Cxcr3 promotes protection from colorectal cancer liver metastasis by driving NK cell infiltration and plasticity
Eleonora Russo, Chiara D’Aquino, Chiara Di Censo, Mattia Laffranchi, Luana Tomaipitinca, Valerio Licursi, Stefano Garofalo, Johann Promeuschel, Giovanna Peruzzi, Francesca Sozio, Anna Kaffke, Cecilia Garlanda, Ulf Panzer, Cristina Limatola, Christian A.J. Vosshenrich, Silvano Sozzani, Giuseppe Sciumè, Angela Santoni, Giovanni Bernardini
Eleonora Russo, Chiara D’Aquino, Chiara Di Censo, Mattia Laffranchi, Luana Tomaipitinca, Valerio Licursi, Stefano Garofalo, Johann Promeuschel, Giovanna Peruzzi, Francesca Sozio, Anna Kaffke, Cecilia Garlanda, Ulf Panzer, Cristina Limatola, Christian A.J. Vosshenrich, Silvano Sozzani, Giuseppe Sciumè, Angela Santoni, Giovanni Bernardini
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Research Article Immunology Oncology

Cxcr3 promotes protection from colorectal cancer liver metastasis by driving NK cell infiltration and plasticity

Abstract

The antimetastatic activity of NK cells is well established in several cancer types, but the mechanisms underlying NK cell metastasis infiltration and acquisition of antitumor characteristics remain unclear. Herein, we investigated the cellular and molecular factors required to facilitate the generation of an ILC1-like CD49a+ NK cell population within the liver metastasis (LM) environment of colorectal cancer (CRC). We show that CD49a+ NK cells had the highest cytotoxic capacity among metastasis-infiltrating NK cells in the MC38 mouse model. Furthermore, the chemokine receptor CXCR3 promoted CD49a+ NK cell accumulation and persistence in metastasis where NK cells colocalize with macrophages in CXCL9- and CXCL10-rich areas. By mining a published scRNA-seq dataset of a cohort of patients with CRC who were treatment naive, we confirmed the accumulation of CXCR3+NK cells in metastatic samples. Conditional deletion of Cxcr3 in NKp46+ cells and antibody-mediated depletion of metastasis-associated macrophages impaired CD49a+NK cell development, indicating that CXCR3 and macrophages contribute to efficient NK cell localization and polarization in LM. Conversely, CXCR3neg NK cells maintained a CD49a– phenotype in metastasis with reduced parenchymal infiltration and tumor killing capacity. Furthermore, CD49a+ NK cell accumulation was impaired in an independent SL4-induced CRC metastasis model, which fails to accumulate CXCL9+ macrophages. Together, our results highlight a role for CXCR3/ligand axis in promoting macrophage-dependent NK cell accumulation and functional sustenance in CRC LM.

Authors

Eleonora Russo, Chiara D’Aquino, Chiara Di Censo, Mattia Laffranchi, Luana Tomaipitinca, Valerio Licursi, Stefano Garofalo, Johann Promeuschel, Giovanna Peruzzi, Francesca Sozio, Anna Kaffke, Cecilia Garlanda, Ulf Panzer, Cristina Limatola, Christian A.J. Vosshenrich, Silvano Sozzani, Giuseppe Sciumè, Angela Santoni, Giovanni Bernardini

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Figure 1

The liver metastasis microenvironment reorganizes type 1 ILC compartment and deeply shapes their transcriptional profile.

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The liver metastasis microenvironment reorganizes type 1 ILC compartment...
(A) Left, representative contour plots of ILC1s (CD49a+CD49b–), CD49a–NK cells (CD49a–CD49b+) and CD49a+NK cells (CD49a+CD49b+) in healthy liver (HL), metastasis-free liver (MFL), and liver metastasis (LM) gated on lin(CD3CD19)–CD45+NK1.1+ NKp46+ cells. Right, histograms show mean ± SEM of ILC1, CD49a+ NK, and CD49a– NK cell frequency among CD45+ cells in HL, MFL, and LM of MC38-injected tumor-bearing mice. 5 independent experiments with at least 5 mice per group were performed. (n ≥ 15, ILC1 **P = 0.007; CD49a+NK ****P < 0.0001, 1-way ANOVA). (B) Top, gating scheme for cell populations isolated by fluorescence activated cell sorting. Pooled MFL and LM samples from at least 6 mice were collected in 3 independent sortings. Bottom, principal component analysis (PCA) from bulk RNA-seq. (C) Heatmaps showing DEGs involved in regulation of cellular functions. (D) Gene set variation analysis (GSVA) of MFL ILC1 and NK cells and LM CD49a– NK cells and CD49a+ NK showing modulation of pathways associated to cell proliferation (E2F targets, G2M checkpoint), activation (IL-15-treated NK cells, TGF-β1, and IFN-α signalling) and metabolism (glycolysis). (E) Representative FACS histogram plots of at least 3 independent analyses for cell surface receptor expression by LM CD49a+ NK cells, ILC1, and CD49a– NK cells.