Journal: Gut

Article Title: Parasutterella excrementihominis exacerbates experimental colitis and colitis-associated colorectal cancer via pathogenic NETosis activation

doi: 10.1136/gutjnl-2025-335887

Figure Lengend Snippet: Elevated abundance of P. excrementihominis in UC patients.( A–C ) Relative abundance of P. excrementihominis in IBD faecal samples using available metagenomic data from the GMrepo database: Cardiff cohort (PRJEB7949; UK) and PRISM/STINKI cohorts (PRJNA385949; USA: Atlanta); Cardiff (n=12 (Health) or 20 (active IBD)); and PRISM/STINKI (n=10 (Health) or 10 (active IBD)). ( D ) Detection of P. excrementihominis in the faeces of 115 UC patients (36 in remission and 79 with active disease) and 20 healthy controls using quantitative PCR. ( E ) Correlation between P. excrementihominis levels and the Mayo scores in UC, the C-reactive protein values in UC and the erythrocyte sedimentation rate (ESR) values in UC were calculated by Spearman correlation analyses. ( F ) Detection of P. excrementihominis in the faeces of healthy controls (n=20), UC patients in UC-disease duration (<5, n=40), UC-disease duration (5–15, n=58) and UC-disease duration (>15, n=17) using quantitative PCR. ( G ) Detection of P. excrementihominis in the faeces of healthy controls (n=20), UC patients without CRC development (n=109) and CAC patients (n=6) using quantitative PCR. ( H ) Detection of P. excrementihominis in the inflamed and normal (non-inflamed) colonic mucosa from active UC patients (n=34). ( I–J ) Detection of P. excrementihominis in the inflamed biopsies from active UC (n=25) and the colonic mucosa from UC in remission (n=20) by FISH. (The data are presented as mean±SEM in each group. Statistical analysis was performed by one-way analysis of variance, unpaired Student’s t test or paired Student’s t test. NS, non-significant; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.) CAC, colitis-associated colorectal cancer; CRC, colorectal cancer; IBD, inflammatory bowel disease; LDA, linear discriminant analysis; UC, ulcerative colitis.

Article Snippet: Each reaction was performed in triplicate with 2X SYBR Green Premix Pro Taq HS qPCR Tracking Kit (ROX2 plus, AG11735, Accurate Biotechnology (Hunan) Co., Ltd), primers and template DNA.

Techniques: Real-time Polymerase Chain Reaction, Sedimentation

Journal: Gut

Article Title: Parasutterella excrementihominis exacerbates experimental colitis and colitis-associated colorectal cancer via pathogenic NETosis activation

doi: 10.1136/gutjnl-2025-335887

Figure Lengend Snippet: P. excrementihominis ( P.e ) exacerbates DSS-induced colitis in mice. ( A ) Mice gavaged with live P.e , heat-killed P.e ( P.e-HK ), bacterial culture supernatant ( P.e-CS ) or control medium were subjected to DSS-induced experimental colitis. ( B–D ) In the P.e group, luminal stool samples, colonic tissues and small intestinal tissues were collected from mice at the indicated time points. Then, the abundance of P.e . was assessed using quantitative PCR analysis. ( E ) Assessment of body weight changes. ( F ) Disease activity index was monitored after DSS treatment. ( G–H ) Mice were euthanised on day 7 after DSS treatment, and colon lengths were measured. ( I–J ) Colon sections were examined histologically and lesion areas were quantified (scale bars: 100 µm). ( K–L ) Immunostaining was performed to analyse the infiltration of colonic Ly6G + neutrophils, F4/80 + monocyte macrophages and CD8 + T cells (scale bars: 100 µm). (The data are presented as mean±SEM, n=6 mice per group, and statistical analysis was performed by one-way analysis of variance (ANOVA). DSS, dextran sulphate sodium; NS, non-significant. *p<0.05, **p<0.01, ***p<0.001. Data shown are representative of three independent experiments.

Article Snippet: Each reaction was performed in triplicate with 2X SYBR Green Premix Pro Taq HS qPCR Tracking Kit (ROX2 plus, AG11735, Accurate Biotechnology (Hunan) Co., Ltd), primers and template DNA.

Techniques: Control, Real-time Polymerase Chain Reaction, Activity Assay, Immunostaining

Journal: Gut

Article Title: Parasutterella excrementihominis exacerbates experimental colitis and colitis-associated colorectal cancer via pathogenic NETosis activation

