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Stephane"],"technology_type":["Mass Spectrometry","All-ion fragmentation"],"disease":["Pancreatic Ductal Adenocarcinoma"],"software":[""],"submitter_keywords":["Monocytes","Immunosuppression","Lc-msms","Extracellular vesicles","Tumor-associated macrophages"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD039439"],"tissue":["Monocyte","Pancreatic Ductal Carcinoma Cell","Blood"],"sample_protocol":["Healthy donor blood, in concentrated buffy coats, was obtained from the French Blood Establishment (Marseille, France). Peripheral blood mononuclear cells (PBMC) were isolated using the Ficoll-Paque density gradient method. Briefly, buffy coats were mixed with RPMI 1640 medium (Gibco) supplemented with 10% FBS and 1% antibiotic-antimycotic and centrifuged at 400 x g for 40 minutes with Ficoll-Paque (Cytvia) to obtain a ring of PBMCs. This ring was isolated, incubated for 10 minutes in 1x red blood cell lysis buffer (eBioscience) and washed in 1X PBS 4 times before cell counting. CD14+ monocytes were isolated from PBMCs using magnetic labeling and sorting with human CD14 microbeads (Miltenyi Biotec) by positive selection following the manufacturer’s protocol and the autoMacs pro separator (Miltenyi Biotec). Monocytes were plated at a density of 500,000 cells per well in 24-well plates in RPMI medium containing 10% FBS depleted of EVs. Monocytes were treated with PBS (C: Control vehicle), PANC-1 (PE), MiaPaCa-2 (M) or healthy pancreatic ductal epithelial (HPDE)(H) EVs for two consecutive days. Two days following the last EVs treatment, plastic- and EVs-differentiated macrophages were detached. Cell pellets were washed twice with PBS and stored at -80°C until proteomic analysis. Blood from 3 donors (D1, D2, D3) were used for this experiment, each treated in triplicate with PANC-1 (PE), MiaPaCa-2 (M), healthy pancreatic ductal epithelial (HPDE)(H) EVs or PBS (C: Control vehicle).  For relative proteomic analysis, proteins from cell pellets were solubilized in 15µl of LDS sample buffer (2x concentrated), (Invitrogen, Life Technologies) before loading on NuPAGE™ 4–12% Bis–tris acrylamide gels according to the manufacturer’s instructions (Invitrogen, Life Technologies). Running of samples was stopped as soon as proteins stacked as a single band and following imperial blue staining (Life Technologies), the upper part of the gel containing the proteins was cut and processed for classical in gel digestion (washes, thiols reduction with 10 mM DTT and cystein alkylation with 55 mM iodoacetamide). Each band was further digested as previously described with trypsin and analyzed by liquid chromatography (LC)-tandem MS (MS/MS) using an Orbitrap Fusion Lumos Tribrid Mass Spectrometer (ThermoFisher Scientific, San Jose, CA) online with a nanoRSLC Ultimate 3000 chromatography system (ThermoFisher Scientific, Sunnyvale, CA). First, peptides were concentrated and purified on a pre-column PepMap100 C18, 2cm × 100µm I.D, 100Å pore size, 5µm particle size in solvent A (0.1% formic acid in 2% acetonitrile). In the second step, peptides were separated on a reverse phase LC EASY-Spray C18 column PepMap RSLC C18, 50cm × 75µm I.D, 100Å pore size, 2µm particle size (ThermoFisher) at 300nL/min flow rate and 40°C. After column equilibration using 4% of solvent B (20% water - 80% acetonitrile - 0.1% formic acid), peptides were eluted from the analytical column by a three-step linear gradient (2-20% solvent B for 90min, 20-45% for 20min and a last elution 45-90% for 10mn). For peptide ionization in the EASY-Spray nanosource in front of mass spectrometer, spray voltage was set at 2.2 kV and the capillary temperature at 275 °C. The Orbitrap Lumos was used in data-independent mode with the following parameters. First, MS spectra were acquired in the Orbitrap in the range of m/z 375-1500 at a FWHM resolution of 120,000 measured at 400 m/z. AGC target was set at standard parameters with an automatic Maximum Injection Time. MS2 spectra were acquired in the Orbitrap with a resolution of 30,000, in the mass range of 200-1800 m/z after isolation of parent ion in the quadrupole and fragmentation in the HCD cell under collision Energy of 30%. DIA parent ion range was from 400 to 1000 m/z divided into 40 windows 16 Da wide and from 1000 to 1500 m/z divided into 10 windows 50 Da wide."],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["Relative intensity-based label-free quantification (LFQ) was processed using