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Cellular and Molecular Biology

Macrophage-derived CCL23 upregulates expression of T-cell exhaustion markers in ovarian cancer

Abstract

Background

Macrophages are an important component of the tumour immune microenvironment (TME) and can promote tumour growth and metastasis. Macrophage-secreted chemokine-ligand-23 (CCL23) induces ovarian cancer cell migration via chemokine-receptor 1 (CCR1). However, the effect of CCL23 on other immune cells in the TME is unknown.

Methods

CCL23 levels were measured by ELISA. The expression of surface markers in exhaustion assays was quantified by flow cytometry. Signalling pathways were identified by phosphokinase array and validated by western blot.

Results

Ascites from patients with high-grade serous ovarian cancer (HGSC) contain high levels of CCL23. Similarly, significantly higher CCL23 levels were found in plasma from HGSC patients compared to healthy individuals. RNA-seq analysis of ovarian cancer tissues from TCGA showed that expression of CCL23 correlated with the presence of macrophages. In tissues with high levels of CCL23 and macrophage content, the fraction of CD8 + T cells expressing exhaustion markers CTLA-4 and PD-1 were significantly higher compared to low-level CCL23 tissues. In vitro, CCL23 induced upregulation of immune checkpoint proteins on CD8 + T cells, including CTLA-4, TIGIT, TIM-3 and LAG-3 via phosphorylation of GSK3β in CD8 + T cells.

Conclusions

Our data suggest that CCL23 produced by macrophages contributes to the immune-suppressive TME in ovarian cancer by inducing an exhausted T-cell phenotype.

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Fig. 1: CCL23 is elevated in plasma and ascites from patients with ovarian cancer.
Fig. 2: Higher CCL23 expression correlates with the increase in exhausted CD8 + T cells and macrophage infiltration in TCGA.
Fig. 3: CCL23 induces expression of exhaustion markers in CD8 + T cells.
Fig. 4: CCL23-induced T-cell signalling.

Data availability

RNA-seq data for patients with ovarian cancer were retrieved from TCGA, which is publicly available at https://portal.gdc.cancer.gov/.

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Funding

This work was supported by the Mary Lake Polan Gynecologic Oncology Endowment for Research (OD), the Vivian Scott Fellowship in Gynecologic Oncology (OD).

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Contributions

KK, VK and OD designed experiments; KK carried out and analyzed experiments; KK, VK and OD prepared the figures and wrote the initial draft of the manuscript; all authors edited the manuscript; VK and OD provided supervision.

Corresponding author

Correspondence to Oliver Dorigo.

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Competing interests

KK and VK have no competing interests. OD has served on Advisory Boards for Merck, Eisai, PACT, GSK, IMV, Genentech. OD received funding for clinical research from AstraZeneca, IMV, Millenium, Pharmamar, Genentech, Bioeclipse.

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Patient samples were collected under an approved Institutional Review Board (IRB) protocol at Stanford University and the University of Pennsylvania. Informed consent for healthy plasma was obtained by Innovative Research (Novi, MI). This study was performed in accordance with the Declaration of Helsinki.

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Kamat, K., Krishnan, V. & Dorigo, O. Macrophage-derived CCL23 upregulates expression of T-cell exhaustion markers in ovarian cancer. Br J Cancer (2022). https://doi.org/10.1038/s41416-022-01887-3

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