong, China

    Zhiwei He

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Contributions

HL, ZH, KL, and JD designed the experiments, analyzed the data, and drafted the manuscript and figures. YY, PL, YH, DC, SF, JW, MY, SZ, and XL detected the cells’ biological function, performed IHC analyses and animal experiments. All authors read and approved the final manuscript.

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Correspondence to Kai Luo, Zhiwei He or Hao Liu.

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Yi, Y., Li, P., Huang, Y. et al. P21-activated kinase 2-mediated β-catenin signaling promotes cancer stemness and osimertinib resistance in EGFR-mutant non-small-cell lung cancer. Oncogene 41, 4318–4329 (2022). https://doi.org/10.1038/s41388-022-02438-z

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