{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Xlsx":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD074117/SampleAnnotation.xlsx"],"Tabular":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD074117/report.pr_matrix.tsv","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD074117/report-lib.tsv","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD074117/report.pg_matrix.tsv","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD074117/report-lib.tsv.speclib"],"Txt":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD074117/checksum.txt"],"Wiff":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD074117/Batch2.IBD.DIA.Wiff.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD074117/Batch1.IBD.DIA.Wiff.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD074117/Batch3.IBD.DIA.Wiff.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD074117/Batch4.IBD.DIA.Wiff.zip"],"Other":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD074117/FASTA.2022-10-06-reviewed-isoforms-contam-UP000005640-spikein.fas"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"labhead_mail":["Jean-Francois.Beaulieu@USherbrooke.ca"],"submitter":["Elmira Shajari"],"technology_type":["Mass Spectrometry","Data-independent acquisition","SWATH MS"],"disease":["Inflammatory Bowel Disease 7"],"software":["Not available"],"submitter_keywords":["Inflammatory bowel disease","Biomarker discovery","Human","Stool","Proteomics","Machine learning","Clinical mass spectrometry","Nested cross validation","Ibd activity","Peptidomics"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD074117"],"sample_protocol":["2.1 Stool sample collection Between January 2021 and August 2023, fecal specimens were sourced through the routine fecal calprotectin program of the Clinical Hematology Laboratory at CIUSSS de l’Estrie–CHUS. Participants collected samples at home and delivered them to the hospital within 24 h (up to ~2 h at room temperature, then refrigerated at 4 °C for ≤24 h). Approximately 50 mg of each specimen was allocated for ELISA-based calprotectin testing; the remaining material was aliquoted and stored at −80 °C for proteomic analyses, following patient consent.  Sample preparation and DIA acquisition followed our previously published protocol, with minor adaptations. Briefly, ~100 mg of frozen stool was suspended in lysis buffer (25 mM Tris, 1% SDS, pH 7.5) and centrifuged; protein content in the aqueous phase was quantified by BCA assay. Proteins were reduced with 10 mM DTT, alkylated to 15 mM iodoacetamide, and the reaction was quenched with 10 mM DTT. Precipitation was performed using chilled acetone and methanol. The protein pellet was digested with Trypsin/Lys-C, and resulting peptides were cleaned on Strata-X SPE cartridges, dried at 37 °C for ~45 min, and stored at −80 °C. Prior to LC–MS/MS, peptides were reconstituted in 20 µL of mobile phase (0.2% formic acid, 3% DMSO in water)."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["2.8 Statistical analysis for discovering potential biomarkers The key steps of the analytical workflow are summarized in Figure 1. All analyses were performed in R 4.2.2 (2022-10-31, UCRT) within RStudio. In total, 174 stool samples were analyzed (66 active IBD, 108 remission), including 34 samples with fecal calprotectin (FC) values in the gray zone (100–250 µg/g). Data preprocessing followed our previously described protocols (8): Raw SWATH-DIA intensities at the protein and peptide levels were log2-transformed and processed as previously described (9). Briefly, samples and features (protein/peptide) with more than 70% missing values were removed, and the remaining data were quantile-normalized (preprocessCore (14)). To assess batch structure, we first used exploratory boxplots across the four LC–MS acquisition batches and principal component analysis (PCA), Batch effects were then corrected using ComBat (sva (15)), after which intensities were re-inspected by boxplots and PCA to confirm improved alignment across batches. For the proteome-level description, we first imputed remaining missing values in the batch corrected protein matrix using k-nearest neighbors (impute (16)). For each protein, we then calculated the log2 fold change between Active and Remission samples and performed a two-sample Welch’s t-test. P-values were adjusted for multiple testing using the Benjamini–Hochberg procedure, and proteins with adjusted p < 0.05 and |log2FC| > 0.6 were classified as significantly up- or down-regulated; all others were considered non-significant. These statistics were used to generate volcano plots of log2FC versus −log10 (adjusted p-value), with the most significant differentially expressed proteins annotated, and violin plots for selected proteins to illustrate group-wise abundance differences and the degree of overlap between Active and Remission samples."],"omics_type":["Proteomics"],"labhead":["Jean-Francois Beaulieu"],"instrument_platform":[""],"labhead_affiliation":["Laboratory of Intestinal Physiopathology, Department of Immunology and Cell Biology, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, QC, Canada"],"submission_type":["PARTIAL"],"species":["Homo Sapiens (human)"],"submitter_mail":["elmira.shajari@usherbrooke.ca"],"publication":["42052152 Shajari E, Gagné D, Malick M, Roy P, Noël JF, Gagnon H, Delisle M, Boisvert FM, Brunet MA, Beaulieu JF. Non-invasive assessment of inflammatory bowel disease activity using a DIA-derived stool peptidomic signature and machine learning. Front Mol Biosci. 