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CRMP2 is a therapeutic target that suppresses the aggressiveness of breast cancer cells by stabilizing RECK

Oncogene volume 39, pages 6024–6040 (2020)Cite this article

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Abstract

Metastatic breast cancer is characterized by high mortality and limited therapeutic target. During tumor metastasis, cytoskeletal reorganization is one of the key steps in the migration and invasion of breast cancer cells. Collapsin response mediator protein 2 (CRMP2) is a cytosolic phosphoprotein that plays an important role in regulating cytoskeletal dynamics. Previous researches have reported that altered CRMP2 expression is associated with breast cancer progression, but the underlying mechanism remains poorly understood. Here, we show that CRMP2 expression is reduced in various subtypes of breast cancers and negatively correlated with lymphatic metastasis. Overexpression of CRMP2 significantly inhibits invasion and stemness in breast cancer cells, while downregulation of CRMP2 promotes cell invasion, which is not required for tubulin polymerization. Mechanistic studies demonstrate that CRMP2 interacts with RECK, prevents RECK degradation, which, in turn, blocks NF-κB and Wnt signaling pathways. Furthermore, we find that phosphorylation of CRMP2 at T514 and S522 remarkably abolishes its functions to bind with RECK and to inhibit cell invasion. Pharmacologic rescue of CRMP2 expression suppressed breast cancer metastasis in vitro and in vivo and stimulated a synergetic effect with FN-1501 that induces CRMP2 dephosphorylation. Collectively, this study highlights the potential of CRMP2 as a therapeutic target in breast cancer metastasis and reveals a distinct mechanism of CRMP2.

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Fig. 1: CRMP2 is significantly downregulated in breast cancers and is negatively correlated with metastatic ability.
Fig. 2: CRMP2 suppresses breast cancer invasion in a microtubule-independent manner.
Fig. 3: CRMP2 inhibits breast cancer metastasis without disturbing tumor growth in vivo.
Fig. 4: CRMP2 blocks both NF-κB and Wnt signaling pathways in breast cancer cells.
Fig. 5: CRMP2 interacts with RECK to prevent its degradation.
Fig. 6: CRMP2 phosphorylation at T514 and S522 sites impedes its ability to inhibit invasion in breast cancer cells.
Fig. 7: CRMP2 is a binding target of NA which shows an irreversible anti-invasion activity.
Fig. 8: Elevated protein expression and decreased phosphorylation level of CRMP2 generate synergistic effects in suppressing tumor metastasis.

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Acknowledgements

The authors are grateful to Professor Jinfeng Liu for his kind help with in silico drug screening for this research. We appreciate Professor Ahmed Ashour Ahmed for his kind gifts of CRMP2 plasmids. We thank Ms. Liuyi Zhong for her kind help with in vivo imaging experiments.

Funding

This work was supported by the National Natural Science Foundation of China (Nos. 81872899 and 81903625), the National Science & Technology Major Project (Nos. 2017ZX09101003-005-023, 2017ZX09301014, and 2018ZX09711001-003-007), Social Development Project of Jiangsu Provincial Science and Technology Department (BE2018711), Natural Science Foundation of Jiangsu Province (BK20190563), and the “Double First-Class” University project (CPU2018GY17).

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  1. These authors contributed equally: Binyan Lin, Yongxu Li

Authors and Affiliations

  1. State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, Department of Physiology, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing, 210009, People’s Republic of China

    Binyan Lin, Yongxu Li, Tiepeng Wang, Yangmin Qiu, Zhenzhong Chen, Kai Zhao & Na Lu

  2. School of Pharmacy, Nanjing University of Chinese Medicine, Xianlin Avenue No. 138, Nanjing, 210023, People’s Republic of China

    Binyan Lin

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Contributions

BL performed the transwell assay, western blots, immunofluorescence analysis, Co-IP assay, IHC assay, colony formation experiments, DARTS, CETSA assay, primer design, and RT-PCR assay. YL performed the pull-down assay, animal assay, sphere-formation assay, and IF assay. TW, YQ, and ZC contributed to the CAM assay and plasmid extraction. KZ and NL analyzed the data and wrote the paper. All authors read and approved the final paper.

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Correspondence to Kai Zhao or Na Lu.

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The present study was approved by the Experimental Animal Ethics Committee of China Pharmaceutical University.

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Lin, B., Li, Y., Wang, T. et al. CRMP2 is a therapeutic target that suppresses the aggressiveness of breast cancer cells by stabilizing RECK. Oncogene 39, 6024–6040 (2020). https://doi.org/10.1038/s41388-020-01412-x

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