Collagen density regulates the activity of tumor-infiltrating T cells

Dorota E Kuczek, Anne Mette H Larsen, Marie-Louise Thorseth, Marco Carretta, Adrija Kalvisa, Majken S Siersbæk, Ana Micaela C Simões, Anne Roslind, Lars H Engelholm, Elfriede Noessner, Marco Donia, Inge Marie Svane, Per Thor Straten, Lars Grøntved, Daniel H Madsen*

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BACKGROUND: Tumor progression is accompanied by dramatic remodeling of the surrounding extracellular matrix leading to the formation of a tumor-specific ECM, which is often more collagen-rich and of increased stiffness. The altered ECM of the tumor supports cancer growth and metastasis, but it is unknown if this effect involves modulation of T cell activity. To investigate if a high-density tumor-specific ECM could influence the ability of T cells to kill cancer cells, we here studied how T cells respond to 3D culture in different collagen densities.

METHODS: T cells cultured in 3D conditions surrounded by a high or low collagen density were imaged using confocal fluorescent microscopy. The effects of the different collagen densities on T cell proliferation, survival, and differentiation were examined using flow cytometry. Cancer cell proliferation in similar 3D conditions was also measured. Triple-negative breast cancer specimens were analyzed for the number of infiltrating CD8+ T cells and for the collagen density. Whole-transcriptome analyses were applied to investigate in detail the effects of collagen density on T cells. Computational analyses were used to identify transcription factors involved in the collagen density-induced gene regulation. Observed changes were confirmed by qRT-PCR analysis.

RESULTS: T cell proliferation was significantly reduced in a high-density matrix compared to a low-density matrix and prolonged culture in a high-density matrix led to a higher ratio of CD4+ to CD8+ T cells. The proliferation of cancer cells was unaffected by the surrounding collagen-density. Consistently, we observed a reduction in the number of infiltrating CD8+ T-cells in mammary tumors with high collagen-density indicating that collagen-density has a role in regulating T cell abundance in human breast cancer. Whole-transcriptome analysis of 3D-cultured T cells revealed that a high-density matrix induces downregulation of cytotoxic activity markers and upregulation of regulatory T cell markers. These transcriptional changes were predicted to involve autocrine TGF-β signaling and they were accompanied by an impaired ability of tumor-infiltrating T cells to kill autologous cancer cells.

CONCLUSIONS: Our study identifies a new immune modulatory mechanism, which could be essential for suppression of T cell activity in the tumor microenvironment.

OriginalsprogEngelsk
Artikelnummer68
TidsskriftJournal for immunotherapy of cancer
Vol/bind7
Udgave nummer1
Antal sider15
ISSN2051-1426
DOI
StatusUdgivet - 12. mar. 2019

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Kuczek, D. E., Larsen, A. M. H., Thorseth, M-L., Carretta, M., Kalvisa, A., Siersbæk, M. S., Simões, A. M. C., Roslind, A., Engelholm, L. H., Noessner, E., Donia, M., Svane, I. M., Straten, P. T., Grøntved, L., & Madsen, D. H. (2019). Collagen density regulates the activity of tumor-infiltrating T cells. Journal for immunotherapy of cancer, 7(1), [68]. https://doi.org/10.1186/s40425-019-0556-6