the DIA-NN 1.8 algorithm. Raw files were searched against the Human database extracted from UniProt on the 2th of February 2022 and containing 20395 entries (reviewed) with the addition of a protein contaminant bank (J Proteome Res 2022;21(9):2104-2113. Protein Contaminants Matter: Building Universal Protein Contaminant Libraries for DDA and DIA Proteomics; Ashley M Frankenfield, Jiawei Ni, Mustafa Ahmed, Ling Hao). The following parameters were used for searches: (i) trypsin allowing cleavage before proline; (ii) one missed cleavages were allowed; (iii) cysteine carbamidomethylation (+57.02146) as a fixed modification and methionine oxidation (+15.99491) and N-terminal acetylation (+42.0106) as variable modifications; (iv) a maximum of 1 variable modification per peptide allowed; and (v) minimum peptide length was 7 amino acids and a maximum of 30 amino acids. The match between runs option was enabled to transfer identifications across different LC-MS/MS replicates based on their masses and retention time. The precursor false discovery was set to 1%. DIA-NN parameters were set on Single-pass mode for Neural Network classifier, Robust LC High precision for quantification strategy and RT-dependent mode for Cross-run normalization. Library was generated using Smart profiling set up. Main output file from DIA-NN was further filtered at 1% FDR and LFQ intensity was calculated using the iq package at 1% q-value (Bioinformatics 2020;36(8):2611-2613. iq: an R package to estimate relative protein abundances from ion quantification in DIA-MS-based proteomics; Thang V Pham, Alex A Henneman, Connie R Jimenez).  The statistical analysis was done with Perseus program (version 1.6.15.0) from the MaxQuant environment (www.maxquant.org). Quantifiable proteins were defined as those detected in above 70% of samples in one condition or more. Missing values were replaced using data imputation by randomly selecting from a normal distribution centered on the lower edge of the intensity values that simulates signals of low abundant proteins using default parameters (a downshift of 1.8 standard deviation and a width of 0.3 of the original distribution). To determine whether a given detected protein was specifically differential, a two-sample t-test was done using permutation-based FDR-controlled at 0.05 and employing 250 permutations. The p value was adjusted using a scaling factor s0 with a value of 1. Analysis was done on biological triplicates, each injected twice on mass spectrometers. One sample named D3MP2 was deliberately eliminated for final quantitative analysis as it appeared clearly as an outsider regarding PCA, multi scatterplots and imputation on distribution of intensity."],"omics_type":["Proteomics"],"labhead":["Richard Tomasini, PhD"],"instrument_platform":[""],"labhead_affiliation":["INSERM U1068/UMR7258 CNRS - Cancer Research Center of Marseille \"Pancreatic Cancer\" Team Group leader of the \"Microenvironment and inter-cellular dialogue\" Parc Scientifique de Luminy, 163 avenue de Luminy, case 915, 13288 Marseille Cedex 9 France"],"submission_type":["PARTIAL"],"species":["Homo Sapiens (human)"],"publication":["Not available"],"submitter_mail":["stephane.audebert@inserm.fr"],"submitter_affiliation":["Marseille Proteomic, Centre de Recherche en Cancérologie de Marseille, Inserm UMR1068, CNRS UMR7258, Aix Marseille Université U105, Institut Paoli Calmettes, 27 Boulevard Leï Roure CS30059 13273 Marseille Cedex 09 France"],"submitter_country":["France"],"additional_accession":[]},"is_claimable":false,"name":"Proteomic profile of healthy human monocytes differentiated to macrophages with non-tumor or tumor cell-derived extracellular vesicles","description":"To investigate the impact of tumor cell-derived extracellular vesicles on monocyte-macrophage differentiation, healthy monocytes were treated with EVs derived from two different pancreatic cancer cell lines and a non-tumoral pancreatic ductal epithelial cell line. Following differentiation to macrophages, proteomic analyses by mass spectrometry were performed on