doi: 10.1136/gutjnl-2025-335887

Figure Lengend Snippet: P. excrementihominis ( P.e ) triggers NETosis to intensify colitis.( A ) Colon tissues from control mice or DSS mice gavaged with live P.e and control medium (n=3) were harvested for RNA-sequencing (RNA-seq). ( B ) Histogram depicting the quantity of differentially expressed genes in the transcriptome, was analysed by RNA-seq (upregulated genes are indicated in grey, and downregulated genes are indicated in red). ( C ) Volcano plot illustrating differential gene expression between two groups of mice, was visualised by log2 fold change, with high expressed genes indicated in purple and low expressed genes in red. (When log2FC>0, the gene is upregulated; when log2FC<0, it indicates that the gene is downregulated.) ( D–F ) Heatmap displaying differential gene expression between two groups of mice, was visualised by log2 fold change, with high expressed genes indicated in red and low expressed genes in dark purple. (When log2FC >0, the gene is upregulated; When log2FC <0, it indicates that the gene is downregulated. ( G ) KEGG pathway enrichment analyses of the differential genes in P. e -colonised mice compared with control mice after exposure to DSS treatment. ( H ) Relative mRNA expression levels of cytokines, including S100A8/9, MMP8/9, LCN2, CXCR2, CTSG, CSF3, CSF3R and PADI4, were determined by real-time qPCR and normalised to 18S. ( I–J ) Immunofluorescence double staining of MPO (green) and CitH3 (red) to detect NETs formation (scale bars: 50 µm). ( K–P ) The pathogenic effect of P.e was studied in the PADI4-/- and their littermate wild-type (WT) mice treated with DSS. ( K ) Assessment of body weight changes. ( L ) Disease activity index was monitored after DSS treatment. ( M–N ) Mice were euthanised on day 7 after DSS treatment, and colon lengths were measured. ( O–P ) Colon sections were examined histologically and lesion areas were quantified (scale bars: 100 µm). (The data are presented as mean±SEM, n=6 mice per group, and statistical analysis was performed by one-way analysis of variance (ANOVA). NS, non-significant; *p<0.05; **p<0.01; ***p<0.001. Data shown are representative of three independent experiments.) CitH3, citrullinated histone H3; DAI, Disease Activity Index; DSS, dextran sulfate sodium; ECM, extracellular matrix; H&E, haematoxylin and eosin staining; IL, Interleukin; JAK, janus kinase; KEGG, Kyoto Encyclopedia of Genes and Genomes; MPO, myeloperoxidase; NETs, neutrophil extracellular traps; STAT, signal transducers and activators of transcription; TGF, transforming growth factor; TNF, tumour necrosis factor; WT, wild type.

Article Snippet: Each reaction was performed in triplicate with 2X SYBR Green Premix Pro Taq HS qPCR Tracking Kit (ROX2 plus, AG11735, Accurate Biotechnology (Hunan) Co., Ltd), primers and template DNA.

Techniques: Control, RNA Sequencing, Gene Expression, Expressing, Immunofluorescence, Double Staining, Activity Assay, Staining

Journal: Gut

Article Title: Parasutterella excrementihominis exacerbates experimental colitis and colitis-associated colorectal cancer via pathogenic NETosis activation

doi: 10.1136/gutjnl-2025-335887

Figure Lengend Snippet: P. excrementihominis ( P.e ) promotes colitis-associated tumourigenesis. ( A–F ) Male C57/BL6 mice administered with live P.e, bacterial culture supernatant ( P.e- CS) or control medium (Ctrl) were subsequently subjected to AOM/DSS treatment to induce CAC modelling. ( A ) Body weight changes were monitored after AOM/DSS administration per 3 days. ( B ) Mice were euthanised on day 61, and colon lengths were measured. ( C ) Tumour volumes and numbers were measured. ( D ) Representative images of the colons from the indicated group of mice. ( E–F ) Colon sections were examined histologically and lesion areas were quantified (scale bars: 100 µm). ( G ) Representative IHC images of Ki67 staining in colon tumours (scale bars: 100 µm). ( H ) Representative immunofluorescence images showing MPO (green) and CitH3 (red) staining in colonic tissues of mice (scale bars: 100 µm). ( I ) Quantitative analyses of Ki67 and MPO + CitH3 + positive cells in colonic tissues of CAC mice. ( J ) Relative mRNA expression levels of cytokines including MPO, S100A8/9, MMP3/9/10, CCL4, LTF, CAMP and CTSG were determined by real-time qPCR and normalised to 18S. (The data are presented as mean±SEM in each group. n=4–10. NS, non-significant; *p<0.05; **p<0.01, ***p<0.001, ****p<0.0001 by unpaired Student’s t test. Data shown are representative of three independent experiments.) AOM, azoxymethane; CAC, colitis-associated colorectal cancer; CitH3, citrullinated histone H3; DSS, dextran sulphate sodium; MPO, myeloperoxidase.

Article Snippet: Each reaction was performed in triplicate with 2X SYBR Green Premix Pro Taq HS qPCR Tracking Kit (ROX2 plus, AG11735, Accurate Biotechnology (Hunan) Co., Ltd), primers and template DNA.

Techniques: Control, Staining, Immunofluorescence, Expressing

Journal: Gut

Article Title: Parasutterella excrementihominis exacerbates experimental colitis and colitis-associated colorectal cancer via pathogenic NETosis activation

doi: 10.1136/gutjnl-2025-335887

Figure Lengend Snippet: GSDMD deletion mitigates metabolite-driven CAC progression. ( A ) GSDMD flf l mice and GSDMD flf l S100A8-Cre mice received oral administration of either 6-HHA, Suc or PBS via gavage underwent AOM/DSS treatment to induce CAC. ( B ) Body weight changes were monitored after AOM/DSS administration per 3 days. ( C ) Tumour volumes and numbers were measured. ( D ) Mice were euthanised on day 61, and colon lengths were measured. ( E ) Representative images of the colons from the indicated group of mice. ( F–G ) Colon sections were examined histologically, and the lesion areas were quantified (scale bars: 200 µm). ( H ) Relative mRNA expression levels of MPO, S100A8/9 and MMP9/10 were determined by real-time qPCR and normalised to 18S. (The data are presented as mean±SEM in each group. n=4–6. Statistical analysis was performed by one-way analysis of variance (ANOVA). NS, non-significant; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Data shown are representative of three independent experiments.) AOM, azoxymethane; CAC, colitis-associated colorectal cancer; DSS, dextran sulphate sodium; GSDMD, gasdermin D; MPO, myeloperoxidase.

Article Snippet: Each reaction was performed in triplicate with 2X SYBR Green Premix Pro Taq HS qPCR Tracking Kit (ROX2 plus, AG11735, Accurate Biotechnology (Hunan) Co., Ltd), primers and template DNA.

Techniques: Expressing