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Sauer"],"technology_type":["Mass Spectrometry","Bottom-up proteomics","Data-independent acquisition","SWATH MS"],"disease":["Colon Cancer"],"software":[""],"submitter_keywords":["Tcipa","Proteomics","Colorectal cancer","Dia-ms","Metastasis","Platelets","Emt"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD062043"],"tissue":["Blood Platelet","Cell Culture"],"sample_protocol":["SW480 and SW620 cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured as previously described. Cells were detached using AccumaxTM (PAN-Biotech GmbH, Germany), washed with PBS, and resuspended in Tyrode’s buffer (140 mM NaCl, 3 mM KCl, 16.62 mM NaHCO3, 1 mM MgCl2·6H2O, 10 mM HEPES, 5.5 mM D-(+)-glucose in MilliQ water, pH 7.4). The tumor cell suspension was adjusted to 5 × 10⁵ cells/mL. Stable isotope labeling by amino acids in cell culture (SILAC) was used to differentiate tumor cell proteins from platelet proteins in mass spectrometry analyses. SW480 and SW620 cells were cultured in media containing isotopically labeled arginine (^13C^15N-arg) and lysine (^13C^15N-lys), introducing mass shifts of 8 Da and 10 Da for lysine and arginine, respectively. SILAC RPMI medium (SILANTES, Munich, Germany) was supplemented with labeled amino acids (arginine 200 mg/L, lysine 41.75 mg/L), 10% dialyzed FBS, glutamine (40 mg/L), and 1% penicillin/streptomycin. Cells were cultured until a labeling incorporation rate of ≥95% was achieved.  To confirm incorporation efficiency, cells were centrifuged, washed with PBS, and lysed using the EasyPep Mini MS Sample Prep Kit (Thermo Fisher Scientific). Light transmission aggregometry (LTA) was performed with an APACT 4S Plus aggregometer (DiaSys Greiner GmbH). Platelet samples were pre-incubated at 37°C for five minutes with a final CaCl2 concentration of 1 mM. Aggregation was induced using either TRAP-6 (12.5 μM) or tumor cell suspensions (20 μL containing isotopically labeled SW480 or SW620 cells at 2,500 cells/μL). Controls included platelets incubated with CaCl2 alone or with TRAP-6 as positive controls. Samples were centrifuged for 15 seconds post-collection and lysed using the EasyPep Mini MS Sample Prep Kit before storage at -80°C.  Protein concentrations were measured using the EZQ Protein Quantitation Kit (Invitrogen). Proteins (25–100 μg per sample) were digested with trypsin/Lys-C protease mix provided in the EasyPep Mini MS Sample Prep Kit. Peptides were purified using columns, frozen at -80°C, and lyophilized for storage at -20°C until reconstitution. Samples were solubilized to a final concentration of 1 µg/mL in solvent A (1% formic acid in HPLC/MS ultrapure water) and analyzed via a Dionex Ultimate 3000 HPLC system coupled to a TripleTOF 5600+ mass spectrometer (AB Sciex). Peptides were first desalted on a trap column (Acclaim PepMap C18) and separated on an analytical column (Luna C18(2)) using a gradient of solution B (0.1% formic acid in acetonitrile).  Mass spectrometry analyses employed SWATH-MS acquisition with electrospray ionization (ESI). Parameters included ion spray voltage of 5,000 V, curtain gas at 30 psi, collision energy of 25.1 eV with a spread of ±5 eV, and precursor isolation windows calculated via the SWATH variable window calculator. MS1 scans covered a mass range of 350–1,250 m/z with a 50 ms accumulation time followed by MS2 scans across Q1 windows.  DDA-MS measurements assessed isotopic labeling efficiency and informed SWATH method development. Identical instrument parameters were used for MS1 scans over a range of 350–1,250 Da with a longer accumulation time of 250 ms followed by MS2 scans from 100–1,500 Da at high sensitivity mode."],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["SWATH®-MS raw data of the platelet and tumor cell samples were processed using the multiplex functionality DIA-NN V1.8.1 software (data-independent acquisition by neural networks) (21, 22). Independent analysis runs were performed to generate distinct protein lists for platelets and tumor cells. An in-silico spectral library was predicted from the human UniProtKB Swiss-Prot database FASTA file using DIA-NN options for FASTA digest and deep learning-based prediction of spectra, retention times, and ion mobilities. Analysis of platelets and tumor cell proteins from raw MS/MS spectra was conducted primarily based on the default settings: Trypsin/P was specified as protease with a maximum of one missed cleavage. The maximum number of variable modifications was set to zero. N-terminal methionine excision and C-carbamidomethylation were enabled, whereas methionine oxidation, N-terminal acetylation, phosphorylation, and GG adduction on lysins were disabled and therefore not considered as variable modifications. The default settings were kept for the peptide length range (7-30 amino acids) and the precursor charge range (1-4). The precursor m/z range was set to 300-1,800 Da whereas the fragment ion m/z range was defined as 350-1,250 Da. The MS1 and MS2 accuracies were fixed to 15 and 20 Da, respectively. The ‘Match between runs’ and ‘No shared spectra’ options were enabled. Protein inference was performed based on genes, using single-pass mode as neural network classifier, and ´Any LC´ (high