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SMAD4 endows TGF-β1-induced highly invasive tumor cells with ferroptosis vulnerability in pancreatic cancer

Acta Pharmacologica Sinica volume 45, pages 844–856 (2024)Cite this article

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is an extremely aggressive malignancy prone to recurrence and metastasis. Studies show that tumor cells with increased invasive and metastatic potential are more likely to undergo ferroptosis. SMAD4 is a critical molecule in the transforming growth factor β (TGF-β) pathway, which affects the TGF-β-induced epithelial-mesenchymal transition (EMT) status. SMAD4 loss is observed in more than half of patients with PDAC. In this study, we investigated whether SMAD4-positive PDAC cells were prone to ferroptosis because of their high invasiveness. We showed that SMAD4 status almost determined the orientation of transforming growth factor β1 (TGF-β1)-induced EMT via the SMAD4-dependent canonical pathway in PDAC, which altered ferroptosis vulnerability. We identified glutathione peroxidase 4 (GPX4), which inhibited ferroptosis, as a SMAD4 down-regulated gene by RNA sequencing. We found that SMAD4 bound to the promoter of GPX4 and decreased GPX4 transcription in PDAC. Furthermore, TGF-β1-induced high invasiveness enhanced sensitivity of SMAD4-positive organoids and pancreas xenograft models to the ferroptosis inducer RAS-selective lethal 3 (RSL3). Moreover, SMAD4 enhanced the cytotoxic effect of gemcitabine combined with RSL3 in highly invasive PDAC cells. This study provides new ideas for the treatment of PDAC, especially SMAD4-positive PDAC.

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Fig. 1: SMAD4 determines highly invasive status induced by TGF-β1 in PDAC.
Fig. 2: SMAD4 affects ferroptosis vulnerability in highly invasive pancreatic cancer.
Fig. 3: SMAD4 inhibits GPX4 expression in pancreatic cancer.
Fig. 4: SMAD4 affects ferroptosis vulnerability by targeting GPX4 in highly invasive pancreatic cancer.
Fig. 5: SMAD4-determined the sensitivity to ferroptosis in highly invasive cancer cells is further confirmed in PDOs and pancreas xenograft model.
Fig. 6: SMAD4 potentiates the cytotoxic effect of gemcitabine combined with RSL3 in highly invasive pancreatic cancer.
Fig. 7: GPX4 is associated with a poor prognosis in SMAD4-positive PDAC.

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Acknowledgements

The work was supported by the National Natural Science Foundation of China (No. 82173281, 82141129, 82173282, 82172577 and 82172948); the Shanghai Municipal Science and Technology Commission (20ZR1471100); the Clinical and Scientific Innovation Project of Shanghai Hospital Development Center (SHDC12018109); the Scientific Innovation Project of Shanghai Education Committee (2019-01-07-00-07-E00057); the Shanghai Municipal Commission of Health and Family Planning (No. 2018YQ06); the Excellence Project of Shanghai Municipal Health Commission (20224Z0006); the Sailing Project of Science and Technology Commission of Shanghai Municipality (22YF1409000).

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  1. These authors contributed equally: Hai-di Chen, Zeng Ye, Hai-feng Hu

Authors and Affiliations

  1. Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai, 200032, China

    Hai-di Chen, Zeng Ye, Hai-feng Hu, Gui-xiong Fan, Yu-heng Hu, Zheng Li, Bo-rui Li, Shun-rong Ji, Chen-jie Zhou, Xiao-wu Xu, Xian-jun Yu & Yi Qin

  2. Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, 200032, China

    Hai-di Chen, Zeng Ye, Hai-feng Hu, Gui-xiong Fan, Yu-heng Hu, Zheng Li, Bo-rui Li, Shun-rong Ji, Chen-jie Zhou, Xiao-wu Xu, Xian-jun Yu & Yi Qin

  3. Shanghai Pancreatic Cancer Institute, Shanghai, 200032, China

    Hai-di Chen, Zeng Ye, Hai-feng Hu, Gui-xiong Fan, Yu-heng Hu, Zheng Li, Bo-rui Li, Shun-rong Ji, Chen-jie Zhou, Xiao-wu Xu, Xian-jun Yu & Yi Qin

  4. Pancreatic Cancer Institute, Fudan University, Shanghai, 200032, China

    Hai-di Chen, Zeng Ye, Hai-feng Hu, Gui-xiong Fan, Yu-heng Hu, Zheng Li, Bo-rui Li, Shun-rong Ji, Chen-jie Zhou, Xiao-wu Xu, Xian-jun Yu & Yi Qin

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Contributions

Conceptualization, YQ, HDC, ZY and HFH; Methodology, HDC, YHH, GXF and ZL; Statistical analysis, HDC, BRL, SRJ and CJZ; Investigation, HDC, YQ, XWX and XJY; Writing, HDC and ZY; Supervision, XJY, XWX and YQ. All authors read and approved the final paper.

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Correspondence to Xiao-wu Xu, Xian-jun Yu or Yi Qin.

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This study was approved by the Institutional Research Ethics Committee of Fudan University Shanghai Cancer Centre (ethical code: 050432-4-1212B) and written informed consent was obtained from all patients.

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Chen, Hd., Ye, Z., Hu, Hf. et al. SMAD4 endows TGF-β1-induced highly invasive tumor cells with ferroptosis vulnerability in pancreatic cancer. Acta Pharmacol Sin 45, 844–856 (2024). https://doi.org/10.1038/s41401-023-01199-z

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