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Mucosal Glycan Foraging Enhances the Fitness and Transmission of a Saccharolytic Human Distal Gut Symbiont: ECF mutant

ABSTRACT: Growth of isogenic parent ('tdk strain'), 5xECF-sigma factor mutant and complemented mutant strain in the cecum of NMRI inbred mice fed a glycan-restricted, 'simple sugar' diet (35% each glucose and sucrose). Bacteria were collected from mouse distal gut 10 days after colonization.

ORGANISM(S): Bacteroides thetaiotaomicron Bifidobacterium longum

PROVIDER: GSE11953 | GEO | 2008/11/21

SECONDARY ACCESSION(S): PRJNA109257

REPOSITORIES: GEO

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Project description:Dr. Lebrilla's lab is working on another set of transcriptomics experiments which will measure whether HMO (human milk oligosacharides) elicit a differential response on Caco2 in the presence of B.infantis. They wish to supplement the data obtained with the Illumina platform with glycan-related expression data obtained with the Gene-chips to achieve a more complete understanding on how Caco2 cells respond to the presence of HMOs and B.infantis. The transcriptomics data is being complemented by qPCR based bacterial-epithelial cells binding assays, and glycan arrays assays for which they have initiated a collaboration with the CFG Core H (David Smith). The influence, signaling and adhesion between Bifidobacterium infantis, a probiotic microorganism abundant in infant feces, and Caco2 epithelial cells. Our hypothesis is that HMOs act as signaling molecules for both Caco2 cells and B. infantis to indicate the presence in the gut, of a nutrient niche adapted for the nourishment and recruitment of HMO-consuming probiotic microorganism. HMOs therefore could play a role in the expression and binding to GBPs both on epithelial cells and in probiotic gut microorganisms. Preliminary transcriptomics data analysis obtained by using the Illumina Human Ref-8 Expression Bead Chip, has revealed that co-incubation of Caco2 cells with HMO upregulates genes belonging to chemokines, adhesion molecules, growth factors and glycan degradation, thus indicating that HMOs are actively recognized by these cells. We are working on another set of transcriptomics experiments which will measure whether HMO elicit a differential response on Caco2 in the presence of B.infantis. We wish to supplement the data obtained with the Illumina platform with glycan-related expression data obtained with the Gene-chips to achieve a more complete understanding on how Caco2 cells respond to the presence of HMOs and B.infantis. The transcriptomics data is being complemented by qPCR based bacterial-epithelial cells binding assays, and glycan arrays assays for which we have initiated a collaboration with the CFG Core H (David Smith). We are requesting the analysis of 16 RNA samples from Caco2 cells on the Glyco-gene chips, for testing respectively 4 independent biological replicates for each of the 4 different co-incubation conditions. RNA preparations from Caco2 cells were sent to Microarray Core (E). The RNA was amplified, labeled, and hybridized to GLYCO_v3 microarrays.

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Mucosal Glycan Foraging Enhances the Fitness and Transmission of a Saccharolytic Human Distal Gut Symbiont: ECF mutant

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