Background:
Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal malignancies, with pervasive resistance to Gemcitabine (Gem) and Nab-Paclitaxel (Nab) driving poor clinical outcomes. To identify novel therapeutic adjuncts, multiple cannabinoids(e.g. CBG, CBD, CBC) were screened across MIAPaCa-2, PANC-1, and patient-derived xenograft (PDX) CCT-4IT cell lines. Among them, Cannabigerol (CBG), a non-psychotropic cannabinoid, emerged as a potent candidate, yet its mechanistic role in chemo-sensitization remains underexplored.

Methods:
Cytotoxic synergy between CBG and chemotherapeutics (Gem, Nab Paclitaxel[Nab], and the triple combination CBG+Gem+Nab) was assessed in MIAPaCa-2, PANC-1 and PDX CCT-4IT cells in 2D and 3D cultures. Western blot analyses quantified alterations in key resistance and survival pathways (AXL, PI3K/AKT, MAPK/ERK, NF-κB, Notch1, PD-L1, and serotonergic receptors 5-HT1B/1D), along with markers of apoptosis, ferroptosis, and autophagy. Migration was evaluated via scratch assays. Translational efficacy was validated in PDAC xenograft mouse models treated with CBG alone or in combination with Gem or Nab.

Results:
Initial cannabinoid screening using 2D and 3D assays identified CBG as the most potent anticancer compound across MIAPaCa-2, PANC-1, and PDX CCT-4IT pancreatic cancer cell lines tested, and subsequent studies revealed that CBG exhibited strong synergy with both Gem and Nab, leading to broad suppression of key oncogenic signaling networks. The combination treatments markedly downregulated p-AXL, p-PI3K/p-AKT, p-MEK/p-ERK, NF-κB, Notch1, PD-L1, and the serotonergic receptors 5-HT1B/1D (p < 0.001), indicating effective disruption of major pathways driving chemoresistance. Mechanistic analyses further showed that CBG activated three complementary modes of programmed cell death: apoptosis, evidenced by increased Cytochrome-C release; ferroptosis, demonstrated by pronounced GPX4 suppression most notable in the CBG+Gem+Nab group; and autophagy, indicated by elevated LC3B expression. In addition to these cytotoxic mechanisms, CBG-based combinations significantly inhibited cell migration. Consistent with the in vitro findings, in vivo administration of CBG (100 mg/kg) together with Gem (12.5 mg/kg), or in the triple combination with Nab (5 mg/kg), resulted in superior tumor growth suppression and significantly improved survival compared with treatments using individual agents alone. No toxicity was observed in CBG and Gem combinations. Currently studies are being conducted with PDAC organoids to further support our results.

Learning Objectives:

• The audience will be able to understand the mechanism of action of CBG in pancreatic cancer
  
  
• The synergism of CBG with gemcitabine at a low chemotherapeutic dose and its mechanisms against aggressive pancreatic cancer will also be understood.