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Activation of the Notch1-c-myc-VCAM1 signalling axis initiates liver progenitor cell-driven hepatocarcinogenesis and pulmonary metastasis

Oncogene volume 41, pages 2340–2356 (2022)Cite this article

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Abstract

The cellular origin of hepatocellular carcinomas (HCC) and the role of Notch1 signalling in HCC initiation are controversial. Herein, we establish Notch1 as a regulator of HCC development and progression. Clinically, high Notch1 expression correlates with enhanced cancer progression, elevated lung metastasis, increased cancer stem cell (CSC)-like cells’ gene signature expression, and poor overall survival in HCC patients. Notch1 intracellular domain (N1ICD) overexpression spontaneously transforms rat liver progenitor cells (LPC) into CSC-like cells (WBN1ICD C5) under a selective growth environment, while orthotopic injection of these cells generates liver tumors and spontaneous pulmonary metastasis in an isogenic rat model. Mechanistically, the elevated Notch1 activity increases c-myc expression, which then transcriptionally upregulates VCAM1 expression to activate macrophage dependent HCC transendothelial migration. In vivo, silencing c-myc prohibits the tumorigenicity of WBN1ICD C5 cells, while depletion of VCAM1 reduces spontaneous lung metastasis without affecting primary WBN1ICD C5 orthotopic liver tumor growth. Importantly, depletion of macrophage or blockade of macrophage VCAM1 binding receptor α4β1-integrin reduces the number of WBN1ICD C5 lung nodules in an experimental metastasis model. Overall, our work discovers that the Notch1-c-myc-VCAM1 signaling axis initiates LPC-driven hepatocarcinogenesis and metastasis, providing a preclinical model for HCC study and therapeutic targets for an improved HCC treatment.

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Fig. 1: High Notch1 expression stratifies HCC patients into those with enhanced tumor size, increased cancer progression, and poor overall survival.
Fig. 2: N1ICD overexpression spontaneously transforms rat oval progenitor cells into CSC-like cells under selective growth process.
Fig. 3: Depletion of c-myc inhibits the tumorigenicity of WBN1ICD C1/C5 cells both in vitro and in vivo.
Fig. 4: Activation of the Notch1 signaling stimulates the spontaneous pulmonary metastasis of WBN1ICD C5 cells in an orthotopic rat model.
Fig. 5: Depletion of VCAM1 in WBN1ICD C5 cells inhibits the pulmonary metastasis without affecting primary tumor formation and growth.
Fig. 6: Notch1 overexpression enhances c-myc expression to up-regulate VCAM1 expression transcriptionally.
Fig. 7: Depletion of VCAM1 or macrophage inhibits the metastatic ability of WBN1ICD C5 cells in vivo.
Fig. 8: Schematic representation of the role of Notch1-mediated signalling pathway in promoting c-myc driven cancer stemness and facilitating pulmonary metastasis via VCAM1 dependent macrophage activation.

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Data availability

All the data supporting the findings of this study are available within the article and its supplementary information files and from the corresponding author upon reasonable request.

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Acknowledgements

Some of the illustrations were created using Biorender.com. This work was supported by grants from the National Natural Science Foundation of China (81972255, 82173195, 81920108028, 81872142, 81772597, 82173229, 81702904, 81972262), Guangdong Natural Science Foundation (2021A1515010095, 712055485047), Guangzhou Science and Technology Program Key Project (201904020008), Sun Yat-sen University Clinical Research 5010 Program (2018008), Science and Technology Program of Guangzhou (202102010326), Sun Yat-sen University Fundamental Research Funds (19ykpy113), the Key Training Program for Young Scholars of Sun Yat-sen University (18ykzd07), grant from the Key Laboratory of Malignant Tumor Molecular Mechanism and Translational Medicine of Guangzhou Bureau of Science and Information Technology ([2013]163), the Key Laboratory of Malignant Tumor Gene Regulation and Target Therapy of Guangdong Higher Education Institutes (KLB09001), Guangdong Science and Technology Department (2020A0505100029, 2020B1212060018, 2020B1212030004), the Guangdong Translational Medicine Public Platform of Guangdong Province (4202037).

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  1. These authors contributed equally: Wen-Rui Wu, Xiang-De Shi, Fa-Peng Zhang.

Authors and Affiliations

  1. Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, 510120, China

    Wen-Rui Wu, Xiang-De Shi, Fa-Peng Zhang, Ke Zhu, Rui Zhang, Xian-Huan Yu, Yu-Fei Qin, Shun-Peng He, Hou-Wei Fu, Lei Zhang, Hong Zeng, Man-Sheng Zhu, Lei-Bo Xu & Chao Liu

  2. Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Guangdong-Hong Kong Joint Laboratory for RNA medicine, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, 510120, China

    Wen-Rui Wu, Xiang-De Shi, Fa-Peng Zhang, Ke Zhu, Rui Zhang, Xian-Huan Yu, Yu-Fei Qin, Shun-Peng He, Hou-Wei Fu, Lei Zhang, Hong Zeng, Man-Sheng Zhu, Lei-Bo Xu, Ping-Pui Wong & Chao Liu

  3. Medical Research Center, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, 510120, China

    Ping-Pui Wong

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Contributions

WRW, XDS, and FPZ carried out the majority of the experiments and contributed equally to the paper; KZ, YFQ, SPH, HWF, LZ, and MSZ assisted the animal experiments; FPZ and HZ carried out the RNA-seq database and immunohistochemistry analysis; RZ and XHY provided technical advice. LBX, PPW, and CL conceived the study, co-supervised the research, and co-wrote the manuscript.

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Correspondence to Lei-Bo Xu, Ping-Pui Wong or Chao Liu.

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All animal procedures were approved by the animal care and use committee of Sun Yat-sen University.

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Wu, WR., Shi, XD., Zhang, FP. et al. Activation of the Notch1-c-myc-VCAM1 signalling axis initiates liver progenitor cell-driven hepatocarcinogenesis and pulmonary metastasis. Oncogene 41, 2340–2356 (2022). https://doi.org/10.1038/s41388-022-02246-5

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