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Targeting CLK4 inhibits the metastasis and progression of breast cancer by inactivating TGF-β pathway

Cancer Gene Therapy volume 29, pages 1168–1180 (2022)Cite this article

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A Correction to this article was published on 20 July 2022

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Abstract

Triple-negative breast cancer (TNBC) represents the most aggressive subtype of breast cancer that is highly resistant to current therapeutic options. According to the public databases Oncomine and KM plotter, the CLK4 expression is correlated with poor patient survival in TNBC, especially in mesenchymal-like TNBC (MES-TNBC) that has strong metastatic potential. Therefore, we investigated the potential involvement of CLK4 in the metastasis and progression of MES-TNBC. In the MES-TNBC cell lines, the CLK4 expression was elevated. Notably, the RNAi-mediated silencing of CLK4 reduced the expression of multiple epithelial-mesenchymal transition (EMT) genes that mediate metastasis. Furthermore, CLK4 silencing reduced both the invasive behaviors of the cultured cells and tumor metastasis in the mouse xenograft model. It is also noteworthy that CLK4 silencing repressed the invasive and cancer stem cell (CSC) properties that are induced by the TGF-β signaling. Importantly, the pharmacological inhibition of CLK4 potently repressed the invasion and proliferation of MES-TNBC cell lines and patient-derived cells, which demonstrates its clinical applicability. Collectively, our results suggest that CLK4 plays a crucial role in invasion and proliferation of MES-TNBC, especially in the processes that are induced by TGF-β. Also, this study characterizes CLK4 as a novel therapeutic target in breast cancer.

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Fig. 1: CLK4 expression is elevated in invasive breast cancers and is associated with poor overall survival in patients with MES-TNBC.
Fig. 2: CLK4 silencing impairs the proliferation and invasion of MES-TNBC cells.
Fig. 3: CLK4 silencing impairs TGF-β-mediated EMT in MES-TNBC cells.
Fig. 4: CLK4 silencing impairs TGF-β-mediated induction of CSC-like properties in MES-TNBC cells.
Fig. 5: CLK4 inhibition via ML167 impairs the proliferation, invasion, and chemoresistance of MDA-MB-231 cells.
Fig. 6: CLK4 inhibition via ML167 impairs the proliferation of primary cultured breast cancer cells.
Fig. 7: CLK4 silencing impairs the metastasis of MDA-MB-231 cells in vivo.

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Acknowledgements

This study was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. 2018R1A2A2A05022732) and Samsung Research Funding Center of Samsung Electronics (No. SRFC-MA1501-51).

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Authors and Affiliations

  1. Cancer Research Institute, Seoul National University, 101 Daehak-ro, Jongno-gu, Seoul, Republic of Korea

    Eunji Kang, Han-Byoel Lee & Wonshik Han

  2. Department of Oncology, MRC Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford, Oxford, UK

    Kanggeon Kim

  3. Department of Surgery, Seoul National University College of Medicine, 101 Daehak-ro, Jongno-gu, Seoul, Republic of Korea

    Sook Young Jeon, Ji Gwang Jung, Hong-Kyu Kim, Han-Byoel Lee & Wonshik Han

  4. Biomedical Research Institute, Seoul National University Hospital, 101 Daehak-ro, Jongno-gu, Seoul, Republic of Korea

    Han-Byoel Lee & Wonshik Han

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Contributions

EK and WH conceived the ideas. EK designed the studies, performed the experiments, and interpreted the data. EK and KG wrote and polished the manuscript. KG conducted literature search for setting up the experiments involving inhibitors. SYJ, JGJ, HKK, HBL and WH prepared the clinical samples and contributed to data analysis. All authors have read and approved the final manuscript.

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Correspondence to Wonshik Han.

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Kang, E., Kim, K., Jeon, S.Y. et al. Targeting CLK4 inhibits the metastasis and progression of breast cancer by inactivating TGF-β pathway. Cancer Gene Ther 29, 1168–1180 (2022). https://doi.org/10.1038/s41417-021-00419-0

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