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Targeting TLK2 inhibits the progression of gastric cancer by reprogramming amino acid metabolism through the mTOR/ASNS axis

Cancer Gene Therapy volume 30, pages 1485–1497 (2023)Cite this article

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Abstract

Several recent studies have suggested that TLKs are related to tumor progression. However, the function and mechanism of action of TLK2 in gastric cancer (GC) remain elusive. In this study, TLK2 was found to be significantly upregulated in patients with GC and was identified as an independent prognostic factor for GC. Consistently, TLK2 knockdown markedly reduced the aggressiveness of GC, whereas its overexpression had the opposite effect. IP-MS revealed that the effects of TLK2 on GC were mainly associated with metabolism reprogramming. TLK2 knockdown suppressed amino acid synthesis by downregulating the mTORC1 pathway and ASNS expression in GC cells. Mechanistically, mTORC1 directly interacts with the ASNS protein and inhibits its degradation. Further experiments validated that the ASNS protein was degraded via ubiquitination instead of autophagy. Inhibiting and activating the mTORC1 pathway can upregulate and downregulate ASNS ubiquitination, respectively, and the mTORC1 pathway can reverse the regulatory effects of TLK2 on ASNS. Furthermore, TLK2 was found to regulate the mRNA expression of ASNS. TLK2 directly interacted with ATF4, a transcription factor of ASNS, and promoted its expression. The kinase inhibitor fostamatinib significantly inhibited the proliferative, invasive, and migratory capabilities of GC cells by inhibiting TLK2 activity. Altogether, this study reveals a novel functional relationship between TLK2 and the mTORC1/ASNS axis in GC. Therefore, TLK2 may serve as a potential therapeutic target for GC.

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Fig. 1: TLK2 expression was upregulated in GC, and high TLK2 expression was associated with a worse prognosis.
Fig. 2: TLK2 knockdown inhibited the colony-forming, migratory and invasive capabilities of GC cells in vitro.
Fig. 3: TLK2 regulated the proliferation and apoptosis of GC cells in vitro and in vivo through a metabolic pathway.
Fig. 4: mTORC1 and ASNS were intermediate molecules of TLK2 regulating amino acid metabolism.
Fig. 5: mTORC1 interacted with ASNS and inhibited its ubiquitin-mediated degradation.
Fig. 6: ATF4 and mTORC1 participate in the dual regulating effects of TLK2-mediated regulation of ASNS.
Fig. 7: Fostamatinib inhibited the proliferative, invasive and migratory capabilities of GC cells by targeting TLK2.
Fig. 8: TLK2 promoted the binding between mTORC1 and ASNS, thereby inhibiting the degradation and enhancing the stability of the ASNS protein.

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Data availability

All data relevant to the study are included in the article or uploaded as online Supplementary materials. Source data will be made available upon direct request to the corresponding author.

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Acknowledgements

We thank Bullet Edits Limited for the linguistic editing and proofreading of the manuscript.

Funding

This research was funded by the Key Programs of the Educational Commission of Anhui Province (the scientific research project chaired by Mingliang Wang from May 2023 to April 2025 and the project number has not yet been given) and the National Natural Science Foundation (grant no. 81874063).

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  1. These authors contributed equally: Mingliang Wang, Jing Li.

Authors and Affiliations

  1. General Surgery Department, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, China

    Mingliang Wang, Jing Li, Xiaodong Yang, Qiang Yan, Huizhen Wang, Xin Xu, Yida Lu, Deguan Li, Yigao Wang, Ruochuan Sun, Shangxin Zhang, Yonghong Zhang, Zhen Zhang, Futao Meng & Yongxiang Li

  2. Department of Surgical Oncology, The First Affiliated Hospital of Bengbu Medical College, No. 287 Changhuai Road, Bengbu, China

    Futao Meng

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Contributions

MW and YL designed this study. MW and FM wrote the manuscript. MW, JL, QY, HW, XX and YL performed the experiments. JL, YZ, SZ and RS assisted with cell culture and in vivo experiments. XY, DL, YW and ZZ were responsible for clinical sample collection. All authors have read and approved the final version of this manuscript.

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Correspondence to Futao Meng or Yongxiang Li.

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All procedures involving human participants were reviewed and approved by the Ethics Committee of Anhui Medical University (ethical code: 20180323). All patients/participants provided written informed consent to participate in this study.

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Wang, M., Li, J., Yang, X. et al. Targeting TLK2 inhibits the progression of gastric cancer by reprogramming amino acid metabolism through the mTOR/ASNS axis. Cancer Gene Ther 30, 1485–1497 (2023). https://doi.org/10.1038/s41417-023-00653-8

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