Intraoral Gavage of Second-Generation Probiotic Lactobacillus Reuteri Releasing IFN-β (LR-IFN-β) Mitigates Intestinal Irradiation Toxicity and Improves Survival During Whole Abdomen Irradiation (WAI)

Abstract

We sought to establish a method by which to overcome the toxicity of WAI to facilitate clinical application in Ovarian Cancer patients. We irradiated C57BL/6J mice to 19.75 Gy WAI and assessed the primary endpoint of overall survival (OS). In a separate experiment, mice were irradiated to 12 Gy WAI and intestinal barrier integrity was compared between groups: control (0 Gy), irradiation only, 12 Gy + LR, 12 Gy + IFN-β, and 12 Gy + LR-IFN-β. Luminex assay of plasma and intestinal cells were also assayed at day 5 after WAI for radiation-induced inflammatory cytokines, and fecal matter was analyzed for LR-IFN-β clearance and levels of the LR-derived IFN-β gene from day 1 to 5 in control non-irradiated mice. Moreover, fluorescent beads were intraorally administered three hours prior to sacrifice at days 2 or 5 after WAI, and blood was assayed for beads. Mice receiving LR-IFN-β (109 bacteria in 100 mL of saline) 24-hours following a single fraction of 19.75 Gy WAI showed improved OS compared to control irradiated mice (p = 0.03). LR-IFN-β gavage maintained intestinal barrier integrity (p < 0.05) by stimulating intestinal stem cells regenerations (improved levels of Lgr5+ cells, occludin, and I-CAM; p < 0.05), and reduced levels of intestinal pro-inflammatory cytokines, including IFN-γ (p = 0.0261), IL-3 (p < 0.0020) and IL-17 (p < 0.0070). There was no significant effect of control LR or intraperitoneal injection of IFN-β protein at 24 hours after WAI. Detectable levels of LR-IFN-β bacteria were also cleared from fecal matter by day three via colony assay and rt-PCR, with no detectable growth of LR-IFN-β in blood from gavaged irradiated mice (13.5 Gy WAI). LR-IFN-β is both a feasible and effective radiation mitigator that could potentially improve the management of ovarian cancer patients. Furthermore, the subsequent addition of platinum/taxane-based chemotherapy to the combination of WAI and LR-IFN-β should reduce tumor volume while protecting the intestine and thus improve overall survival in ovarian cancer patients.