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Cancer-cell stiffening via cholesterol depletion enhances adoptive T-cell immunotherapy

Nature Biomedical Engineering volume 5, pages 1411–1425 (2021)Cite this article

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Abstract

Malignant transformation and tumour progression are associated with cancer-cell softening. Yet how the biomechanics of cancer cells affects T-cell-mediated cytotoxicity and thus the outcomes of adoptive T-cell immunotherapies is unknown. Here we show that T-cell-mediated cancer-cell killing is hampered for cortically soft cancer cells, which have plasma membranes enriched in cholesterol, and that cancer-cell stiffening via cholesterol depletion augments T-cell cytotoxicity and enhances the efficacy of adoptive T-cell therapy against solid tumours in mice. We also show that the enhanced cytotoxicity against stiffened cancer cells is mediated by augmented T-cell forces arising from an increased accumulation of filamentous actin at the immunological synapse, and that cancer-cell stiffening has negligible influence on: T-cell-receptor signalling, production of cytolytic proteins such as granzyme B, secretion of interferon gamma and tumour necrosis factor alpha, and Fas-receptor–Fas-ligand interactions. Our findings reveal a mechanical immune checkpoint that could be targeted therapeutically to improve the effectiveness of cancer immunotherapies.

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Fig. 1: Cholesterol is enriched in the plasma membrane of cancer cells.
Fig. 2: Cancer-cell stiffness can be manipulated via supplementation or depletion of cholesterol in the cell membrane.
Fig. 3: Cancer-cell softness impairs T-cell-mediated cytotoxicity in vitro and in vivo.
Fig. 4: Cancer-cell stiffening by MeβCD enhances the efficacy of ACT immunotherapy.
Fig. 5: Cancer-cell stiffening has negligible influence on biochemical cancer-cell killing pathways mediated by T cells.
Fig. 6: Enhanced cytotoxicity against stiffened cancer cells is mediated by T-cell forces.
Fig. 7: Illustration of mechanical immuno-suppression induced by the softness of cancer cells.

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Data availability

The data supporting the results in this study are available within the paper and its Supplementary Information. Source data for the figures are provided with this paper.

Code availability

Source code for the custom ImageJ macro for F-actin analysis, cellular-force calculation and deformability cytometry analysis, and custom MATLAB scripts for optical-tweezer data collection and data post-processing are available from the corresponding author on reasonable request. Shape-Out (version 2.7.4) for deformability cytometry analysis is available at https://github.com/ZELLMECHANIK-DRESDEN/ShapeOut2.

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Acknowledgements

We thank R. Guiet (EPFL) for assistance on image analysis; W. Li (West China Hospital) for histological analyses of human biopsies; the EPFL Bioimaging and Optics Platform, Center of PhenoGenomics, Flow Cytometry Core Facility, Protein Production and Structure Core Facility, and Histology Core Facility for technical support; S. M. Leitão and B. Ghadiani (EPFL) for technical assistance on AFM measurement; P. Müller (Max Planck Institute) for assistance on analysing deformability cytometry data; D. J. Irvine (MIT) for providing IL-15SA construct, B16F10-OVA, and 4T1-Fluc-tdTomato cell lines; P. Romero (UNIL) for providing MC38-HER2 and Me275-HER2 cell lines; D. Trono (EPFL) for providing HEK293T cells and pLKO.1 vector; E. Amstad (EPFL) for providing access to a rheometer; F. Stellacci, M. de Palma, A. Cont, and A. Persat (EPFL) for discussion. This work was supported in part by the European Research under the ERC grant agreements MechanoIMM (805337) and ROBOCHIP (714609), the Swiss National Science Foundation (Project grant 315230_173243), the Swiss Cancer Research Foundation (No. KFS-4600-08-2018), Kristian Gerhard Jebsen Foundation, Anna Fuller Fund Grant, and École Polytechnique Fédérale de Lausanne (EPFL). A.K. acknowledges funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement no. 754354. M.G. was supported by the Chinese Scholarship Council (CSC) (No. 201808320453).

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Authors and Affiliations

  1. Institute of Materials Science and Engineering, École polytechnique fédérale de Lausanne (EPFL), Lausanne, Switzerland

    Kewen Lei, Armand Kurum & Li Tang

  2. Institute of Mechanical Engineering, EPFL, Lausanne, Switzerland

    Murat Kaynak & Mahmut Selman Sakar

  3. Institute of Bioengineering, EPFL, Lausanne, Switzerland

    Lucia Bonati, Veronika Cencen, Min Gao, Yu-Qing Xie, Yugang Guo, Mélanie T. M. Hannebelle, Georg E. Fantner, Mahmut Selman Sakar & Li Tang

  4. Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

    Yulong Han & Ming Guo

  5. Department of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu, China

    Yangping Wu & Guanyu Zhou

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  1. Kewen Lei
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Contributions

K.L. and L.T. conceived the study and designed the experiments. K.L., A.K., M.K., L.B., Y.H., V.C., M.G., Y.-Q.X., Y.G., M.T.M.H., Y.W., G.Z., M.G., G.E.F., and M.S.S. performed the experiments. K.L., A.K., L.T., M.S.S., M.G., and G.E.F. analysed the data. L.T. supervised the project. K.L., A.K., and L.T. wrote the manuscript. All authors edited the manuscript.

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Correspondence to Li Tang.

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Lei, K., Kurum, A., Kaynak, M. et al. Cancer-cell stiffening via cholesterol depletion enhances adoptive T-cell immunotherapy. Nat Biomed Eng 5, 1411–1425 (2021). https://doi.org/10.1038/s41551-021-00826-6

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