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Single cell RNAseq of mouse colon stroma during chronic DSS and homeostatic condition

ABSTRACT: We implemented a murine model of intestinal inflammation based on oral administration of multiple cycles of low dose dextran sulfate sodium (DSS) to induce epithelial injury and ECM deposition. Mice were subjected to 3 repetitive cycles of DSS displayed progressive accumulation of immune cell infiltrates associated with excessive deposition of collagen fibers. Lamina propria cells from water-fed and DSS-fed mice were isolated using enzymatic digestion, and enriched for stromal cells by FACS using antibodies excluding hematopoietic cells (CD45), epithelial cells (EpCAM), and erythrocytes. Prepared single cell suspensions were then profiled using the 10x Chromium V2 droplet-based single cell RNA sequencing platform. Tissues were collected from proximal and distal colon.

ORGANISM(S): Mus musculus

PROVIDER: GSE172261 | GEO | 2022/01/10

REPOSITORIES: GEO

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Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are using transcriptome profiling (RNA-seq) to evaluate the effects of anti-S100a9 antibody on the global transcriptome of the colon tissues of the AOM/DSS mouse model (a model that mimics the human colitis-associated colon cancer development). Methods: 36 five-week-old male ICR mice were randomized divided into three groups: control (i.e. no AOM/DSS and antibody treatment), AOM/DSS+IgG Ab (1.5 mg/kg), and AOM/DSS+anti-S100a9 Ab (1.5 mg/kg). Mice were intraperitoneal injected with a single dose of 10 mg/kg azoxymethane (AOM) (A5486; Sigma) on day 1. One week after the AOM injection, mice were given three cycles of DSS (cycle 1: 2%, 7 days; cycle 2: 1.5%, 5 days; and cycle 3: 1.5%, 5 days, DSS: 36–50 kDa; MP Biomedicals, CA, USA) in their drinking water, and then distilled water until the end of the experiment. Antibodies were administrated intravenously every two days during the three cycles of DSS treatment. Mice were sequentially killed randomly at the end of the 18th week, and at least five mice were killed for each group at each time point. RNAs were extracted by Trizol and sequenced by Solexa high-throughput sequencing service (Oebiotech, Shanghai, China). Data were extracted and normalized according to the manufacturer’s standard protocol.Each group has three mices' colon tissues be tested. Results: Log-fold changes of up- or down-regulated mRNAs between the control and experiment group were selected with a significance threshold of p<0.05. There are 1017 mRNAs were up-regulated and 815 were down-regulated in “AOM/DSS+IgG Ab" group comparing to “control" group. There are 385 mRNAs were up-regulated and 163 were down-regulated in “AOM/DSS+anti-S100a9 Ab" group comparing to “control" group. There are 1314 mRNAs were up-regulated and 968 were down-regulated in “AOM/DSS+anti-S100a9 Ab" group comparing to “AOM/DSS+IgG Ab". Conclusions: Our study describes the global transciptome changes of colon tissues of the AOM/DSS mouse model induced by anti-S100a9 antibody treatment.

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Single cell RNAseq of mouse colon stroma during chronic DSS and homeostatic condition

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