Site-dependent Treg cell transcriptional reprograming in a metastatic colorectal cancer model holds prognostic significance

Aristin Revilla, S.; Verheem, A.; Frederiks, C. L.; Kim, Y.; Chalkiadakis, T.; Viergever, B.; Gyorffy, B.; Mocholi, E.; Kranenburg, O.; Prekovic, S.; Coffer, P. J.

doi:10.1101/2025.03.13.643028

bioRxiv · 10.1101/2025.03.13.643028

Site-dependent Treg cell transcriptional reprograming in a metastatic colorectal cancer model holds prognostic significance

Aristin Revilla, S.·Verheem, A.·Frederiks, C. L.·Kim, Y.·Chalkiadakis, T.·Viergever, B.·Gyorffy, B.·Mocholi, E.·Kranenburg, O.·Prekovic, S.·Coffer, P. J.

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Abstract

In colorectal cancer (CRC), tumor-infiltrating regulatory T (Treg) cells suppress anti-tumor immunity, promoting immune evasion and tumor progression. Effective therapies require selectively targeting tumor-infiltrating Treg (TI-Treg) cells while preserving systemic Treg cells, necessitating insight into their adaptations within the tumor microenvironment. Here, CRC-organoids were implanted in the liver of Foxp3eGFP mice to investigate location-specific phenotypic differences in TI-Treg cells. Tumor tissue exhibited an increased proportion of Treg cells and a decrease of effector CD4 and CD8 T cells compared to matched healthy tissue. RNA sequencing of Treg cells isolated from the spleen, primary liver tumor transplant, or metastases identified gene expression profiles previously associated with CRC-related Treg cells in patients. Location-specific differences included elevated expression of WNT-pathway genes in peritoneal TI-Treg cells compared to liver counterparts. Higher expression of genes upregulated in liver TI-Treg cells correlated with poor CRC prognosis. Splenic Treg cells from tumor-bearing mice displayed distinct transcriptional profiles from both their healthy counterparts and TI-Treg cells, suggesting they represent a distinct CD4+ population. Taken together, these findings highlight TI-Treg cells heterogeneity across different tumor sites and the distinct nature of splenic Treg cells in tumor-bearing hosts.

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Aristin Revilla, S., Verheem, A., Frederiks, C. L., Kim, Y., Chalkiadakis, T., Viergever, B., Gyorffy, B., Mocholi, E., Kranenburg, O., Prekovic, S., Coffer, P. J.. 2025-03-15. Site-dependent Treg cell transcriptional reprograming in a metastatic colorectal cancer model holds prognostic significance. https://doi.org/10.1101/2025.03.13.643028

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