Project description:Cryptosporidiosis is a zoonotic disease caused by infection with the oocyst of Cryptosporidium in human and animals. MicroRNA (miRNA) emerges as important player in regulating the innate immune response against parasitic infection. Here, we compared miRNA profiles of the glandular stomach of Cryptosporidium muris (C. muris) infected and un-infected BALB/c mice using microarray sequencing. A total of 10 miRNAs (including 3 upregulated and 7 downregulated miRNAs) with significant expression differences (|FC| ≥ 2 and P value test < 0.05) were screened after C. muris infected the glandular stomach of BALB/c mice for 8 hours. MiRWalk and miRDB online bioinformatics software were used to predict the target genes of differentially expressed miRNAs. Gene Ontology (GO) and KEGG enrichment analyses were performed for annotate the target genes. GO terms indicates that many are associated with the relevant generic transcription and ion transport. In addition, the KEGG analyses showed that the target genes were strictly related to a diverse types of tumor disease progression and the antipathogen immunity pathway. In the current study, we first reported the changes of miRNA expression profile in glandular stomach of BALB/c mice at the early phase of C. muris invasion. As such, dysregulation of miRNA expression profile may contribute to our understanding of the Cryptosporidiosis pathology. The result reported in this paper provide a new perspective into the miRNA regulatory mechanisms of Cryptosporidiosis, which may help to develop effective control strategies against Cryptosporidium.
Project description:The goal of this project was to compare the metabolite profiles of the: mouse gastric antrum and the mouse gastric corpus, the mouse gastric antrum and the mouse gastric antrum isolated glands, and the mouse gastric corpus and the mouse gastric corpus isolated glands.
Project description:We generated a novel Six2-Cre+/-PKAcaRfl/wt (CA-PKA) CA-PKA mouse in which expression of constitutive-active PKAcaR was induced in gastric mesenchyme progenitors. CA-PKA mice showed disruption of gastric homeostasis characterized by aberrant mucosal development and epithelial hyperproliferation; ultimately developing multiple features of gastric corpus preneoplasia including decreased parietal cells, mucous cell hyperplasia, spasmolytic peptide expressing metaplasia (SPEM) with intestinal characteristics and dysplastic and invasive cystic glands. Our results show that constitutively active PKAcaR in the stomach mesenchyme nonautonomously disrupts gastric homeostasis characterized by increased epithelial proliferation and aberrant epithelial maldevelopment, ultimately leading to gastric preneoplasia.
Project description:We explored the potential of human gastric organoids as models for mucus production. Immunofluorescence staining confirmed that the organoids produced mucus containing MUC5AC and MUC6. The luminal mucus had viscoelastic properties similar to those of native human gastric mucus, as determined by particle tracking microrheology. To collect organoid-produced gastric mucus, termed bioengineered gastric mucus (BGM), organoids were cultured as monolayers at the air-liquid interface (ALI), and apically-secreted mucus was harvested and analyzed by MUC5AC ELISA, proteomics, CryoFE-SEM, and bulk rheometry. BGM contained high-molecular weight molecules also found in native gastric mucus, including MUC5AC. Proteomic analysis confirmed that BGM contained MUC5AC, MUC6, MUC1, and other stomach-specific molecules such as gastricsin, olfactomedin 4, and gastrokine. CryoFE-SEM showed that both BGM and native mucus had a porous structure and a characteristic honeycomb scaffold. Bulk rheometry confirmed that BGM exhibited shear thinning and predominantly elastic behavior, consistent with native mucus. Collectively, these findings indicate that BGM is an accessible alternative to native gastric mucus that can be produced on-demand for in vitro studies.
Project description:Purpose: Sox2 expression marks gastric stem and progenitor cells, raising important questions regarding the genes regulated by Sox2 and the role of Sox2 itself during stomach homeostasis and disease. The goal of this study is to determine the function of and the genes regulated by Sox2 in the stomach. Methods: Sox2 ChIP-enriched DNA and input DNA was isolated from gastric glands of adult antrum from Sox2 KO and Sox2 WT mice. DNA was purified and genomic libraries were prepared as described (Sulahian et al., 2014), using four micrograms of goat anti-SOX2 (AF2018, R&D). Libraries were sequenced (50 bp, single-end reads) on an Illumina Hi-Seq 2000 instrument. Results: Sox2 is dispensiable for gastric stem cell self-renewal and epithelial homeostasis, however modulates the expression of wnt, intestinal and cancer related genes