accuracy) quantification strategy. The cross-run normalization was RT-dependent. For tumor cell protein identification, SILAC labeling was accounted for using additional command line options: ‘--fixed-mod Lys8,8.014199,K,label’; ‘--fixed-mod Arg10,10.008269,R,label’; ‘--lib-fixed-mod Lys8’; ‘--lib-fixed-mod Arg10’; ‘--original-mods’ and ‘--gen-spec-lib’. The DIA-NN output was further processed using the DIA-NN R package (21). Filtering criteria included precursory with library q-value and library protein group q-value  0.01. The dataset was filtered for proteotypic proteins, and the MaxLFQ algorithm was used for protein quantification (23). Protein identified with less than two proteotypic peptides were excluded. The remaining proteins were finally mapped to their corresponding gene names for downstream analysis."],"omics_type":["Proteomics"],"labhead":["Prof. Dr. Timo Gemoll"],"instrument_platform":[""],"labhead_affiliation":["University of Lübeck Section for Translational Surgical Oncology & Biobanking Department of Surgery Ratzeburger Allee 160, BMF 23558 Lübeck, Germany"],"submission_type":["PARTIAL"],"species":["Homo Sapiens (human)"],"submitter_mail":["thorben.sauer@student.uni-luebeck.de"],"publication":["41896928 Sauer T, Gruner C, Kern K, Rackisch A, Tischner L, Schulz K, Ostermann J, Cohrs L, Kohl M, Verschoor A, Gemoll T. Proteomic remodeling during tumor cell-induced platelet aggregation unveils metastatic drivers in colorectal cancer. Cancer Cell Int. 2026 10.1186/s12935-026-04282-7"],"submitter_affiliation":["Department of Surgery, Laboratory for Surgical Research, University Hospital Schleswig-Holstein, Campus Lübeck, Ratzeburger Allee 160, 23538 Lübeck Germany"],"submitter_country":["Germany"],"additional_accession":[]},"is_claimable":false,"name":"Proteomic Remodeling During Tumor Cell-Induced Platelet Aggregation Unveils Metastatic Drivers in Colorectal Cancer","description":"Colorectal cancer (CRC) is frequently associated with metastasis, resulting in high mortality rates. Platelets are known to play a crucial role in the metastatic cascade influencing tumor microenvironment remodeling, promoting cell transformation, facilitating metastatic niche formation, and shielding circulating tumor cells from immune surveillance. However, platelet proteomic alterations during tumor cell-induced platelet aggregation (TCIPA) remain largely unexplored. This study aims to characterize the proteomic profile of TCIPA in CRC using an in vitro model that recapitulates key aspects of CRC metastasis. TCIPA was assessed via light transmission aggregometry using an in vitro model incorporating paired primary and metastatic cell cultures. Stable Isotope Labeling with Amino Acids in Cell culture (SILAC) allowed the discrimination of healthy platelet and tumor cell proteomes prior to and following TCIPA. Data-independent acquisition mass spectrometry was employed to analyze intra- and extracellular tumor and platelet proteomes. Comparative proteomic profiling was performed using a range of bioinformatic analyses, including clustering, differential expression, and Gene Set Enrichment Analyses (GSEA). Comparison of the baseline proteome profiles of SW480 and SW620 cell lines identified 263 significant differentially expressed proteins. The GSEA demonstrated enrichment of the ‘epithelial-mesenchymal transition’ gene set in SW480 cells. While SW480 exhibited rapid TCIPA, SW620 did not consistently interact with healthy platelets. Following TCIPA, 34 tumor proteins showed differential expression compared to their naïve status. Notably, 17 of these proteins were significantly associated with CRC progression, particularly in the promotion of EMT, metastasis, tumor cell survival, proliferation, and metabolic reprogramming. This study successfully characterized the proteomic profiles of platelets, platelet secretomes, and colorectal tumor cells following TCIPA-induced activation. The findings highlight the significant role of several tumor proteins and their metabolic effects in colorectal cancer progression.","dates":{"publication":"2026-04-02","submission":"2025-03-19"},"accession":"PXD062043","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:1126","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:247480","NEWT:1211601","NEWT:1323429","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","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:314344","NEWT:108061","NEWT:60711","NEWT:2020713","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:1220585","NEWT:3349","NEWT:160621","NEWT:2502409","NEWT:212790","NEWT:110368","NEWT:55350","NEWT:1916073","NEWT:33103","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:614666","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:1211620","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:709964","NEWT:242605","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:156974","NEWT:3714","NCBITaxon:37162","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: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: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: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:1392498","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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