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Cellular and Molecular Biology

Stromal localization of inactive CD8+ T cells in metastatic mismatch repair deficient colorectal cancer

British Journal of Cancer volume 130, pages 213–223 (2024)Cite this article

Subjects

Abstract

Background

The determinants of metastasis in mismatch repair deficiency with high levels of microsatellite instability (MSI-H) in colorectal cancer (CRC) are poorly understood. Here, we hypothesized that distinct immune and stromal microenvironments in primary tumors may discriminate between non-metastatic MSI-H CRC and metastatic MSI-H CRC.

Methods

We profiled 46,727 single cells using high-plex imaging mass cytometry and analyzed both differential cell type abundance, and spatial distribution of fibroblasts and immune cells in primary CRC tumors with or without metastatic capacity. We validated our findings in a second independent cohort using immunohistochemistry.

Results

High-plex imaging mass cytometry and hierarchical clustering based on microenvironmental markers separated primary MSI-H CRC tumors with and without metastatic capacity. Primary tumors with metastatic capacity displayed a high stromal content and low influx of CD8+ T cells, which expressed significantly lower levels of markers reflecting proliferation (Ki67) and antigen-experience (CD45RO) compared to CD8+ T cells in non-metastatic tumors. CD8+ T cells showed intra-epithelial localization in non-metastatic tumors, but stromal localization in metastatic tumors, which was validated in a second cohort.

Conclusion

We conclude that localization of phenotypically distinct CD8+ T cells within stroma may predict metastasis formation in MSI-H CRC.

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Fig. 1: TME markers distinguish non-metastatic from metastatic colorectal tumors.
Fig. 2: Cell-type annotation reveals differential microenvironment composition between metastatic and non-metastatic primary CRC tumors.
Fig. 3: Metastatic MSI-H tumors lack proliferating antigen-experienced CD8+ T cells.
Fig. 4: Spatial analysis reveals distinct localization of cell-types in the tumor microenvironment.
Fig. 5: Cytotoxic T cells reside in high stromal content in metastatic MSI-H tumors.

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Data availability

All data are publicly available in Zenodo repository (https://doi.org/10.5281/zenodo.8341225).

Code availability

All code are available in Github (https://github.com/VercoulenLab).

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Acknowledgements

The authors thank all patients involved in the current study.

Funding

This work was supported by the Dutch Cancer Society (KWF/Alpe d’HuZes #UU-10660), and the Dutch Scientific Organization (cancergenomicscenter.nl, NWO Gravitation 024.001.028). Funders had no role in study design, data acquisition, analysis, and interpretation.

Author information

Author notes

  1. These authors contributed equally: Emre Küçükköse, Matthijs J. D. Baars, Onno Kranenburg, Yvonne Vercoulen.

Authors and Affiliations

  1. Division of Imaging and Cancer, Laboratory Translational Oncology, University Medical Center Utrecht, Utrecht, The Netherlands

    Emre Küçükköse, Inne H. M. Borel Rinkes, Jeanine M. L. Roodhart, Jamila Laoukili & Onno Kranenburg

  2. Center for Molecular Medicine, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands

    Matthijs J. D. Baars, Mojtaba Amini, Evelien Floor & Yvonne Vercoulen

  3. UCyTOF.nl, Center for Molecular Medicine, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands

    Mojtaba Amini & Yvonne Vercoulen

  4. Division of Imaging and Cancer, Department of Medical Oncology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands

    Suzanna J. Schraa, Guus M. Bol, Jeanine M. L. Roodhart & Miriam Koopman

  5. Utrecht Platform for Organoid Technology, Utrecht University, Utrecht, The Netherlands

    Onno Kranenburg

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Contributions

Study design: EK, MJDB, OK, YV. Experimental work: EK, MJDB, MA, EF. Resources: SS, GB, JMLR, MK. Data interpretation: EK, MJDB, IHMBR, JL, OK, YV. Study supervision: OK, YV. Writing manuscript: EK, MJDB, OK, YV.

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Correspondence to Onno Kranenburg or Yvonne Vercoulen.

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Küçükköse, E., Baars, M.J.D., Amini, M. et al. Stromal localization of inactive CD8+ T cells in metastatic mismatch repair deficient colorectal cancer. Br J Cancer 130, 213–223 (2024). https://doi.org/10.1038/s41416-023-02500-x

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