PMID- 35239002 OWN - NLM STAT- MEDLINE DCOM- 20220314 LR - 20220716 IS - 1420-9071 (Electronic) IS - 1420-682X (Print) IS - 1420-682X (Linking) VI - 79 IP - 3 DP - 2022 Mar 3 TI - Upstream open reading frames regulate translation of cancer-associated transcripts and encode HLA-presented immunogenic tumor antigens. PG - 171 LID - 10.1007/s00018-022-04145-0 [doi] LID - 171 AB - BACKGROUND: Upstream open reading frames (uORFs) represent translational control elements within eukaryotic transcript leader sequences. Recent data showed that uORFs can encode for biologically active proteins and human leukocyte antigen (HLA)-presented peptides in malignant and benign cells suggesting their potential role in cancer cell development and survival. However, the role of uORFs in translational regulation of cancer-associated transcripts as well as in cancer immune surveillance is still incompletely understood. METHODS: We examined the translational regulatory effect of 29 uORFs in 13 cancer-associated genes by dual-luciferase assays. Cellular expression and localization of uORF-encoded peptides (uPeptides) were investigated by immunoblotting and immunofluorescence-based microscopy. Furthermore, we utilized mass spectrometry-based immunopeptidome analyses in an extensive dataset of primary malignant and benign tissue samples for the identification of naturally presented uORF-derived HLA-presented peptides screening for more than 2000 uORFs. RESULTS: We provide experimental evidence for similarly effective translational regulation of cancer-associated transcripts through uORFs initiated by either canonical AUG codons or by alternative translation initiation sites (aTISs). We further demonstrate frequent cellular expression and reveal occasional specific cellular localization of uORF-derived peptides, suggesting uPeptide-specific biological implications. Immunopeptidome analyses delineated a set of 125 naturally presented uORF-derived HLA-presented peptides. Comparative immunopeptidome profiling of malignant and benign tissue-derived immunopeptidomes identified several tumor-associated uORF-derived HLA ligands capable to induce multifunctional T cell responses. CONCLUSION: Our data provide direct evidence for the frequent expression of uPeptides in benign and malignant human tissues, suggesting a potentially widespread function of uPeptides in cancer biology. These findings may inspire novel approaches in direct molecular as well as immunotherapeutic targeting of cancer-associated uORFs and uPeptides. CI - (c) 2022. The Author(s). FAU - Nelde, Annika AU - Nelde A AUID- ORCID: 0000-0001-8504-8481 AD - Clinical Collaboration Unit Translational Immunology, Department of Internal Medicine, German Cancer Consortium (DKTK), University Hospital Tubingen, Otfried-Muller-Str. 10, 72076, Tubingen, Germany. AD - Department of Immunology, Institute for Cell Biology, University of Tubingen, 72076, Tubingen, Germany. AD - Cluster of Excellence iFIT (EXC2180) "Image-Guided and Functionally Instructed Tumor Therapies", University of Tubingen, 72076, Tubingen, Germany. FAU - Flototto, Lea AU - Flototto L AD - Department of Medicine A, Hematology, Oncology, Hemostaseology and Pneumology, University Hospital Munster, Albert-Schweitzer-Campus 1A, 48149, Munster, Germany. FAU - Jurgens, Lara AU - Jurgens L AD - Department of Medicine A, Hematology, Oncology, Hemostaseology and Pneumology, University Hospital Munster, Albert-Schweitzer-Campus 1A, 48149, Munster, Germany. FAU - Szymik, Laura AU - Szymik L AD - Department of Medicine A, Hematology, Oncology, Hemostaseology and Pneumology, University Hospital Munster, Albert-Schweitzer-Campus 1A, 48149, Munster, Germany. FAU - Hubert, Elvira AU - Hubert E AD - Department of Medicine A, Hematology, Oncology, Hemostaseology and Pneumology, University Hospital Munster, Albert-Schweitzer-Campus 1A, 48149, Munster, Germany. FAU - Bauer, Jens AU - Bauer J AUID- ORCID: 0000-0003-3731-2385 AD - Clinical Collaboration Unit Translational Immunology, Department of Internal Medicine, German Cancer Consortium (DKTK), University Hospital Tubingen, Otfried-Muller-Str. 10, 72076, Tubingen, Germany. AD - Department of Immunology, Institute for Cell Biology, University of Tubingen, 72076, Tubingen, Germany. AD - Cluster of Excellence iFIT (EXC2180) "Image-Guided and Functionally Instructed Tumor Therapies", University of Tubingen, 72076, Tubingen, Germany. FAU - Schliemann, Christoph AU - Schliemann C AD - Department of Medicine A, Hematology, Oncology, Hemostaseology and Pneumology, University Hospital Munster, Albert-Schweitzer-Campus 1A, 48149, Munster, Germany. FAU - Kessler, Torsten AU - Kessler T AD - Department of Medicine A, Hematology, Oncology, Hemostaseology and Pneumology, University Hospital Munster, Albert-Schweitzer-Campus 1A, 48149, Munster, Germany. FAU - Lenz, Georg AU - Lenz G AD - Department of Medicine A, Hematology, Oncology, Hemostaseology and Pneumology, University Hospital Munster, Albert-Schweitzer-Campus 1A, 48149, Munster, Germany. FAU - Rammensee, Hans-Georg AU - Rammensee HG AD - Department of Immunology, Institute for Cell Biology, University of Tubingen, 72076, Tubingen, Germany. AD - Cluster of Excellence iFIT (EXC2180) "Image-Guided and Functionally Instructed Tumor Therapies", University of Tubingen, 72076, Tubingen, Germany. AD - German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), Partner Site Tubingen, 72076, Tubingen, Germany. FAU - Walz, Juliane S AU - Walz JS AUID- ORCID: 0000-0001-6404-7391 AD - Clinical Collaboration Unit Translational Immunology, Department of Internal Medicine, German Cancer Consortium (DKTK), University Hospital Tubingen, Otfried-Muller-Str. 10, 72076, Tubingen, Germany. juliane.walz@med.uni-tuebingen.de. AD - Department of Immunology, Institute for Cell Biology, University of Tubingen, 72076, Tubingen, Germany. juliane.walz@med.uni-tuebingen.de. AD - Cluster of Excellence iFIT (EXC2180) "Image-Guided and Functionally Instructed Tumor Therapies", University of Tubingen, 72076, Tubingen, Germany. juliane.walz@med.uni-tuebingen.de. AD - Dr. Margarete Fischer-Bosch Institute of Clinical Pharmacology, Robert Bosch Center for Tumor Diseases (RBCT), 70376, Stuttgart, Germany. juliane.walz@med.uni-tuebingen.de. FAU - Wethmar, Klaus AU - Wethmar K AD - Department of Medicine A, Hematology, Oncology, Hemostaseology and Pneumology, University Hospital Munster, Albert-Schweitzer-Campus 1A, 48149, Munster, Germany. klaus.wethmar@ukmuenster.de. LA - eng GR - 70113632/Deutsche Krebshilfe/ GR - E!11969 compare/Eurostars-2/ GR - WA 4608/1-2/Deutsche Forschungsgemeinschaft/ GR - EXC2180 390900677/Deutsche Forschungsgemeinschaft under Germany's Excellence Strategy/ GR - EXC2180 390900677/Deutsche Forschungsgemeinschaft under Germany's Excellence Strategy/ GR - 2016.177.2/Wilhelm Sander-Stiftung/ GR - 2016.177.3/Wilhelm Sander-Stiftung/ GR - DJCLS 05 R/2017/Jose Carreras Leukamie-Stiftung/ GR - 2451-0-0/Fortune Program of the University of Tubingen/ PT - Journal Article DEP - 20220303 PL - Switzerland TA - Cell Mol Life Sci JT - Cellular and molecular life sciences : CMLS JID - 9705402 RN - 0 (Antigens, Neoplasm) RN - 0 (Peptides) SB - IM MH - *Antigens, Neoplasm/genetics/metabolism MH - HEK293 Cells MH - Humans MH - Neoplasms/*genetics MH - Open Reading Frames MH - *Peptides/genetics/metabolism PMC - PMC8894207 OTO - NOTNLM OT - Cancer OT - Immunopeptidomics OT - Mass spectrometry OT - Translational control OT - uORF COIS- H.-G.R. is shareholder of Immatics Biotechnologies GmbH, Synimmune GmbH, and Curevac AG. The other authors declare no competing interests. EDAT- 2022/03/04 06:00 MHDA- 2022/03/15 06:00 PMCR- 2022/03/03 CRDT- 2022/03/03 12:18 PHST- 2021/10/07 00:00 [received] PHST- 2022/01/10 00:00 [accepted] PHST- 2021/12/21 00:00 [revised] PHST- 2022/03/03 12:18 [entrez] PHST- 2022/03/04 06:00 [pubmed] PHST- 2022/03/15 06:00 [medline] PHST- 2022/03/03 00:00 [pmc-release] AID - 10.1007/s00018-022-04145-0 [pii] AID - 4145 [pii] AID - 10.1007/s00018-022-04145-0 [doi] PST - epublish SO - Cell Mol Life Sci. 2022 Mar 3;79(3):171. doi: 10.1007/s00018-022-04145-0.