FGF19/SOCE/NFATc2 signaling circuit facilitates the self-renewal of liver cancer stem cells

<b>Background & Aims:</b> Liver cancer stem cells (LCSCs) mediate therapeutic resistance and correlate with poor outcomes in patients with hepatocellular carcinoma (HCC). Fibroblast growth factor (FGF)-19 is a crucial oncogenic driver gene in HCC and correlates with poor prognosis. However, whether FGF19 signaling regulates the self-renewal of LCSCs is unknown. <b>Methods:</b> LCSCs were enriched by serum-free suspension. Self-renewal of LCSCs were characterized by sphere formation assay, clonogenicity assay, sorafenib resistance assay and tumorigenic potential assays. Ca²⁺ image was employed to determine the intracellular concentration of Ca²⁺. Gain- and loss-of function studies were applied to explore the role of FGF19 signaling in the self-renewal of LCSCs. <b>Results:</b> FGF19 was up-regulated in LCSCs, and positively correlated with certain self-renewal related genes in HCC. Silencing FGF19 suppressed self-renewal of LCSCs, whereas overexpressing FGF19 facilitated CSCs-like properties <i>via</i> activation of FGF receptor (FGFR)-4 in none-LCSCs. Mechanistically, FGF19/FGFR4 signaling stimulated store-operated Ca²⁺ entry (SOCE) through both the PLCγ and ERK1/2 pathways. Subsequently, SOCE-calcineurin signaling promoted the activation and translocation of nuclear factors of activated T cells (NFAT)-c2, which transcriptionally activated the expression of stemness-related genes (<i>e.g., NANOG</i>, <i>OCT4</i> and <i>SOX2</i>), as well as <i>FGF19</i>. Furthermore, blockade of FGF19/FGFR4-NFATc2 signaling observably suppressed the self-renewal of LCSCs. <b>Conclusions:</b> FGF19/FGFR4 axis promotes the self-renewal of LCSCs <i>via</i> activating SOCE/NFATc2 pathway; in turn, NFATc2 transcriptionally activates FGF19 expression. Targeting this signaling circuit represents a potential strategy for improving the therapeutic efficacy of HCC.