24-Norursodeoxycholic acid reshapes immunometabolism in CD8+ T cells and alleviates hepatic inflammation

Abstract

Background & aims: 24-Norursodeoxycholic acid (NorUDCA) is a novel therapeutic bile acid used to treat immune-mediated cholestatic liver diseases, such as primary sclerosing cholangitis (PSC), where dysregulated T cells including CD8⁺ T cells contribute to hepatobiliary immunopathology. We hypothesized that NorUDCA may directly modulate CD8⁺ T cell function thus contributing to its therapeutic efficacy.

Methods: NorUDCA's immunomodulatory effects were first studied in Mdr2^-/- mice, as a cholestatic model of PSC. To differentiate NorUDCA's immunomodulatory effects on CD8⁺ T cell function from its anticholestatic actions, we also used a non-cholestatic model of hepatic injury induced by an excessive CD8⁺ T cell immune response upon acute non-cytolytic lymphocytic choriomeningitis virus (LCMV) infection. Studies included molecular and biochemical approaches, flow cytometry and metabolic assays in murine CD8⁺ T cells in vitro. Mass spectrometry was used to identify potential CD8⁺ T cell targets modulated by NorUDCA. The signaling effects of NorUDCA observed in murine cells were validated in circulating T cells from patients with PSC.

Results: NorUDCA demonstrated immunomodulatory effects by reducing hepatic innate and adaptive immune cells, including CD8⁺ T cells in the Mdr2^-/- model. In the non-cholestatic model of CD8⁺ T cell-driven immunopathology induced by acute LCMV infection, NorUDCA ameliorated hepatic injury and systemic inflammation. Mechanistically, NorUDCA demonstrated strong immunomodulatory efficacy in CD8⁺ T cells affecting lymphoblastogenesis, expansion, glycolysis and mTORC1 signaling. Mass spectrometry identified that NorUDCA regulates CD8⁺ T cells by targeting mTORC1. NorUDCA's impact on mTORC1 signaling was further confirmed in circulating PSC CD8⁺ T cells.

Conclusions: NorUDCA has a direct modulatory impact on CD8⁺ T cells and attenuates excessive CD8⁺ T cell-driven hepatic immunopathology. These findings are relevant for treatment of immune-mediated liver diseases such as PSC.

Lay summary: Elucidating the mechanisms by which 24-norursodeoxycholic acid (NorUDCA) works for the treatment of immune-mediated liver diseases, such as primary sclerosing cholangitis, is of considerable clinical interest. Herein, we uncovered an unrecognized property of NorUDCA in the immunometabolic regulation of CD8⁺ T cells, which has therapeutic relevance for immune-mediated liver diseases, including PSC.

Keywords: (phospho-) proteome; CD8(+) T cells; Immune-mediated liver disease; Primary Sclerosing Cholangitis; bile acid; mass spectrometry; metabolism.