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Plasmalemma vesicle-associated protein promotes angiogenesis in cholangiocarcinoma via the DKK1/CKAP4/PI3K signaling pathway

Oncogene volume 40, pages 4324–4337 (2021)Cite this article

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Abstract

Cholangiocarcinoma (CCA) is aggressive and has poor clinical outcomes because of typically delayed diagnosis and a lack of effective non-surgical therapeutic options. Recent studies have shown that plasmalemma vesicle-associated protein (PLVAP) is related to angiogenesis in various tumors, and in vivo PLVAP targeting therapy has been proven effective against hepatocellular carcinoma and pancreatic cancer. The goal of this study was to determine the potential therapeutic utility of targeting PLVAP and thus angiogenesis in CCA and explore the underlying molecular mechanisms. We found that the PLVAP expression levels were significantly higher in CCA tissues when compared with matched adjacent non-tumor tissues obtained from a total of 90 CCA patients; higher expression levels of PLVAP were associated with shorter overall survival of patients. In addition, overexpression of PLVAP was associated with higher micro-vessel density in CCA tissues. In a PLVAP overexpressing CCA patient-derived xenograft model, a novel humanized anti-PLVAP antibody in combination with Gemcitabine plus Cisplatin was significantly inhibited tumor growth. Molecular analysis of CCA cells co-cultured with human umbilical vascular endothelial cells or human hepatic sinusoidal endothelial cells showed that Dickkopf-related protein 1 (DKK1) secreted by CCA cells activated the PI3K/Akt pathway after binding to its receptor, cytoskeleton-associated protein 4 (CKAP4), resulting in the upregulation of PLVAP. Thus, CCA cells increased the angiogenic potency of endothelial cells in a paracrine fashion. Consistently, patients bearing CKAP4 and PLVAP overexpressing tumors had a poor prognosis. In conclusion, the DKK1/CKAP4/PI3K/PLVAP pathway increases angiogenesis in CCA and is therefore a potential anti-angiogenic target.

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Fig. 1: PLVAP is an angiogenic marker and indicates poor prognosis in CCA patients.
Fig. 2: PLVAP is an anti-angiogenic target for CCA.
Fig. 3: CCA cells increase the angiogenic potential of endothelial cells (ECs) in a paracrine manner.
Fig. 4: CCA-secreted DKK1 is a key regulator of PLVAP expression and angiogenic potential of ECs.
Fig. 5: DKK1 binds to the CKAP4 receptor and up-regulates PLVAP expression by activating the PI3K/Akt pathway.
Fig. 6: High CKAP4 and PLVAP expression levels are associated with poor overall survival of CCA patients underwent surgical resection.

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Acknowledgements

We thank Jungang Zhao, Yifan Tong, Ziyan Chen, and Mingxun Wang for discussing the paper. We also appreciate Sina Zhang for her help with statistical analysis and bioinformatics analysis. We are deeply thankful to Prof Lewis R.Roberts (Mayo Clinic, USA) for valuable advice and discussions on the paper. This work was supported by the National Natural Science Foundation of China (Grant No. 81201953, 81772628, 82072685 to GC and 81703310 to YW); the Joint Projects of National Health Council and Zhejiang Province (No.WKJ-ZJ-1706 to GC); the Natural Science Foundation of Zhejiang Province (No.LY17H160047 to GC); the Public Projects of Zhejiang Province (No.2016C37127 to ZPY and 2018C37114 to YW).

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  1. These authors contributed equally: Yi Wang, Haitao Yu, Xiaozai Xie

Authors and Affiliations

  1. Division of Preventive Medicine, School of Public Health and Management, Wenzhou Medical University, Wenzhou, China

    Yi Wang

  2. Key Laboratory of Diagnosis and Treatment of Severe Hepato-Pancreatic Diseases of Zhejiang Province, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China

    Haitao Yu, Xiaozai Xie, Tuo Deng, Longyun Ye, Lijun Wu, Qiandong Zhu, Junjian Li, Yihu Zheng, Zhengping Yu & Gang Chen

  3. Department of Hepatobiliary Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China

    Haitao Yu, Xiaozai Xie, Tuo Deng, Longyun Ye, Lijun Wu, Qiandong Zhu, Junjian Li, Yihu Zheng, Zhengping Yu & Gang Chen

  4. Department of Gastroenterology, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing, Jiangsu, China

    Xiwei Ding

  5. Department of Infectious Diseases, Shandong Provincial Hospital affiliated to Shandong University, Jinan, China

    Zhen Yang

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Contributions

Conception and design: GC, ZPY. Development of methodology: GC, YW, YHZ. Acquisition of data (provided animals, acquired and managed patients, provided facilities, etc.): GC, XZX, HTY, LYY, LJW, QDZ, JJL, XWD, ZY. Analysis and interpretation of data (e.g., statistical analysis, biostatistics, computational analysis): GC, YW, XZX, HTY, LYY, LJW, XWD, ZY, TD. Writing, review, and revision of the paper: GC, YW, ZPY, TD. Administrative, technical, or material support (i.e., reporting or organizing data, constructing databases): GC, XZX, HTY, LJW, QDZ, JJL, XWD, ZY, YHZ. Study supervision: GC, YW.

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Correspondence to Yi Wang or Gang Chen.

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Wang, Y., Yu, H., Xie, X. et al. Plasmalemma vesicle-associated protein promotes angiogenesis in cholangiocarcinoma via the DKK1/CKAP4/PI3K signaling pathway. Oncogene 40, 4324–4337 (2021). https://doi.org/10.1038/s41388-021-01844-z

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