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Methotrexate-loaded tumour-cell-derived microvesicles can relieve biliary obstruction in patients with extrahepatic cholangiocarcinoma


Most patients with cholangiocarcinoma (CCA) develop extrahepatic malignant biliary obstructions, which require palliative drainage to normalize bilirubin levels and to improve the patients’ overall survival. Here, we report that the infusion of methotrexate-containing plasma-membrane microvesicles derived from apoptotic human tumour cells into the bile-duct lumen of patients with extrahepatic CCA mobilized and activated neutrophils and relieved biliary obstruction in 25% of the patients. Neutrophil recruitment by the microvesicles was associated with an increase in uridine diphosphate glucose and complement C5, and led to the degradation of the stromal barrier of CCA. The microvesicles induced pyroptosis of CCA cells through a gasdermin E-dependent pathway, and their intracellular contents released upon CCA-cell death activated patient-derived macrophages into producing proinflammatory cytokines, which attracted a secondary wave of neutrophils to the tumour site. Our findings suggest a possible treatment for the alleviation of obstructive extrahepatic CCA with few adverse effects, and highlight the potential of tumour-cell-derived microvesicles as drug carriers for antitumour therapies.

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Fig. 1: Effective treatment of obstructive CCA with MTX–TMPs.
Fig. 2: MTX–TMP perfusion attracts antitumor neutrophils.
Fig. 3: Direct attraction of neutrophils by UDPG in MTX–TMPs.
Fig. 4: MTX–TMPs induce CCA-cell pyroptosis.
Fig. 5: MTX–TMP perfusion triggers a secondary wave of recruiting neutrophils.
Fig. 6: MTX–TMP perfusion-recruited neutrophils display an antitumour phenotype.

Data availability

The main data supporting the results in this study are available within the paper and its Supplementary Information. The raw and analysed datasets generated during the study are too large to be publicly shared, but are available for research purposes from the corresponding authors on reasonable request.


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This work was supported by National Natural Science Foundation of China (81788101, 81661128007, 81530080, 81773062 and 91942314), the Chinese Academy of Medical Sciences Initiative for Innovative Medicine (CAMS-I2M) (2017-I2M-1-001 and 2016-I2M-1-007).

Author information




B.H., Y.L. and X.W. conceived the project. Y.L., Yunfeng Gao, Hui Zhang, N.Z., P.X., J.W., Yuan Gao, X.J., X.L., J.L., Y.Z., K.T., J.M., Huafeng Zhang and J.X. performed the experiments. B.H., Y.L., F.Y., W.T. and X.W. developed methodology. B.H., Y.L., Yunfeng Gao, Hui Zhang and X.W. performed data analysis. B.H. and Y.L. wrote the manuscript.

Corresponding authors

Correspondence to Yuying Liu, Ximo Wang or Bo Huang.

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Competing interests

B.H. is the inventor on patent no. ZL201110241369.8, owned by Hubei Soundny (Sheng-Qi-An) Biotech, which covers the preparation and use of drug-packaging tumour-cell-derived microparticles in cancer therapies. The other authors declare no competing interests.

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Supplementary information

Supplementary Information

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Reporting Summary

Supplementary Methods

Clinical research protocol.

Supplementary Methods

Appendix to clinical research protocol.

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Gao, Y., Zhang, H., Zhou, N. et al. Methotrexate-loaded tumour-cell-derived microvesicles can relieve biliary obstruction in patients with extrahepatic cholangiocarcinoma. Nat Biomed Eng 4, 743–753 (2020).

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