UDPG-P2Y14R axis licenses cDC2 differentiation

Conventional type-2 dendritic cells (cDC2) are essential for orchestrating adaptive immunity in inflammatory skin diseases, yet the extracellular cues that guide their differentiation remain incompletely understood. We recently identified the metabolite UDP-glucose (UDPG) as an endogenous ligand for the purinergic receptor P2Y14R and demonstrated that this axis is required for normal cDC2 development both in vitro and in vivo. To uncover the molecular programs controlled by UDPG-P2Y14R signaling, we performed RNA-seq on immature cDC2 differentiated in vitro under defined conditions. Bone-marrow cells were harvested from 8-12-week-old wild-type (C57BL/6J) or global P2Y14R-knockout mice (P2ry14-/-). cDC2 precursors were then induced with 50 uM 2-mercaptoethanol, 20 ng/mL recombinant mouse GM-CSF, and 100 ng/mL FLT3-L and cultured for 6 days in RPMI-1640 supplemented with 10 % FBS. Cultures were continuously exposed to 100 uM UDPG or vehicle (PBS). The following experimental groups were generated: (1) WT + UDPG, (2) P2ry14-/- + UDPG (n = 3 biological replicates per group).