Disruption of SLFN11 Deficiency-Induced CCL2 Signaling and Macrophage M2 Polarization Potentiates Anti-PD-1 Therapy Efficacy in Hepatocellular Carcinoma

BACKGROUND & AIMS: The therapeutic effect of immune checkpoint inhibitors (ICIs) is poor in hepatocellular carcinoma (HCC) and varies greatly among individuals. Schlafen (SLFN) family members have important functions in immunity and oncology, but their roles in cancer immunobiology remain un-clear. We aimed to investigate the role of the SLFN family in immune responses against HCC. METHODS: Transcriptome analysis was performed in human HCC tissues with or without response to ICIs. A humanized orthotopic HCC mouse model and a co-culture system were constructed, and cytometry by time-of -flight technology was used to explore the function and mechanism of SLFN11 in the immune context of HCC. RESULTS: SLFN11 was significantly up-regulated in tumors that responded to ICIs. Tumor-specific SLFN11 deficiency increased the infiltration of immunosuppressive macrophages and aggravated HCC progression. HCC cells with SLFN11 knockdown promoted macrophage migration and M2-like po-larization in a C-C motif chemokine ligand 2-dependent manner, which in turn elevated their own PD-L1 expression by activating the nuclear factor-KB pathway. Mechanistically, SLFN11 suppressed the Notch pathway and C-C motif chemo-kine ligand 2 transcription by binding competitively with tripartite motif containing 21 to the RNA recognition motif 2 domain of RBM10, thereby inhibiting tripartite motif containing 21-mediated RBM10 degradation to stabilize RBM10 and promote NUMB exon 9 skipping. Pharmacologic antagonism of C-C motif chemokine receptor 2 potentiated the antitumor ef-fect of anti-PD-1 in humanized mice bearing SLFN11 knock-down tumors. ICIs were more effective in patients with HCC with high serum SLFN11 levels. CONCLUSIONS: SLFN11 serves as a critical regulator of microenvironmental immune proper-ties and an effective predictive biomarker of ICIs response in HCC. Blockade of C-C motif chemokine ligand 2/C-C motif chemokine receptor 2 signaling sensitized SLFN11low HCC pa-tients to ICI treatment.

Automated summary

This study found that SLFN11, a member of the Schlafen (SLFN) family, plays a critical role in the response to immune checkpoint inhibitors (ICIs) in hepatocellular carcinoma (HCC) patients. SLFN11 deficiency promotes the infiltration of immunosuppressive macrophages and aggravates HCC progression. SLFN11 regulates the immune microenvironment by suppressing the Notch pathway and C-C motif chemokine ligand 2 transcription, and is associated with the response to ICIs. Inhibition of the C-C motif chemokine ligand 2/C-C motif chemokine receptor 2 signaling pathway enhances the therapeutic efficacy of ICIs in HCC patients with low SLFN11 levels.