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

Abstract

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24-Nor-ursodeoxycholic acid counteracts TH17/Treg imbalance and ameliorates intestinal inflammation by restricting glutaminolysis in differentiating TH17 cells

“…However, by focusing on TH17 cells we further identified that through restricting glutaminolysis NorUDCA inhibits mTORC1 activity which is directly controlled by α-Ketoglutarate, a key metabolic intermediate biosynthesized along the glutaminolysis cascade (figure 7). Revisiting the mass spectrometry data set of NorUDCA-treated CD8 + T cells published in our previous study [11], we found that in accordance with our observation in TH17 cells the abundance of glutaminolysis rate-limiting enzyme GLS2 (Glutaminase 2) was also reduced. This suggests that GLS2 might be a potential metabolic checkpoint for NorUDCA's modulatory actions of glutamine metabolism in adaptive immune cells, thus provides an interesting direction for future investigations.…”

“…Cell surface and intracellular FACS stains for ex vivo isolated immune cells were performed at 4°C for 30min as previously described [11] using antibodies listed in Supplementary table 1. Intracellular staining of cytokines and transcription factors of in vitro cultured T H 17 cells was performed as described in [11].…”

“…Cell surface and intracellular FACS stains for ex vivo isolated immune cells were performed at 4°C for 30min as previously described [11] using antibodies listed in Supplementary table 1. Intracellular staining of cytokines and transcription factors of in vitro cultured T H 17 cells was performed as described in [11]. Intracellular staining of pRPS6 Ser235/236 of in vitro cultured T H 17 cells was performed by fixing for 10 min at 37°C with Cytofix (BD), permeabilized 20min on ice with methanol and stained with the indicated phospho-Abs for 30min in the dark at 4°C in PBS/2% FCS.…”

“…Concentrations of NorUDCA (500μM) for in vitro experiments were selected to mimic the in vivo serum bile acid level of mice on 0.5% (wt/wt) NorUDCA supplemented diet [11] (also as indicated in figure 2B). Concentrations of NorUDCA selected for in vitro experiments were previously tested not affecting viability in T cells [11]. Rapamycin (mTOR inhibitor; Cell signaling) was used as 100nM.…”

“…NorUDCA is currently under evaluation for its long-term effect on PSC in a phase III trial (NCT03872921). Recently we have uncovered a yet unrecognized immunoregulatory role of NorUDCA in directly modulating CD8 + T cell metabolism and immune response through blunting mTORC1 activity during hepatic immunopathology, profoundly distinct from its parent compound UDCA [11]. This offers exciting entry points of exploring NorUDCA's additional therapeutic potentials in applications of T cell mediated diseases, in addition to liver disorders.…”