{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Txt":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/SDRF.txt"],"Mzml":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/Proteome-based_differences_between_drugs_within_fecal_cultures_of-one_CD_patient.mzML","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/Metaproteome-based_differences_between_6-mercaptopurine_exposure_and_control_comparing_five_CD_patients..mzML"],"Raw":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_B2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_A12.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_A8.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_A3.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_B6.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_B3.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_B10.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_B7.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_A9.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_A4.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_B4.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_B8.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_A1.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_A5.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_B11.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_B1.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_B5.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_A11.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_A2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_B9.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_B12.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_A7.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_A6.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/220329_HB_A10.raw"],"Mzid":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/Proteome-based_differences_between_drugs_within_fecal_cultures_of-one_CD_patient.mzid.gz","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD054714/Metaproteome-based_differences_between_6-mercaptopurine_exposure_and_control_comparing_five_CD_patients..mzid.gz"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"submitter":["Ronny Mohren"],"software":[""],"tissue":["Feces"],"sample_protocol":["Sample collection and fecal culture The participants collected complete defecations, using airtight boxes and one sachet AnaeroGen 1.5 L (Ref.: AN0025A, ThermoScientific). The sample was stored immediately at 4 °C until processing within 5 hours. The sample was transferred into an anaerobic chamber with 80 % Nitrogen, 10 % CO2, and 10 % H2 and homogenized in defined culture medium as reported by Li et al. with pH 7.813. The final incubations contained 2 % (w/v) fecal sample, 1 % dimethylsulfoxide, and were exposed to either 55 μg/ml budesonide (Ref.: B7777, Sigma-Aldrich), 42 μg/ml 6-mercaptopurine (Ref.: 852678, Sigma-Aldrich), 5 μg/ml (10 %) or 15 μg/ml (30 %) tofacitinib (Ref.: PZ0017, Pfizer®) or medium. We chose these concentrations based on the maximum oral intake distributed in 200 g colon content. Since 6-MP is dosed per kg body weight, we based our calculations on the approximate median body weight of CD patients, namely 70 kg19. All conditions were incubated in duplicate in a 2 ml 96-deep well plate covered with a PCR cover slip with ventilation holes and being placed in an IKA MS 3 basic plate shaker at 500 rpm for 24 hours at 37°C in the anaerobic chamber. Thereafter, samples were processed on ice and large debris removed by three times repeated centrifugation at 300 x g for 5 min. Then, the culture was pelleted and washed with PBS at 2272 x g for 1 hour at 4 °C as described by Li et al13. DNA was isolated as described elsewhere20, following the adapted protocol Q as described by the Human Microbiome Standards consortium21. Proteins were isolated by diluting washed pellets in 1 ml 0.01 % RapiGest SF (Ref.: 186001861, Waters) in 50 mM ammonium bicarbonate as done for DNA isolation. The supernatant was harvested after mechanical lysis and centrifugation at 16000 g. Proteomics analysis To explore microbial proteins and enzymatic pathways linked to drug exposure, we conducted proteomics analysis based on liquid chromatography followed by label-free quantitative mass spectrometry (LC-MS). Isolated proteins were reduced with 20 mM Dithiothreitol (DTT) for 45 minutes and alkylated with 40 mM Iodoacetamide for 45 minutes in the dark. Alkylation was terminated by 20 mM DTT. Subsequent protein digestion was performed using Endoproteinase LysC and Trypsin (Ref.: V5071, Promega), diluted 1:25 (enzyme to protein) for 2 hours at 37 °C and 750 rpm. For further overnight digestion, 50 mM ammoniumbicarbonate (ABC) was added to reach a final concentration of 1 M urea, and terminated by 1 % formic acid. LC-MS analysis was performed as described by Mezger et al.29, adapted slightly using a 110 minute gradient from 4 % to 32 % acetonitrile."],"repository":["Pride"],"labhead":["Ron Heeren / Ronny