Collagen density regulates the activity of tumor-infiltrating T cells

Dorota E. Kuczek, Anne Mette Hvid Larsen, Marie-Louise Thorseth, Marco Carretta, Adrija Kalvisa, Majken Siersbæk, Ana Micaela C. Simões, Anne Roslind, Lars Henning Engelholm, Elfriede Noessner, Marco Donia, Inge Marie Svane, Per thor Straten, Lars Grøntved, Daniel Hargbøl Madsen

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Abstract

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
ISSN2051-1426
DOI
StatusUdgivet - 2019

Emneord

  • Tumor microenvironment
  • T cell activity
  • Extracellular matrix
  • Immune modulation
  • 3D culture

Citer dette

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