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NP0321PK2). Five adjacent bands were excised and subjected to trypsin digestion following the protocol by Shevchenko et al., with minor adjustments, as follows: protein reduction in 10 mM DTT for 1 hour at 56º C; protein alkylation with 55 mM iodoacetamide for 45 minutes at room temperature in the dark and digestion with 12.5 ng/µl trypsin overnight at 37ºC. Tryptic peptides were extracted from the gel with 3% trifluoroacetic acid (TFA) and 30% acetonitrile (ACN). The extracted peptides were de-salted and concentrated onto STAGE Tips microcolumn. ACN was evaporated using a speed vac concentrator (Eppendorf) and the volume of the eluates were adjusted to 5 µl with 0.1% FA, to be then analysed by online nano-flow liquid chromatography tandem mass spectrometry (nLC-MS/MS) using an EASY-nLC 1500 (Thermo Fisher Scientific) connected to a Q-Exactive HF instrument (Thermo Fisher Scientific) through a nano-electrospray ion source. The nano-LC system was operated in one column set-up with an EasySpray PEPMAP RSLC C18 column (Thermo Fisher Scientific) kept at 45°C constant. Solvent A was 0.1% formic acid (FA) and solvent B was 0.1% FA in 80% ACN. Samples were injected in aqueous 1% (Trifluoroacetic acid, TFA) at a constant pressure of 980 Bar. Peptides were separated with a gradient of 3–35% solvent B over 49 min followed by a gradient of 30–60% for 5 min and 60–95% over 5 min at a flow rate of 300 nL/min. The Q-Exactive was operated in the data-dependent acquisition (DDA) to automatically switch between full scan MS and MSMS acquisition. MS spectra (from m/z 375-1650) were analyzed in the Orbitrap detector with resolution R=70 000 at m/z 400. The 12 most intense peptide ions with charge states ≥2 were sequentially isolated to a target value of 36 and fragmented with a normalized collision energy setting of 28% in to the HCD cell. The maximum allowed ion accumulation times were 20 ms for full scans and 80 ms for MSMS."],"repository":["Pride"],"modification":[""],"quantification_method":["Not available"],"data_protocol":["Acquired raw data obtained by mass spectrometry analysis were analyzed using the MaxQuant (MQ) version 1.6.10.43, and peptide lists were searched against the human Uniprot FASTA database (74470 Entries) with the Andromeda search engine. False discovery rate (FDR) for both protein and peptide identifications was set to a maximum of 1% with enzyme specificity set to Trypsin/P. A maximum of 2 missed cleavages was allowed, and the minimum peptide length was fixed at 7 amino acids and carbamidomethylation of Cysteine was specified as a fixed modification. Peptides were identified with an initial precursor mass deviation of 7 ppm and a fragment mass deviation of 20 ppm. For label-free protein quantitation, the LFQ intensity calculation was enabled, as well as the match between runs (MBRs) feature. The “protein groups” output file from MaxQuant was analysed using Perseus software. Briefly, “contaminants” and “reverse” hits were manually remuved from the data set. Missing values were then imputed by replacing them with random numbers drawn from a normal distribution, assuming they originated from the low-intensity spectrum of the distribution (downshift = 1.8, width = 0.3). Statistically significant enriched proteins were identified using differential analysis with a threshold setting of S0 = 1 and FDR = 0.05, comparing MOMO versus Control experiments"],"omics_type":["Proteomics"],"labhead":["Eleonora Leucci"],"instrument_platform":[""],"submission_type":["PARTIAL"],"labhead_affiliation":["Laboratory for RNA Cancer Biology, Department of Oncology, KU Leuven, Leuven, Belgium; Department of Oncology and Hematology-Oncology, University of Milan, Milan, 20122 Italy"],"species":["Homo Sapiens (human)"],"publication":["Not available"],"submitter_mail":["alessandro.cuomo@ieo.it"],"submitter_affiliation":["IEO"],"submitter_country":["Italy"],"additional_accession":[]},"is_claimable":false,"name":"Generation of drug-tolerant cells by lncRNAs downstream of osmotic stress","description":"Environmental cues within the tumor microenvironment (TME) affect tumor heterogeneity and plasticity, ultimately influencing therapy responses. Recent studies have shown that cancer stem cell niches are located near blood vessels and thus may be exposed to abrupt changes in the osmotic conditions. Here, we investigated how osmotic stress induced by the TME contributes to the generation of drug-tolerant persister cells in melanoma. Our research provide evidence that osmotic stress promotes cancer cell adaptation and reprogramming toward an NCSC-like state, with long non-coding RNAs (lncRNAs) emerging as likely (and druggable) mediators of this mechanical stress, converting osmotic gradients into specific molecular signals. Critically, we have identified MOMO (Melanoma Osmosis MediatOr) as a novel transcript deregulated upon osmotic stress and associated with poor prognosis in melanoma. Our findings suggest that MOMO plays a crucial role in osmotic stress-induced adaptive reprogramming in melanoma by acting as a key player in the DNA Damage Response, thereby altering tumor composition and impacting therapy outcomes.","dates":{"publication":"2026-05-31","submission":"2025-01-30"},"accession":"PXD060369","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","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:1073882","NEWT:6819","NEWT:475174","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","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:656064","NEWT:1582","NEWT:3760","NEWT:1584","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:7719","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:3349","NEWT:160621","NEWT:212790","NEWT:110368","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","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:2579791","NEWT:44685","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","NEWT:410661","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:3714","NCBITaxon:37162","NEWT:270643","NEWT:2065263","NEWT:177437","NEWT:717966","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: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:8165","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2499525","NEWT:2853422","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:119562","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:108458","NEWT:5936","NEWT:321967","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:415978","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:28128","NEWT:470","NEWT:41122","NEWT:5911","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:11723","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:3726","NEWT:906914","NEWT:81231006","NEWT:28141","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:3728","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:548","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:2126348","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:542","NEWT:2903","NEWT:185579","NEWT:872227","NEWT:13076","NEWT:2587410","NEWT:207340","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:2126337","NEWT:550","NEWT:33941","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:771870","NEWT:51338","NEWT:9995","NEWT:575412","NEWT:28112","NEWT:49012","NEWT:6493","NEWT:10832","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","NEWT:64657","NEWT:186441","NEWT:643680","NEWT:214092","NCBITaxon:6157","NEWT:13095","NEWT:162425","NEWT:104105","NEWT:216257","NEWT:9986","NEWT:8654","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:5147","NEWT:2810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