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Gels were sliced in small pieces and destaining in 50% ethanol, 50% ammonium bicarbonate (ABC) Disulfide bonds were reduced with 10 mM dithiothreitol (DTT)and incubation at 56°C for 1 hour. Next, gel was treated with 55 mM iodoacetamide (IAA) incubated for 1 hour at room temperature in the dark for alkylation of cysteine residues. Then, gel pieces were washed with ethanol followed by ABC, and this step was repeated. After this step, 12.5 ng/µl trypsin in ABC was added to cover the gel inside the tube, with overnight incubation at 37°C. In the following day, peptides were extracted from the gel pieces by twice incubations in 3% TFA, 30% acetonitrile and twice incubations in 100% acetonitrile. The sample volume was reduced in vacuum centrifuge and the generated peptides were purified by reverse-phase chromatography using homemade C18 stage-tip. The stage-tips were pre-conditioned with 100% methanol and 0.1% formic acid. Samples were applied, followed by washing with 0.1% formic acid, and adsorbed peptides were eluted with 40% acetonitrile solution in 0.1% formic acid. The eluate was dried in vacuum centrifuge, resuspended in 0.1% formic acid and injected onto LC-MS/MS system. The samples were analyzed at Mass Spectrometry Facility (RPT02H), Technological Platforms Network FIOCRUZ, Oswaldo Cruz Foundation (FIOCRUZ) using an Orbitrap Fusion Lumos mass spectrometer (Thermo Scientific) coupled to an Ultimate 3000 RSLCnano (Thermo Scientific). Peptide separation was achieved using a 90-minute gradient at a flow rate of 250 nL/min. The elution gradient consisted of 5% to 40% solvent B (acetonitrile with 0.1% formic acid) over 90 minutes. The chromatographic system employed a 15 cm analytical column with 3 µm C18 particles (ReproSil-Pur 120 C18-AQ, Dr. Maisch) packed into a 75 µm (ID) × 360 µm (OD) FSC emitter. The mass spectrometer operated in Data Dependent Acquisition (DDA) mode with 2 s maximum per duty cycle, selecting the most intense precursor ions for Higher-Energy Collisional Dissociation (HCD) fragmentation and analysis in the orbitrap analyzer (MS/MS). Acquisition parameters included: nano-electrospray voltage set to 2.3 kV, , injection times of 50 ms and resolution of 120,000 for MS1 (300-1500 m/z) and 22 ms and 15,000 for MS2. The dynamic exclusion time was set to 60 seconds.The internal calibration option was enabled for MS1 scans."],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["Label-free quantification (LFQ) of protein abundance was obtained by processing the data on MaxQuant platform version 2.2.0.0 using the Canis lupus familiaris database (uniprot_proteome_UP000805418). Variable modifications included in the search were Oxidation (M); Acetyl (Protein N-term) Identified proteins with a false discovery rate (FDR) ≥ 1) % were filtered out."],"omics_type":["Proteomics"],"labhead":["Karine Araújo Damasceno"],"instrument_platform":[""],"submission_type":["PARTIAL"],"labhead_affiliation":["Laboratory for Global Health Research and Neglected Diseases, Gonçalo Moniz Institute, Oswaldo Cruz Foundation, BA, Brazil"],"species":["Canis Familiaris (dog) (canis Lupus Familiaris)"],"publication":["41952687 de Almeida BS, Rocha GV, Nunes S, Zanette DL, Batista M, Estrela-Lima A, Regis-Silva CG, Damasceno KA. Comparative omics reveals conserved extracellular matrix signatures during mammary tumor progression. Front Oncol. 2026 16:1788746 10.3389/fonc.2026.1788746"],"submitter_mail":["batista.mic@gmail.com"],"submitter_affiliation":["Fiocruz"],"submitter_country":["Brazil"],"pubmed_abstract":["The extracellular matrix (ECM) plays a critical role in tumor progression by modulating cell adhesion, migration, and signaling; however, its contribution to metastatic progression in spontaneous mammary tumors remains poorly understood. Mammary tumors are among the most common neoplasms in female dogs and share histopathological and molecular similarities with human breast cancer, supporting their use as a comparative oncology model. To investigate ECM remodeling during tumor progression, we analyzed normal, non-metastatic, and metastatic canine mammary tissues using histological approaches and label-free quantitative proteomics. Publicly available human breast cancer transcriptomic datasets were interrogated for <i>in silico</i> validation of conserved molecular signatures. Proteomic profiling identified 12 differentially expressed ECM-related proteins: eight were upregulated (COL12A1, COL4A1, COL4A2, SERPINH1, SERPINF1, HTRA1, TNC, PCOLCE) and four were downregulated (MMRN1, ABI3BP, DPT, OGN). The downregulated proteins were further validated in human breast cancer transcriptomes. Collectively, these findings indicate active ECM remodeling during tumor progression, characterized by increased expression of proteins associated with matrix stiffness and invasiveness. This study highlights evolutionarily conserved mechanisms of ECM dysregulation in breast cancer and identifies potential matrix targets for translational research and biomarker development."],"pubmed_title":["Comparative omics reveals conserved extracellular matrix signatures during mammary tumor progression."],"pubmed_authors":["de Almeida Bruno Sousa BS, Rocha Gisele Vieira GV, Nunes Simone S, Zanette Dalila Luciola DL, Batista Michel M, Estrela-Lima Alessandra A, Regis-Silva Carlos Gustavo CG, Damasceno Karine Araújo KA"],"additional_accession":[]},"is_claimable":false,"name":"Proteomic Analysis of Canine Extracellular Matrix in Mammary Tumor Progression","description":"Canine mammary gland tumors are common and bear striking similarities to human breast cancer, making them a valuable model for comparative oncology. This study examined changes in the extracellular matrix (ECM) associated with tumor progression in spontaneous canine mammary carcinomas, aiming to identify biomarkers linked to tumor aggressiveness. Normal, non-metastatic, and metastatic mammary tissues were analyzed by histological and proteomic approaches, and human breast cancer transcriptomic data were explored for in silico validation. Histological analysis with special stains revealed a reduction in type III collagen in metastatic tumors. The proteomic profile identified 12 MEC-related proteins with differential expressions in canine samples, of which eight were upregulated (COL12A1, COL4A1, COL4A2, SERPINH1, SERPINF1, HTRA1, TNC, PCOLCE) and four (MMRN1, ABI3BP, DPT, OGN) were downregulated and were confirmed to be differentially expressed in the human transcriptome as well. The results highlight the active remodeling of the ECM during tumor progression, marked by increased expression of proteins associated with stiffness and invasiveness and decreased expression of potential suppressors. These findings strengthen the understanding of ECM dynamics in breast cancer and suggest promising molecular targets for future investigations.","dates":{"publication":"2026-04-13","submission":"2026-02-09"},"accession":"PXD074203","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:108018","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","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: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: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:1916073","NEWT:1310161","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: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:3715","NEWT:2065263","NEWT:199225","NEWT:177437","NEWT:717966","NEWT:9014","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:29022","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:52299","NEWT:1883426","NEWT:8165","NEWT:96794","NEWT:10401","NEWT:10886","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: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: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:1116234","NEWT:415978","NEWT:235455","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:9054","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","NEWT:81231006","NEWT:156981","NEWT:30455","NEWT:28141","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:3728","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:219799","NEWT:219798","NEWT:548","NEWT:150475","NEWT:267872","NEWT:381666"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