cells.","dates":{"publication":"2026-06-11","submission":"2023-01-13"},"accession":"PXD039439","cross_references":{"TAXONOMY":["NEWT:108018","NEWT:377960","NEWT:1131","NEWT:190486","NEWT:2042546","NEWT:148942","NEWT:295546","NEWT:258594","NEWT:1126","NEWT:1129","NEWT:89187","NEWT:309807","NEWT:89184","NEWT:309800","NEWT:1323429","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:44270","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:10034","NEWT:661410","NEWT:488003","NEWT:224326","NEWT:376619","NEWT:3765","NEWT:2433","NEWT:3767","NEWT:433727","NEWT:135622","NEWT:672127","NEWT:656061","NEWT:37965","NEWT:656064","NEWT:3760","NEWT:36630","NEWT:381198","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","NEWT:44250","NEWT:317447","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:12637","NEWT:2164133","NEWT:1180","NEWT:2767846","NEWT:1343074","NEWT:314344","NEWT:108061","NEWT:224308","NEWT:49928","NEWT:160621","NEWT:110368","NEWT:1916073","NEWT:1310161","NEWT:1230383","NEWT:77133","NEWT:1310165","NEWT:114727","NEWT:5516","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:1410384","NEWT:1310173","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:419481","NEWT:5507","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:1143","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:709964","NEWT:3712","NEWT:3711","NEWT:3714","NEWT:65121","NEWT:3715","NEWT:2065263","NEWT:1549675","NEWT:177437","NEWT:9014","NEWT:10891","NEWT:118221","NEWT:34869","NEWT:226126","NEWT:1616117","NEWT:34865","NEWT:28174","NEWT:550537","NEWT:284812","NEWT:3701","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:1246791","NEWT:34875","NEWT:9005","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:1883426","NEWT:657321","NEWT:1423687","NEWT:10886","NEWT:3708","NEWT:536231","NEWT:219813","NEWT:1301262","NEWT:2305479","NEWT:215451","NEWT:460519","NEWT:272843","NEWT:272844","NEWT:1348799","NEWT:55784","NEWT:485","NEWT:2499525","NEWT:467144","NEWT:142786","NEWT:2853422","NEWT:487","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:46835","NEWT:216778","NEWT:376686","NEWT:237610","EFO:0001352","NEWT:219809","NEWT:219808","NEWT:119562","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:141469","NEWT:272831","NEWT:987059","NEWT:491","NEWT:460523","NEWT:2423","NEWT:85694","NEWT:85692","NEWT:911045","NEWT:3750","NEWT:291146","NEWT:233271","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:278161","NEWT:415978","NEWT:410289","NEWT:191701","NEWT:9054","NEWT:472","NEWT:29459","NEWT:1380096","NEWT:28128","NEWT:470","NEWT:41122","NEWT:1328300","NEWT:748693","NCBITaxon:50557","NEWT:3735","NEWT:29491","NEWT:911022","NEWT:446","NEWT:153481","NEWT:714485","NEWT:445","NEWT:2003758","NEWT:29486","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:215402","NEWT:3726","NEWT:30455","NEWT:28141","NEWT:9030","NEWT:9031","NEWT:3728","NEWT:108931","NEWT:219799","NEWT:1325292","NEWT:219798","NEWT:548","NEWT:150475","NEWT:256737","NEWT:51350","NEWT:8694","NEWT:8692","NEWT:542","NEWT:394503","NEWT:13076","NEWT:219785","NEWT:451515","NEWT:8673","NEWT:550","NEWT:10804","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:1225786","NEWT:771870","NEWT:8665","NEWT:51338","NEWT:9995","NEWT:28112","NEWT:49012","NEWT:28110","NEWT:10832","NEWT:311339","NEWT:507601","NEWT:520","NEWT:64657","NEWT:186441","NEWT:2585209","NEWT:643680","NEWT:214092","NEWT:40697","NEWT:13095","NEWT:162425","NEWT:242124","NEWT:9986","NEWT:8654","NEWT:536","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:80833","NEWT:479096","NEWT:28108","NEWT:28104","NEWT:1114957","NEWT:568703","NEWT:568708","NEWT:52641","NEWT:353152","NEWT:116597","NEWT:353153","NEWT:116598","NEWT:227377","NEWT:1774284","NEWT:27689","NEWT:207769","NEWT:40674","NEWT:51329","NCBITaxon:1763","NEWT:443906","NEWT:13037","NEWT:519","NEWT:941638","NEWT:7394","NEWT:6063","NEWT:404330","NEWT:1328388","NEWT:13064","NEWT:300641","NEWT:39655","NEWT:38323","NEWT:206411","NEWT:80869","NEWT:214053","NEWT:80863","NEWT:665890","NEWT:52639","NEWT:52638","NEWT:8697","NEWT:861109","NEWT:8695","NEWT:1196866","NEWT:6039","NEWT:1045775","NEWT:1123869","NCBITaxon:1335626","NEWT:7370","NEWT:280406","NEWT:83906","NEWT:99488","NEWT:99487","NEWT:1134506","NEWT:255470","NEWT:762903","NEWT:27697","NEWT:86185","NEWT:9935","NEWT:266940","NEWT:287889","NEWT:9938","NEWT:1028307","NEWT:52603","NEWT:92867","NEWT:92866","NEWT:1344414","NEWT:202950","NEWT:741158","NEWT:1293091","NEWT:202956","NEWT:483179","NEWT:315750","NEWT:99490","NEWT:632957","NEWT:11294","NEWT:9925","NCBITaxon:9606","NEWT:1436183","NEWT:1479235","NEWT:4232","NEWT:9922","NEWT:416348","NEWT:105812","NEWT:11298","NEWT:4233","NEWT:4236","NEWT:398031","NEWT:196627","NEWT:523840","NEWT:487521","NEWT:1076878","NEWT:200308","NEWT:200302","NEWT:9913","NEWT:9915","NEWT:105841","NEWT:1170227","NEWT:2666255","NEWT:999810","NEWT:28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