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Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and RNF138-/- mice colorectal transcriptomes in DSS, AOM/DSS and organoids samples

ABSTRACT: Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to analysis NGS-derived transcriptome profiling (RNA-seq) in DSS induced chronic inflammation, AOM/DSS induced colitis-associated colorectal tumorigenesis and organoids isolation from colitis-associated colorectal tumorigenesis Methods: DSS, AOM/DSS and organoids mRNA profiles of wild-type (WT) and RING Finger 3 (RNF138−/−) mice were generated by deep sequencing, in triplicate, using Illumina HiSeq 2500. The sequence reads were trimmed for low-quality sequence, then mapped to mm10 whole genome using STAR v2.6.1d Results: Using an optimized data analysis workflow, the padj <0.05 and fold change >2 were refered as differential expression. There are 987, 2649 and 2373 differential genes were found in RNF138-/- compared with Wild Type in DSS, AOM/DSS and organoids, respectively Conclusions: Our study revealed NFκB pathway is the main activation pathway regulated by RNF138 loss

ORGANISM(S): Mus musculus

PROVIDER: GSE144051 | GEO | 2022/04/30

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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Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and RNF138-/- mice colorectal transcriptomes in DSS, AOM/DSS and organoids samples

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