Mohren"],"instrument_platform":[""],"submission_type":["COMPLETE"],"labhead_affiliation":["Maastricht University, FHML, M4i"],"publication":["41826961 Becker HEF, Mohren R, Le NG, Derijks LJJ, Jonkers DMAE, Penders J. 6-mercaptopurine and tofacitinib alter microbial protein expression but not composition in fecal microbiota incubations from Crohn's disease patients. BMC Biol. 2026 24(1):101 10.1186/s12915-026-02569-9"],"submitter_affiliation":["Maastricht University"],"labhead_mail":["r.mohren@maastrichtuniversity.nl"],"technology_type":["Data-dependent acquisition","Mass Spectrometry","Bottom-up proteomics"],"disease":["Crohn's Disease"],"submitter_keywords":["Drugs","Lc-ms","Human","Proteomics","Crohn's disease","Microbiome"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD054714"],"quantification_method":[""],"modification":[""],"data_protocol":["For protein annotation, the human microbiome database was used as described by Li et al.30 and analyzed using Proteome Discoverer 2.5 (Thermofisher). For subsequent data analysis, False Discovery Rate (FDR) corrected (FDR < 1 %) and normalized values were used. KEGG orthology was applied for functional annotation using GhostKOALA with option “prokaryotes”31. Microbial taxonomic annotation based on identified peptides was conducted using Unipept 4.032."],"omics_type":["Proteomics"],"species":["Human Gut Metagenome","Gut Metagenome"],"submitter_mail":["r.mohren@maastrichtuniversity.nl"],"submitter_country":["Netherlands"],"doi":["10.6019/PXD054714"],"pubmed_abstract":["<h4>Background</h4>Crohn's disease (CD) is a chronic, relapsing-remitting gastrointestinal inflammatory condition with a multifactorial etiology. At present, drug therapy is the most important treatment option. However, a substantial number of CD patients experience side effects and/or nonresponse to medical drugs. In part, this might be attributed to the interaction of the intestinal microbiome with xenobiotics, such as medical drugs. The aim of this study was to explore the effect of the common CD drugs budesonide, 6-mercaptopurine (6-MP), as well as tofacitinib on the CD patient's microbiome in vitro.<h4>Results</h4>We performed 16S rRNA gene-based bacterial community profiling and metaproteomic analyses on anaerobic ex vivo incubations of CD patient-derived fecal microbiota (FM) that were exposed to CD drugs or control conditions. Both bacterial community profiling and metaproteomics revealed larger differences in 24-h FM incubations between the five donor-derived FM samples than between the various drug incubations. Incubation of the FM of one of the donors with 6-MP or tofacitinib resulted in a significant alteration in the metaproteome when compared to the control condition, whereas no effect could be observed upon incubation with budesonide. Considering only bacterial proteins detected in at least 80% of either the drug or control FM incubations, 33 proteins were consistently more abundant and 93 less abundant in all five donor-derived samples with 6-MP incubation, distinguishing 6-MP from control conditions. In contrast to metaproteomic analyses, bacterial community profiling only detected a significantly lower relative abundance of Colidextribacter in 15 µg/ml tofacitinib FM incubations. No alterations were detected in overall bacterial richness, diversity, or community structure in response to incubation with any of the drugs.<h4>Conclusions</h4>Tofacitinib and especially 6-MP significantly affect microbial function, but barely microbial composition in vitro. These drug-induced functional changes may subsequently influence host physiology and potentially inflammation in CD. Our findings emphasize the relevance to include functional microbial studies when investigating drug-microbiota interactions. Further research is needed to elucidate the impact of 6-MP-induced microbial alterations on intestinal physiology and inflammation in CD."],"pubmed_title":["6-mercaptopurine and tofacitinib alter microbial protein expression but not composition in fecal microbiota incubations from Crohn's disease patients."],"pubmed_authors":["Becker Heike E F HEF, Mohren Ronny R, Le N Giang NG, Derijks Luc J J LJJ, Jonkers Daisy M A E DMAE, Penders John J"],"additional_accession":[]},"is_claimable":false,"name":"Human microbiome LC-MS Proteomics in Crohn's disease patients","description":"Introduction. A substantial number of Crohn’s disease (CD) patients experience side-effects and/or non-response to medical drugs. In part, this might be attributed to the interaction of the intestinal microbiome with xenobiotics. The aim of this study was to explore the effect of common CD drugs on the patient’s microbiome in vitro. Methods The fecal microbiome of each of 5 CD patients was exposed to 42 μg/ml budesonide, 55 μg/ml 6-mercaptopurine (6-MP), 5 or 15 μg/ml tofacitinib, or DMSO-control in defined culture medium in an anaerobic chamber at 37°C for 24 hours. Subsequently, DNA and proteins were isolated and subjected to 16 rRNA gene amplicon sequencing and LC-MS proteomic analysis, 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