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 5

Macrophages showing TAM-like features accumulate in MC38-induced liver metastasis and produce cytokines and chemokines.

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Macrophages showing TAM-like features accumulate in MC38-induced liver m...
(A) Histograms show chemokine concentrations in LM homogenates from MC38- and SL4-injected mice by luminex assay performed in duplicate (n = 3; pg/mg of tissue lysate) (CCL3 *P = 0.01, CCL4 *P = 0.04, KC *P = 0.01, ***P = 0.0007, 2-tailed Student’s t test). (B) Top, quantification of CXCL9, CXCL10, and IL-15/IL-15Ra complex secretion in MFL and LM macrophage- and MC38 cell-conditioned supernatants (n ≥ 2). Bottom, mRNA levels of Il-15 and Il-15Rα expression by macrophages isolated from MFL and LM and by MC38 cells (n = 2). (C) mRNA levels of TGF-β1 expression by MC38- and SL4-derived LM macrophages and MC38 and SL4 cell lines (n = 4, *P = 0.02, 2-tailed Student’s t test). (D) Contour plots show macrophage subsets HL and MC38-derived MFL and LM. Histograms represent mean frequency ± SEM of F4/80int (CD11bhiF4/80int) and F4/80hi cells (CD11bhiF4/80hi) gated on live lin–CD45+Ly6Clow/–Gr1– from at least 12 total mice. (*P = 0.02, ****P < 0.0001, 1-way ANOVA). (E) Histogram plots show expression of Arg1, MMR, PD-L1, LAP-1, CX3CR1, and MHCII in F4/80int and F4/80hi from MC38-induced LM (Arg1 *P = 0.017, MMR **P = 0.001, PD-L1 *P = 0.02, LAP-1 *P = 0.014, CX3CR1 ***P = 0.0005, MHCII ***P = 0.0001, 2-tailed Student’s t test) (F) Heatmaps showing DEGs from bulk RNA-seq of sorted F4/80hi (violet) and F4/80int (green) macrophages from MC38-derived LM.