Gum Odina, a novel prebiotic, promotes the expression of sealing tight junction proteins in colon organoids developed from the C57BL/6 mice model

Abstract Maintaining gut-microbiome homeostasis is the biggest issue worldwide as per public health concerns. Gut probiotics not only inhibit pathogen invasion in the systemic circulation but also help us metabolize complex food. Therefore, for decades, gut dysbiosis has been proven to be the gateway to several diseases, leading to comorbidity and even mortality. Prebiotics are natural products, mainly nondigestible food ingredients, that help the selective growth of probiotic bacteria in the gut. This study focuses on the novel Gum-Odina (GO) prebiotic and its efficacy on gut microbial metabolite modulation and maintaining gut barrier integrity. Gut wall enterocytes are integrated by a series of tight junctional (TJ) proteins. This study explains the effect of GO prebiotic-modulated gut metabolites on tight-junctional (TJ) protein expression in a murine colon Organoid model. Fecal microbiota from a colitis patient were used to inoculate the SHIME gut simulator, comprising a colitis control run and a Gum-Odina-supplemented run to enrich commensal bacteria selectively. Metabolites from both groups were then applied to healthy colon organoids. According to the mRNA expression analysis, tight junctional sealing proteins such as Zonula occludens, or ZO-1, Occludin, Claudin-1, 4, and 5 were significantly upregulated in the colon organoids upon Gum-Odina administration, whereas no change in the Junctional Adhesion Molecule-A or JAM-A was observed. Downregulation of sealing TJ proteins is the Hallmark of Leaky gut, which was successfully reversed using the Gum-Odina supplement. Hence, Gum-Odina prebiotics have a promising capability to reduce colitis-induced gut permeability and can be considered to be a therapeutic agent in the future. Competing Interest Statement The authors have declared no competing interest. Footnotes Disclosures: The authors report no competing interests. Abbreviations - IBD - Inflammatory Bowel Disease - C-M - Colitis Metabolites (from SHIME) - GO-M - Gum Odina Metabolites (from SHIME) - C-GO-M - Colitis + Gum Odina Metabolites (treatment group in organoids) - SHIME - Simulator of Human Intestinal Microbial Ecosystem - 3D - Three-dimensional - 2D - Two-dimensional - PBS - Phosphate-Buffered Saline - BSA - Bovine Serum Albumin - DMEM/F-12 - Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12 - PFA - Paraformaldehyde - LPS - Lipopolysaccharide - SEM - Standard Error of the Mean - ANOVA - Analysis of Variance - qPCR - Quantitative (Real-Time) Polymerase Chain Reaction - FITC - Fluorescein Isothiocyanate - GO - Gum Odina - TJ - Tight Junction - ZO-1 - Zonula Occludens-1 - JAM-A - Junctional Adhesion Molecule-A - Cldn-1, -2, -4, -5* - Genes for Claudin-1, Claudin-2, Claudin-4, Claudin-5 - Ocln - Gene for Occludin - Tjp1 - Gene for Tight Junction Protein 1 (ZO-1) - SCFAs - Short-Chain Fatty Acids - DSS - Dextran Sodium Sulfate - GAPDH - Glyceraldehyde 3-Phosphate Dehydrogenase (housekeeping gene) - TNF-α - Tumor Necrosis Factor-alpha - IL-6 - Interleukin-6 - IFN-γ - Interferon-gamma - IL-12 - Interleukin-12 - IL-10 - Interleukin-10