Lactobacillus gallinarum-derived metabolites boost anti-PD1 efficacy in colorectal cancer by inhibiting regulatory T cells through modulating IDO1/Kyn/AHR axis

Abstract

ObjectiveGut microbiota is a key player in dictating immunotherapy response. We aimed to explore the immunomodulatory effect of probioticLactobacillus gallinarumand its role in improving anti-programmed cell death protein 1 (PD1) efficacy against colorectal cancer (CRC).DesignThe effects ofL. gallinarumin anti-PD1 response were assessed in syngeneic mouse models and azoxymethane/dextran sulfate sodium-induced CRC model. The change of immune landscape was identified by multicolour flow cytometry and validated by immunohistochemistry staining and in vitro functional assays. Liquid chromatography-mass spectrometry was performed to identify the functional metabolites.ResultsL. gallinarumsignificantly improved anti-PD1 efficacy in two syngeneic mouse models with different microsatellite instability (MSI) statuses (MSI-high for MC38, MSI-low for CT26). Such effect was confirmed in CRC tumourigenesis model.L. gallinarumsynergised with anti-PD1 therapy by reducing Foxp3+CD25+regulatory T cell (Treg) intratumoural infiltration, and enhancing effector function of CD8+T cells.L. gallinarum-derived indole-3-carboxylic acid (ICA) was identified as the functional metabolite. Mechanistically, ICA inhibited indoleamine 2,3-dioxygenase (IDO1) expression, therefore suppressing kynurenine (Kyn) production in tumours. ICA also competed with Kyn for binding site on aryl hydrocarbon receptor (AHR) and antagonised Kyn binding on CD4+T cells, thereby inhibiting Treg differentiation in vitro. ICA phenocopiedL. gallinarumeffect and significantly improved anti-PD1 efficacy in vivo, which could be reversed by Kyn supplementation.ConclusionL. gallinarum-derived ICA improved anti-PD1 efficacy in CRC through suppressing CD4+Treg differentiation and enhancing CD8+T cell function by modulating the IDO1/Kyn/AHR axis.L. gallinarumis a potential adjuvant to augment anti-PD1 efficacy against CRC.