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ABSTRACT: Paneth cells, the Microbiome, and Crohn’s Disease Susceptibility

PROVIDER: PRJNA317429 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5762490_1.fastq.gz	Fastqsanger.gz
	SRR5762490_2.fastq.gz	Fastqsanger.gz
	SRR5762500_1.fastq.gz	Fastqsanger.gz
	SRR5762500_2.fastq.gz	Fastqsanger.gz
	SRR5762510_1.fastq.gz	Fastqsanger.gz

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Gene expression analysis of crypt base epithelial cells from WT and Nod2-/- mice

Project description:Nod2 has been extensively characterized as a bacterial sensor that induces an antimicrobial and inflammatory gene expression program. Therefore, it is unclear why Nod2 mutations that disrupt bacterial recognition are paradoxically among the highest risk factors for Crohn’s disease, which involves an exaggerated immune response directed at intestinal bacteria. Previous studies from our lab have shown that mice deficient in Atg16L1, another Crohns disease susceptibility gene, develop abnormalities in Paneth cells, specialized epithelial cells in the small intestine involved in antimicrobial responses. The goal of our study is to determine whether Nod2 deficiency leads to differences in the transcriptional profile of Paneth cells, ultimately leading to small intestinal inflammation. Small intestinal sections (ileum) of 8 week old WT and Nod2-/- mice were fixed in methacarn and embedded in paraffin. The Leica LMD6000 Laser Microdissection system was used to capture crypt base epithelial cells to enrich for Paneth cells. RNA was extracted from these cells, followed by cDNA synthesis and qPCR to confirm enrichment of Paneth cells using unique markers (a-defensins).

2014-07-01 | E-GEOD-58948 | biostudies-arrayexpress

Genome-wide association study of abnormal Paneth cell phenotype in Japanese patients with Crohn's disease

Project description:Morphological patterns of Paneth cells are a cellular biomarker in western Crohn’s disease (CD) patients. They integrate genetics and environmental factors, and are associated with outcomes. To broaden the applications of Paneth cell phenotype, identification of novel genetic determinants and clinical validation in other ethnic groups is critical.

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Gene expression analysis of crypt base epithelial cells from WT and Nod2-/- mice

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Project description:We previously showed that abnormal morphology phenotype of ileal Paneth cells (Paneth cell phenotype [PCP]; as a surrogate for PC function) correlate with genetics, microbiota compositions, and aggressive outcome in Crohn’s disease (CD) patients. Given the shared genetics and clinical features between CD and ulcerative colitis (UC), we hypothesized that abnormal PCP also negatively modulates UC outcomes. As PCs has the highest density in the ileum, we further hypothesized that abnormal PCP from the terminal ileum could increase the risk of development of pouch complications after UC total colectomy and ileal pouch anal anastomosis (IPAA).

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16S rRNA analysis from wild type mice that were either fed or not with western diet for 3 generations

Project description:We found that western diet consumption resulted in decrease in the percentage of normal Paneth cell population in wild type mice, indicating that western diet could negatively affect Paneth cell function. Subsequent generations of western diet consumption further reduced percentages of normal Paneth cell population. We performed fecal microbiota composition profiling. Male mice were used at 4-5 weeks of age. Fecal samples were collected for microbiome analysis.

2023-01-01 | E-MTAB-10995 | biostudies-arrayexpress

16SrRNA analysis from wildtype mice that were either fed or not with low protein diets for 4 weeks

Project description:We found that low protein diet consumption resulted in decrease in the percentage of normal Paneth cell population in wild type mice, indicating that low protein diet could negatively affect Paneth cell function. We performed fecal microbiota composition profiling. Male mice were used at 4-5 weeks of age. Fecal samples were collected for microbiome analysis.

2023-07-02 | E-MTAB-11277 | biostudies-arrayexpress

Project description:Paneth Cell Defects in Crohn’s Disease Patients Promote Dysbiosis

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transcriptome of purified PC and non-PC fractions: GR-WT & GR-KO

Project description:Paneth Cells (PCs) are secretory cells located in the crypts of Lieberkühn of the small intestine. They produce antimicrobial peptides (AMPs) and are thought to keep the microbiome under control. The cytokine TNF, which functions mainly via its TNF receptor 1 (TNFR1 or P55), is a driving force in many inflammatory bowel disease (IBD) patients, but its impact on PCs is poorly known. We have generated intestinal glucocorticoid receptor knockout mice (GR villin KO) and investigated the diffrences between WT and KO paneth and non-paneth cell fractionf from the cell sorting.

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Targeted transcriptome analysis of isolated intestinal epithelial cells from mice with constitutive Ptpn2-deficiency

Project description:Background & aims: Loss-of-function variants in the PTPN2 gene are associated with increased risk of inflammatory bowel disease. We recently showed that Ptpn2 is critical for intestinal epithelial cell (IEC) barrier maintenance, IEC-macrophage communication, and modulation of the gut microbiome in mice, restricting expansion of a small intestinal pathobiont associated with inflammatory bowel disease. Here, we aimed to identify how Ptpn2 loss affects ileal IEC subtypes and their function in vivo. Methods: Constitutive Ptpn2 wild-type, heterozygous, and knockout (KO) mice, as well as mice with inducible deletion of Ptpn2 in IECs, were used in the study. Investigation was performed using imaging techniques, flow cytometry, enteroid culture, and analysis of gene and protein levels of IEC markers. Results: Partial transcriptome analysis showed that expression of Paneth cell-associated antimicrobial peptides Lyz1, Pla2g2a, and Defa6 was down-regulated markedly in Ptpn2-KO mice compared with wild-type and heterozygous. In parallel, Paneth cell numbers were reduced, their endoplasmic reticulum architecture was disrupted, and the endoplasmic reticulum stress protein, C/EBP-homologous protein (CHOP), was increased in Ptpn2-KO mice. Despite reduced Paneth cell number, flow cytometry showed increased expression of the Paneth cell-stimulatory cytokines interleukin 22 and interferon γ+ in CD4+ T cells isolated from Ptpn2-KO ileum. Key findings in constitutive Ptpn2-KO mice were confirmed in epithelium-specific Ptpn2ΔIEC mice, which also showed impaired lysozyme protein levels in Paneth cells compared with Ptpn2fl/fl control mice. Conclusions: Constitutive Ptpn2 deficiency affects Paneth cell viability and compromises Paneth cell-specific antimicrobial peptide production. The observed effects may contribute to the increased susceptibility to intestinal infection and dysbiosis in these mice.

2023-04-10 | GSE181923 | GEO

Single cell RNA sequencing analysis of germ free (GF) mouse Paneth cells and microbiome-transferred exGF mouse Paneth cells.

Project description:The single cell level Paneth cell transcriptome under gnotobiotic condition has not been resolved. How gut microbiome modulates Paneth cell transcriptome at single cell level in germ-free mice was not known. We used a flow cytometry method to isolate highly pure ileal Paneth cells from germ free (GF) Paneth cell reporter mice (Lyz1-3'UTR-IRES-CreER) and from exGF Paneth cell reporter mice that were transplanted with wild type C57B/l6 mouse gut microbiota (exGF+B6M). These isolated Paneth cells were subjected to single cell RNA sequencing by 10xGenomics.

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