Su1246 SPECIFIC BACTERIA ISOLATED FROM HUMAN BREAST MILK BOLSTER INTESTINAL EPITHELIAL BARRIER FUNCTION, AND AMELIORATE DYSFUNCTION INDUCED BY SALMONELLA ENTERICA INFECTION

Background: Sulforaphane (SFN) is a plant derived glucosinolate activated from a precursor metabolite to an active isothiocyonate by myrosinase activity of gut microbiota.This phytochemical has numerous purported health benefits and has been extensively studied for anticarcinogenic effects.Recently, SFN's anti-inflammatory properties through NFK-B and Nrf2 pathways have stoked interest in exploring benefits of SFN for chronic inflammatory disease such as IBD.In vivo studies demonstrate SFN treatment ameliorates DSS colitis in mice.Further, In vitro work shows differential effects of SFN on proliferation and apoptosis of primary vs. transformed cells.To our knowledge, SFN has not yet been studied in human colonoid platform and we sought to examine SFN effects on colonoid viability in comparison to T84 cells, a metastatic colonic carcinoma cell line.Methods: T84 cells and human colonoids were passaged and treated at 48hrs with SFN 0, 5, 10, 20, and 40 uM concentrations.Cellular viability was measured using a commercial resazurin based redox reagent, with fluorescence measurements obtained at 24 and 48hrs.Colonoid measurements were normalized for background fluorescence created by the Matrigel dome.One-way ANOVAs were conducted to compare mean viability scores between treated and untreated T84 cells and colonoids at 24 and 48hrs.Post hoc comparisons were performed using Tukey HSD to examine concentration specific differences.Results: SFN treatment of T84 cells led to a significant effect on viability between groups (F (5, 142) = 10.53P<0.0001)), with increased relative fluorescent scores of 2363 for 10 uM and 2363 at 20 uM SFN vs 1726 in untreated controls (P=0.024and P=0.05, respectively).In contrast, we observed no significant viability differences between untreated controls and the 40uM concentration (p = 0.45).Similarly, in colonoids SFN had a significant effect on mean viability at both 24 and 48 hrs, F (4, 225)=2.474P=0.045.Dose dependent effects were most prominent at 24hrs with mean normalized fluorescence score 158.5 for 20 uM vs 101 for control (p=0.013) and trended towards significance in the 10 uM SFN group with score of 147.6 (p=0.08).However, no significant viability difference was observed between control and 40 uM concentration (p= 0.99).Conclusion: At low and physiologic concentrations of 10 and 20 uM SFN enhances intestinal cell viability in both T84 cells and human colonoids.At concentrations exceeding 40 uM, this proliferative effect is diminished.The mechanisms behind the dose dependent manner of SFN effects warrant further examination to better understand potential strategies for rational use of sulforaphane as a synbiotic therapy for IBD.Further, colonoids appear to be a useful platform for preclinical testing with SFN and may provide a platform to explore cell-specific mechanisms and effects on SFN.