Supporting data for ""Genipin-Mediated Suppression of Inflammatory Macrophage Influx Reduces Postoperative Recurrence in Hepatocellular Carcinoma"
Surgical resection is the primary treatment regimen for early- to intermediate-stage hepatocellular carcinoma (HCC). However, postoperative recurrence affects up to 70% of patients within five years after surgery, significantly impairing postoperative patient prognosis. To date, no systemic therapeutics have been approved to prevent HCC recurrence, highlighting the urgent need to develop effective adjuvant postoperative interventions. The postoperative tumor microenvironment (TME), enriched with growth factors and inflammatory cytokines from liver regeneration and inflammation, facilitates residual tumor cell growth. Macrophages within the TME play a critical role in promoting tumor recurrence through angiogenesis and immunosuppression, with increased infiltration associated with higher recurrence risks. In this study, we investigated genipin, a compound derived from Gardenia jasminoides used in traditional Chinese medicine, as a novel therapeutic agent to suppress HCC recurrence by targeting postoperative macrophage influx.
An orthotopic mouse model of HCC recurrence was developed to replicate human postoperative HCC conditions, involving orthotopic tumor implantation followed by surgical resection. Genipin, administered orally at 25 mg/kg and 50 mg/kg, dose-dependently delayed postoperative recurrence and reduced tumor burden, Genipin exhibited minimal cytotoxicity against HCC cells in vitro, suggesting its effects are mediated through TME modulation rather than direct tumor cell killing. Specifically, genipin significantly reduced the infiltration of pro-inflammatory CD11b+F4/80+Ly6C+ macrophages into the postoperative liver by interfering with CCL2/CCR2 mediated macrophage migration. While postoperative adoptive transfer of macrophages accelerated tumor recurrence and abolished the therapeutic effects of genipin.
Mechanistically, genipin was identified as a natural PPARγ agonist. By activating PPARγ, genipin suppressed NF-κB p65-driven transcriptional programs that govern both CCR2 expression on macrophages and CCL2 secretion by hepatocytes. Through dual downregulation on the chemokine ligand production and receptor expression, genipin achieved robust inhibition of CCL2/CCR2 chemotaxis-mediated macrophage infiltration into the postoperative liver microenvironment and thereby inhibited HCC recurrence.
Finally, multiplex immunohistochemistry analysis on HCC human samples revealed that macrophage infiltration in the adjacent liver combined with vascular integrity of primary HCC could effectively stratify patients with higher recurrence risks. We highlight the adjacent liver tissue derived biomarkers with superior predictive efficacy for postoperative recurrence, underscoring their capacity in reflecting the postoperative microenvironment that harbors residual HCC cells.
