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Cysteine cathepsins L and X differentially modulate interactions between myeloid-derived suppressor cells and tumor cells

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Abstract

Increased proteolytic activity of cysteine cathepsins has long been known to facilitate malignant progression, and it has also been associated with tumor-promoting roles of myeloid-derived suppressor cells (MDSCs). Consequently, cysteine cathepsins have gained much attention as potential targets for cancer therapies. However, cross-talk between tumor cells and MDSCs needs to be taken into account when studying the efficacy of cathepsin inhibitors as anti-cancer agents. Here, we demonstrate the potential of the MDA-MB-231 breast cancer cell line to generate functional MDSCs from CD14+ cells of healthy human donors. During this transition to MDSCs, the overall levels of cysteine cathepsins increased, with the largest responses for cathepsins L and X. We used small-molecule inhibitors of cathepsins L and X (i.e., CLIK-148, Z9, respectively) to investigate their functional impact on tumor cells and immune cells in this co-culture system. Interactions with peripheral blood mononuclear cells reduced MDA-MB-231 cell invasion, while inhibition of cathepsin X activity by Z9 restored invasion. Inhibition of cathepsin L activity using CLIK-148 resulted in significantly increased CD8+ cytotoxicity. Of note, inhibition of cathepsins L and X in separate immune or tumor cells did not promote these functional changes. Together, our findings underlie the importance of tumor cell–immune cell interactions in the evaluation of the anti-cancer potential of cysteine cathepsin inhibitors.

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Abbreviations

Cat:

Cathepsin(s)

CFSE:

Carboxyfluorescein succinimidyl ester

GM-CSF:

Granulocyte-macrophage colony-stimulating factor

IL:

Interleukin

MDSCs:

Myeloid-derived suppressor cells

PBMCs:

Peripheral blood mononuclear cells

PBS:

Phosphate-buffered saline

PG:

Prostaglandin

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Acknowledgements

The authors would like to acknowledge Matthew Bogyo (Stanford University) for the DCG-04 probe, and Christopher Berrie for critical review of the manuscript before submission.

Funding

This study was supported by the Research Agency of Republic of Slovenia (Grants P4-0127, J4-1776, J4-8227 and J4-6811, to JK).

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Authors and Affiliations

  1. Department of Pharmaceutical Biology, Faculty of Pharmacy, University of Ljubljana, Aškerčeva cesta 7, 1000, Ljubljana, Slovenia

    Tanja Jakoš, Anja Pišlar, Urša Pečar Fonović & Janko Kos

  2. Department of Biotechnology, Jožef Stefan Institute, Ljubljana, Slovenia

    Janko Kos

  3. Department for Therapeutic Services, Blood Transfusion Centre of Slovenia, Ljubljana, Slovenia

    Urban Švajger

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  1. Tanja Jakoš
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Contributions

TJ performed the experiments and cooperated with UŠ on the work with the PBMCs. TJ analyzed the data and prepared the figures. TJ, AP, UPF, and JK designed the framework for the study. AP and UPF offered technical support. TJ drafted the paper, and the other co-authors (AP, UPF, UŠ, JK) revised and complemented its content.

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Correspondence to Janko Kos.

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Jakoš, T., Pišlar, A., Pečar Fonović, U. et al. Cysteine cathepsins L and X differentially modulate interactions between myeloid-derived suppressor cells and tumor cells. Cancer Immunol Immunother 69, 1869–1880 (2020). https://doi.org/10.1007/s00262-020-02592-x

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