ENAapplication/xmlftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR177/007/SRR1777617/SRR1777617_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR177/006/SRR1777616/SRR1777616_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR177/004/SRR1777614/SRR1777614_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR177/007/SRR1777617/SRR1777617_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR177/003/SRR1777613/SRR1777613_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR177/004/SRR1777614/SRR1777614_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR177/005/SRR1777615/SRR1777615_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR177/002/SRR1777612/SRR1777612_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR177/006/SRR1777616/SRR1777616_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR177/002/SRR1777612/SRR1777612_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR177/003/SRR1777613/SRR1777613_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR177/005/SRR1777615/SRR1777615_1.fastq.gzprimaryOK200GenomicsPI: D. Brent Polk, The Saban Research Institute, Children's Hospital Los Angeleshttps://www.ebi.ac.uk/ena/browser/view/PRJNA273889Mus musculusxref:PubMed:26072395PURPOSE: The goal of this study was to determine the gene expression networks regulated by tumor necrosis factor receptor 2 (TNFR2, or Tnfrsf1b) and to evaluate their potential bearing on immune cell subsets and inflammatory bowel disease (IBD). METHODS: mRNA-seq was performed on isolated distal colons from TNFR2-knockout and wildtype mice. Differentially expressed transcripts were compared to human ulcerative colitis microarray datasets on Gene Expression Omnibus and to mouse immunological expression datasets at the Immunological Genome Project. RESULTS: We identified 252 mouse transcripts whose expressions were significantly altered by the loss of TNFR2. The majority of these transcripts (228 of 252, ~90%) were downregulated in TNFR2-/- colons. TNFR2-regulated genes were able to positively discriminate between ulcerative colitis patients and healthy individuals with ~80% accuracy. Many TNFR2-regulated genes were also highly expressed in CD8+ T cells. CONCLUSIONS: Downregulation of TNFR2 is associated with a gene expression profile that is prominent in IBD and supportive of the role of CD8+ T cells in IBD pathogenesis. MANUSCRIPT ABSTRACT: Increased tumor necrosis factor (TNF) production has been associated with inflammatory bowel disease (IBD), and anti-TNF therapy is a common therapeutic for this patient population. However, the role of TNF or its receptors (TNFR1 and TNFR2) in the immunopathogenesis of inflammatory bowel disease (IBD) remains unclear. Here we report that TNFR2 is protective in spontaneous (IL-10 knockout) and chemically (azoxymethane/dextran sodium sulfate)-induced mouse models of colitis and colitis-associated cancer. Mechanistically, TNFR2-deficiency in hematopoietic cells significantly increased incidence and severity of colitis and colitis-associated cancer characterized by a selective expansion of CD8+ T cells. We identified TNFR2-regulated genes in the colon that were specific for CD8+ T cells, interacted with multiple IBD risk genes, and are important regulators of CD8+ T cell biology. TNFR2 regulated CD8+ T-cell-specific genes that act as genetic susceptibility modifiers for IBD to mitigate the development of a pro-colitogenic milieu. Antibody-mediated depletion of CD8+ T cells prevented colonic inflammation and significantly reduced pathology in IL10-/-/TNFR2-/- deficient mice. Furthermore, adoptive transfer of TNFR2-/- naïve CD8+ T cells resulted in more severe disease than with wildtype naïve CD8+ T cells. Our findings provide insight into the disease modifier role of TNFR2 in the immunopathogenesis of IBD through the modulation of CD8+ T cell responses and support future investigation of this therapeutic target, especially in the subset of IBD patients with CD8+ T-cell dysfunction. Overall design: Total RNA from distal colons of 8 week-old male wildtype C57Bl/6 and TNFR2-/- mice (n=3 each) was isolated using the PureLink RNA kit (Ambion, Life Technologies). RNA samples were submitted to the Genomic Services Lab at the HudsonAlpha Institute for Biotechnology (Huntsville, AL) for multiplex library preparation, mRNA enrichment, and sequencing. Sequencing was performed to an average depth of 50M paired-end 50bp reads per sample (HiSeq, Illumina, San Diego, CA). Data files containing raw reads were aligned to the mouse genome using Tophat2/Bowtie2. Alignments were assembled into transcript representations with Cufflinks, and statistical tests for differential expression were performed with Cuffdiff 2. An adjusted P value < 0.05 (q<0.05) from the Cuffdiff 2 output was used as the cutoff for statistical significance.ENATNF-R-II, Tumor Necrosis Factor Receptor Superfamily, TNFRSF1B Receptor, TNFR p80, CD120b Antigen, Tumor Necrosis Factor Receptor 2, Transcriptome, 120b Antigen, determination, Transcriptome Profile, Laboratory, Mus domesticus, Expression Profiles, mouse, Receptors, Gene Expression Profile, Gene, Profiles, TNFBR, Appendix Epiploica, Tumor Necrosis Factor Receptor Type II, House Mouse, hindgut, Tnfr-2, mice C57BL/6xCBA/CaJ hybrid, Tnfr-1, Mus muscaris, whole transcriptome, Antigens, Gene Expression, TNF-RII, TNF-alphaR2, TNFRII, Member 1B, large bowel, House, Mus, TNFRSF1B, Taenia Coli, Expression Signatures, Gene Expression Signatures, chemical analysis, Appendix, Antigen, Mus musculus domesticus, p75TNFR, TNFR1B, Gene Expression Signature, Omental., Expression Profile, Mice, Omental Appendices, Tumor Necrosis Factor, Transcriptome Profiles, Mus musculus, TNF-R2, Omental Appendix, Swiss, mice, Profile, p75, Swiss Mouse, posterior intestine, Appendices, TNF-sR75, House Mice, Swiss Mice, Tumor Necrosis Factor Receptor 75, CD 120b, Omental, Tumor Necrosis Factor Receptor Type 2, Signatures, CD120b, CD 120b Antigen, Tnfr2, domesticus, Laboratory Mice, CD, TNFalpha-R2, TNFR2, Expression Signature, COLON, Gene Expression Profiles, Transcriptomes, Mouse, assay, Receptor, house mouse, Signature, TNF-R75, TNFR p75, Laboratory Mouse, CD120b Antigens, TBPII, TNFR80Mus musculus, Laboratory Mice., House, Mus, Laboratory, Swiss, Mus domesticus, mouse, Mus musculus domesticus, Swiss Mouse, mouse <Mus musculus>, Mouse, House Mice, Swiss Mice, house mouse, Mice, Laboratory Mouse, House Mouse, mice C57BL/6xCBA/CaJ hybrid, domesticus, Mus muscarisfalseMus musculusTranscriptome analysis of TNFR2-knockout mouse colon2025-09-242015-08-18PRJNA273889GSE654081009026072395