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Hamza Ewess"],"technology_type":["Affinity purification coupled with mass spectrometry proteomics","Mass Spectrometry"],"disease":["Influenza"],"software":[""],"submitter_keywords":["Mek inhibitors","Lc-ms/ms","Mhc-i","Ligandome analysis","Influenza a virus","Zapnometinib"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD041578"],"tissue":["Cell Culture"],"sample_protocol":["Direct infection of cells:  Calu-3 cells were infected with influenza A virus (IAV) strains A/Fukui/20/2004 (H3N2) at an MOI of 0.1. After 1 h incubation at 37 °C, the inocula were removed, cell monolayers washed with PBS++, and overlaid with infection media containing either the MEK inhibitor zapnometinib or DMSO as vehicle control.  At 24 hours post-infection (hpi), the cells were harvested using enzyme-free cell dissociation buffer (Gibco), washed twice times with cold PBS, and stored at – 80 °C. Isolation of naturally presented HLA ligands: HLA I ligands were isolated using standard immunoaffinity purification employing the pan-HLA class I specific W6/32 antibody (PMID: 23329485). HLA ligand extracts were analyzed as described previously (PMID: 23329485). The HLA ligands were separated by reverse-phase LC (UltiMate 3000 RSLCnano system, Dionex) using a 75 μm × 2 cm trapping column and subsequently a 25 cm length and 50 μm separation column (Thermo Fisher Scientific) applying a gradient ranging from 2.4 to 32 % AcN over 90 min in a flow rate of 175 nl/min. For the mass spectrometric analysis of HLA class I peptides, a top-speed collision-induced dissociation (CID) method generating ion trap MS/MS spectra in the mass range 400–650   m/z using an online coupled LTQ Orbitrap Fusion Lumos mass spectrometer (Thermo Fisher Scientific)."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["Data processing was performed as described previously (PMID: 25548167). In brief, data were processed against concatenated FASTA sequences containing the reviewed UniProt human and IAV proteome of the aforementioned strain (viewed January 2020) by applying the SequestHT algorithm in the Proteome Discoverer 1.4 (Thermo Fisher Scientific) software. Precursor mass tolerance was set to 5 ppm, fragment mass tolerance was set to 0.5 Da, and oxidized methionine was allowed as a dynamic modification. Percolator-assisted FDR algorithm (PMID: 17952086) was implemented at a target value of q ≤ 0.05 (5% FDR), search engine rank =1, and peptide lengths were limited to 8–12 amino acids."],"omics_type":["Proteomics"],"labhead":["Prof. Dr. Oliver Planz"],"instrument_platform":[""],"labhead_affiliation":["Department of Immunology, Interfaculty Institute for Cell Biology, University of Tübingen, Tübingen, Germany"],"submission_type":["PARTIAL"],"species":["Homo Sapiens (human)"],"submitter_mail":["hazem.ewess@mnf.uni-tuebingen.de"],"publication":["42273698 Hamza H, Ghosh M, Rammensee HG, Planz O. Zapnometinib treatment and influenza A virus infection modulate the HLA class I ligandome in human lung adenocarcinoma cells. Front Immunol. 2026 17:1790267 10.3389/fimmu.2026.1790267"],"submitter_affiliation":["Department of Immunology, Interfaculty Institute for Cell Biology, University of Tübingen"],"submitter_country":["Germany"],"pubmed_abstract":["<h4>Introduction</h4>Influenza viruses continue to pose a global health threat, and available antiviral therapies are limited by resistance and reduced efficacy. Host-directed drugs such as MEK inhibitors have emerged as promising alternatives. Zapnometinib, a clinical-stage MEK inhibitor, has shown both antiviral and immunomodulatory activity. However, its impact on antigen presentation at the level of the HLA-I ligandome has not been investigated.<h4>Methods</h4>Label-free LC-MS/MS was applied to analyze HLA-I-presented peptides in human lung adenocarcinoma Calu-3 cells. Cells were infected with Influenza A virus (IAV) H3N2/Fukui and treated with zapnometinib. Surface HLA-I expression was quantified by flow cytometry, and immunopeptidomic analyses were applied to assess ligandome alterations and functional pathway enrichment.<h4>Results</h4>Zapnometinib treatment and IAV infection did not significantly alter HLA-I surface expression (fold changes 1.01-1.13, p > 0.05). Immunopeptidomics revealed allotype-specific changes in the relative abundance of HLA-I-presented peptides (approximately 3-12% change per allotype), and statistically significant modulation of defined ligand subsets (about 3-14% of ligands per condition; log<sub>2</sub> fold change ≥ 2, adjusted p < 0.05). Functional annotation analysis showed condition-specific enrichment in distinct cellular pathways, such as interferon-induced pathways, including IFI16 upregulation during infection and its downregulation upon MEK inhibition, indicating selective modulation of antiviral responses, cell cycle and RNA-processing pathways under zapnometinib treatment, and adhesion-related pathways under combined treatment. Ligandome remodeling after IAV H3N2/Fukui infection strongly affected antiviral hubs (IFIH1, DHX58, IFI16), whereas zapnometinib caused no consistent or significant reduction.