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Next Generation Sequencing Facilitates Quantitative Analysis of Il17ra villin cre -/+ colon transcriptomes at day 10 post C. rodentium infection

ABSTRACT: We report analysis of colonic transcriptome in gut epithelial cells specific deficient mouse (Il17ra villin cre +) and littermate control mouse (Il17ra villin cre -) infected with Citrobater rodentium (C. rodentium) for 10 days. We found RNA-seq data show Nox family genes and defensin genes are downregulated in Il17ra villin cre + mice. It is interestingly Tnfsf13 was also downregulated in Il17ra villin cre + mice. These genes expressions were confirmed by qRT–PCR validation was performed using TaqMan assays. Finally, this study provides first detailed analysis of C. rodentium infected colonic transctiptome in Il17ra villin cre + mice, with biologic replicates, generated by RNA-seq technology.

ORGANISM(S): Mus musculus

PROVIDER: GSE160157 | GEO | 2020/12/08

REPOSITORIES: GEO

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Project description:Opioids analgesics are frequently prescribed in the United States and worldwide. However, serious side effects such as addiction, immunosuppression and gastrointestinal symptoms limit their use. It has been recently demonstrated that morphine treatment results in significant disruption in gut barrier function leading to increased translocation of gut commensal bacteria. Further study indicated distinct alterations in the gut microbiome and metabolome following morphine treatment, contributing to the negative consequences associated with opioid use. However, it is unclear how opioids modulate gut homeostasis in the context of a hospital acquired bacterial infection. In the current study, a mouse model of C. rodentium infection was used to investigate the role of morphine in the modulation of gut homeostasis in the context of a hospital acquired bacterial infection. Citrobacter rodentium is a natural mouse pathogen that models intestinal infection by enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC) and causes attaching and effacing lesions and colonic hyperplasia. Morphine treatment resulted in 1) the promotion of C. rodentium systemic dissemination, 2) increase in virulence factors expression with C. rodentium colonization in intestinal contents, 3) altered gut microbiome, 4) damaged integrity of gut epithelial barrier function, 5) inhibition of C. rodentium-induced increase in goblet cells, and 6) dysregulated IL-17A immune response. This is the first study to demonstrate that morphine promotes pathogen dissemination in the context of intestinal C. rodentium infection, indicating morphine modulates virulence factor-mediated adhesion of pathogenic bacteria and induces disruption of mucosal host defense during C. rodentium intestinal infection in mice. This study demonstrates and further validates a positive correlation between opioid drug use/abuse and increased risk of infections, suggesting over-prescription of opioids may increase the risk in the emergence of pathogenic strains and should be used cautiously. Therapeutics directed at maintaining gut homeostasis during opioid use may reduce the comorbidities associated with opioid use for pain management.

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Next Generation Sequencing Facilitates Quantitative Analysis of Il17ra villin cre -/+ colon transcriptomes at day 10 post C. rodentium infection

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