2026 13:1768474 10.3389/fmolb.2026.1768474"],"submitter_affiliation":["PhD candidate"],"submitter_country":["Canada"],"pubmed_abstract":["<h4>Introduction</h4>Monitoring disease activity in inflammatory bowel disease (IBD) is essential for guiding therapy and preventing irreversible tissue damage. Colonoscopy, although the gold standard, is invasive and unsuitable for frequent monitoring, while fecal calprotectin lacks accuracy within its diagnostic gray zone (fecal calprotectin 100-250 μg/g). Stool proteomics offers a non-invasive alternative by directly capturing molecular signatures of intestinal inflammation. We conducted a proof-of-concept study to determine whether stool-derived peptides can accurately classify IBD activity (Active vs. Remission) using a fully unbiased and reproducible nested cross-validation machine-learning framework.<h4>Methods</h4>A total of 174 stool samples from IBD patients were collected and profiled using SWATH-DIA mass spectrometry. Feature selection was performed within the training loops only (Boruta, LASSO, RFE) across repeated subsampling, retaining peptides consistently identified in ≥70% of runs. Stable features were used to train four classifiers (GLMNet, SVM-Radial, SVM-Linear, Naïve Bayes) under inner 5-fold tuning. Outer test folds provided fully unseen evaluation, and model performance was additionally assessed exclusively on gray zone samples extracted from the outer test splits to quantify diagnostic resolution in this clinically challenging subgroup.<h4>Results</h4>Nested cross-validation identified a consensus panel of nine stool-derived peptides from five proteins. Across candidate classifiers, performance was broadly similar, with GLMNet consistently achieving the best trade-off between metrics. For GLMNet, outer-fold mean AUC was 0.93 and balanced accuracy 0.88, with specificity 0.94, sensitivity 0.82, and F1-score 0.85; close agreement between inner- and outer-fold metrics indicated minimal overfitting. Within the calprotectin gray zone subgroup (n = 34), GLMNet maintained good performance (balanced accuracy 0.78, F1 0.79, AUC 0.80), confirming that the peptide signature remains informative in this diagnostically challenging range.<h4>Conclusion</h4>A stool-based multi-peptide signature, evaluated with a rigorously nested, leakage-free machine-learning framework, can reliably classify IBD activity and retain discriminative power within the gray zone. This biologically interpretable five-protein panel provides a strong basis for targeted mass-spectrometry assay development and prospective validation as a non-invasive tool for personalized IBD monitoring."],"pubmed_title":["Non-invasive assessment of inflammatory bowel disease activity using a DIA-derived stool peptidomic signature and machine learning."],"pubmed_authors":["Shajari Elmira E, Gagné David D, Malick Mandy M, Roy Patricia P, Noël Jean-François JF, Gagnon Hugo H, Delisle Maxime M, Boisvert François-Michel FM, Brunet Marie A MA, Beaulieu Jean-François JF"],"additional_accession":[]},"is_claimable":false,"name":"Identification of a Stable Stool Peptidomic Signature for Monitoring IBD Activity","description":"Introduction Monitoring disease activity in inflammatory bowel disease (IBD) is essential for guiding therapy and preventing irreversible tissue damage. Colonoscopy, although the gold standard, is invasive and unsuitable for frequent monitoring, while fecal calprotectin lacks accuracy within its diagnostic gray zone (fecal calprotectin 100–250 µg/g). Stool proteomics offers a non-invasive alternative by directly capturing molecular signatures of intestinal inflammation. We conducted a proof-of-concept study to determine whether stool-derived peptides can accurately classify IBD activity (Active vs Remission) using a fully unbiased and reproducible nested cross-validation machine-learning framework. Methods A total of 174 stool samples from IBD patients were collected and profiled using SWATH-DIA mass spectrometry. Feature selection was performed within the training loops only (Boruta, LASSO, RFE) across repeated subsampling, retaining peptides consistently identified in ≥70 % of runs. Stable features were used to train four classifiers (GLMNet, SVM-Radial, SVM-Linear, Naïve Bayes) under inner 5-fold tuning. Outer test folds provided fully unseen evaluation, and model performance was additionally assessed exclusively on gray zone samples extracted from the outer test splits to quantify diagnostic resolution in this clinically challenging subgroup. Results Nested cross-validation identified a consensus panel of nine stool-derived peptides from five proteins. Across candidate classifiers, performance was broadly similar, with GLMNet consistently achieving the best trade-off between metrics. For GLMNet, outer-fold mean AUC was 0.93 and balanced accuracy 0.88, with specificity 0.94, sensitivity 0.82, and F1-score 0.85; close agreement between