<h4>Discussion/conclusion</h4>This study provides the first evidence that zapnometinib and IAV H3N2/Fukui induce remarkable effects on the HLA-I ligandome plasticity without substantially affecting overall HLA-I expression. These findings highlight the dual role of MEK inhibitors in modulating both viral replication and immune recognition through pathway- and allotype-specific changes in peptide presentation rather than bulk HLA-I levels. Our results support further investigation of zapnometinib as a host-directed antiviral strategy with the potential to contribute to targeted immunomodulation in antiviral therapy."],"pubmed_title":["Zapnometinib treatment and influenza A virus infection modulate the HLA class I ligandome in human lung adenocarcinoma cells."],"pubmed_authors":["Hamza Hazem H, Ghosh Michael M, Rammensee Hans-Georg HG, Planz Oliver O"],"additional_accession":[]},"is_claimable":false,"name":"Zapnometinib treatment and influenza A virus infection induce alteration on the HLA class I ligandome of human lung adenocarcinoma cells","description":"Targeting cellular factors is considered a promising approach to combat viral infection and overcome the resistance issue associated with direct-acting antiviral (DAA) drugs administration. In the context of drug repurposing strategies, we have previously explored the antiviral efficacy of the MEK inhibitor zapnometinib against influenza virus and SARS-CoV-2. Herein, since zapnometinib was originally promoted as an anti-cancer drug, we aimed to investigate the impact of zapnometinib on the HLA class I ligandome post-treatment and IAV infection. By implementing the immunopeptidomics approach, the influence on the HLA-I allotype distribution of the HLA ligands was observed to coincide with alteration in the relative abundance of HLA-I-presented peptides. Furthermore, functional annotation analysis of the corresponding source proteins of presented ligands revealed a remarkable gene enrichment of distinct cellular processes. Taken together, zapnometinib and IAV showed a remarkable effect on the HLA-I ligandome plasticity as well as quantitative alterations in HLA ligandome composition.","dates":{"publication":"2026-06-15","submission":"2023-04-17"},"accession":"PXD041578","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:1073882","NEWT:111205","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:1590","NEWT:10034","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:1589","NEWT:135622","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:383379","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:317447","NEWT:4688","NEWT:111225","NEWT:7719","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:160621","NEWT:212790","NEWT:110368","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:1912919","NEWT:44688","NEWT:44689","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:44685","NEWT:161934","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:410661","NEWT:1140","NCBITaxon:2157","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:270643","NEWT:2065263","NEWT:177437","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:244366","NEWT:1246791","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:33548","NEWT:52299","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2709072","NEWT:2853422","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:294607","NEWT:1502","NEWT:128017","NEWT:376686","NEWT:95486","NEWT:1883446","NEWT:1233435","NEWT:109760","NEWT:29031","NEWT:235443","NEWT:1757312","NEWT:85694","NEWT:108458","NEWT:5936","NEWT:320637","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:1255228","NEWT:410289","NEWT:373153","NEWT:472","NEWT:940825","NEWT:529507","NEWT:1071661","NEWT:470","NEWT:5911","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:445","NEWT:153009","NEWT:261756","NEWT:63366","NEWT:63367","NEWT:215402","NEWT:1547","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:150475","NEWT:267872","NEWT:172269","NEWT:9534","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:2903","NEWT:185579","NEWT:13076","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:550","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:51338","NEWT:575412","NEWT:28112","NEWT:6493","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","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:8655","NEWT:5147","NEWT:28104","NEWT:407821","NCBITaxon:2","NEWT:568708","NEWT:986","NEWT:52641","NEWT:28532","NEWT:353152","NEWT:40674","NEWT:1194669","NEWT:51329","NEWT:443906","NEWT:519","NEWT:2510939","NEWT:7394","NEWT:6063","NEWT:1328388","NEWT:1548728","NEWT:667127","NEWT:9557","NEWT:377586","NEWT:300641","NEWT:39655","NEWT:9554","NEWT:38323","NEWT:256318","NEWT:206411","NCBITaxon:6191","NEWT:229533","NEWT:214053","NEWT:80863","NEWT:90675","NEWT:52638","NEWT:57075","NEWT:1666905","NEWT:8697","NEWT:8695","NEWT:884204","NEWT:1123869","NEWT:9544","NEWT:9545","NEWT:979","NEWT:7370","NEWT:83906","NEWT:1134506","NEWT:255470","NEWT:38783","NEWT:6426","NEWT:33090","NEWT:9935","NEWT:287889","NEWT:305959","NEWT:13489","NEWT:92867","NEWT:92866","NCBITaxon:3055","NEWT:51750","NEWT:202950","NEWT:295027","NCBITaxon:11320","NEWT:632957","NEWT:9925","NCBITaxon:9606","NEWT:90690","NEWT:1436183","NEWT:4232","NEWT:416348","NEWT:11298","NEWT:7742","NEWT:196627","NEWT:200308","NEWT:200302","NEWT:870435","NEWT:9913","NEWT:9915","NEWT:105841","NEWT:2666255","NEWT:999810","NCBITaxon:5693","NEWT:28995","NEWT:1392998","NEWT:380394","NEWT:226900","NEWT:1266738","NEWT:231490","NEWT:244704","NEWT:7725","NEWT:9909","NEWT:9902","NEWT:430615","NEWT:563041","NEWT:72664","NEWT:41949","NEWT:326423","NEWT:452467","NEWT:198822","NEWT:36111","NEWT:326424","NEWT:1678078","NEWT:749906","NEWT:418985","NEWT:749907","NE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