inner- and outer-fold metrics indicated minimal overfitting. Approximately half of the misclassified test patients were shared with the other models, suggesting intrinsically ambiguous cases rather than GLMNet-specific errors. Within the calprotectin gray zone subgroup (n = 34), GLMNet maintained good performance (accuracy 0.76, balanced accuracy 0.78, F1 0.79, AUC 0.80), confirming that the peptide signature remains informative in this diagnostically challenging range. SHAP and correlation analyses confirmed directionally consistent, largely non-redundant peptide contributions, and network analysis showed that the underlying proteins participate in related but distinct inflammatory pathways. Conclusions A stool-based multi-peptide signature, evaluated with a rigorously nested, leakage-free machine-learning framework, can reliably classify IBD activity and retain discriminative power within the gray zone. This biologically interpretable five-protein panel provides a strong basis for targeted mass-spectrometry assay development and prospective validation as a non-invasive tool for personalized IBD monitoring.","dates":{"publication":"2026-05-25","submission":"2026-02-06"},"accession":"PXD074117","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:108018","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:1126","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:247480","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:1367493","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:46429","NEWT:475174","NEWT:1599","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:2736291","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:294671","NEWT:376619","NEWT:1748418","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:135622","NEWT:4615","NEWT:672127","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656061","NEWT:37965","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:2086595","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:381198","NEWT:418106","NCBITaxon:12721","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:44250","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:3357","NEWT:7719","NEWT:29073","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:2767846","NEWT:108061","NEWT:60711","NEWT:2020713","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:3349","NEWT:160621","NEWT:2502409","NEWT:212790","NEWT:110368","NEWT:55350","NEWT:1916073","NEWT:1310161","NEWT:1230383","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:468822","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","NEWT:370038","NEWT:498211","NEWT:345076","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:200795","NEWT:342808","NEWT:2579791","NEWT:44685","NEWT:161934","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","NEWT:410661","NEWT:329854","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:2649329","NEWT:1143","NEWT:99802","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:709964","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:156974","NEWT:3714","NCBITaxon:37162","NEWT:270643","NEWT:3715","NEWT:2065263","NEWT:199225","NEWT:1549675","NEWT:177437","NEWT:717966","NEWT:9014","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:28174","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:29022","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:96795","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:9005","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:52299","NEWT:1883426","NEWT:8165","NEWT:96794","NEWT:10401","NEWT:10886","NEWT:686340","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:1955249","NEWT:215451","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:2714896","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:1608994","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:329032","NEWT:56636","NEWT:2499525","NEWT:55307","NEWT:1122981","NEWT:2853422","NEWT:1122984","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:2830","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:1502","NEWT:128017","NEWT:2836","NEWT:376686","NEWT:237610","NEWT:95486","EFO:0001352","NEWT:219809","NEWT:219808","NEWT:119562","NEWT:1883446","NEWT:31345","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:141469","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:1578","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:1922344","NEWT:108458","NEWT:5936","NEWT:85692","NEWT:321967","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:279010","NEWT:291146","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:278161","NEWT:1116234","NEWT:415978","NEWT:235455","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:9054","NEWT:1092586","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:28128","NEWT:470","NEWT:41122","NEWT:5911","NEWT:1328300","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:911022","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:717989","NEWT:33952","NEWT:1255232","NEWT:11723","NEWT:445","NEWT:153009","NEWT:1004879","NEWT:261756","NEWT:166314","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:1871049","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:3726","NEWT:906914","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