title: Exploiting the mutanome for tumor vaccination.
abstract: Multiple genetic events and subsequent clonal evolution drive carcinogenesis, making disease elimination with single-targeted drugs difficult. The multiplicity of gene mutations derived from clonal heterogeneity therefore represents an ideal setting for multiepitope tumor vaccination. Here, we used next generation sequencing exome resequencing to identify 962 nonsynonymous somatic point mutations in B16F10 murine melanoma cells, with 563 of those mutations in expressed genes. Potential driver mutations occurred in classical tumor suppressor genes and genes involved in proto-oncogenic signaling pathways that control cell proliferation, adhesion, migration, and apoptosis. Aim1 and Trrap mutations known to be altered in human melanoma were included among those found. The immunogenicity and specificity of 50 validated mutations was determined by immunizing mice with long peptides encoding the mutated epitopes. One-third of these peptides were found to be immunogenic, with 60% in this group eliciting immune responses directed preferentially against the mutated sequence as compared with the wild-type sequence. In tumor transplant models, peptide immunization conferred in vivo tumor control in protective and therapeutic settings, thereby qualifying mutated epitopes that include single amino acid substitutions as effective vaccines. Together, our findings provide a comprehensive picture of the mutanome of B16F10 melanoma which is used widely in immunotherapy studies. In addition, they offer insight into the extent of the immunogenicity of nonsynonymous base substitution mutations. Lastly, they argue that the use of deep sequencing to systematically analyze immunogenicity mutations may pave the way for individualized immunotherapy of cancer patients.
authors.affiliation: TRON-Translational Oncology at the University Medical Center Mainz, Germany.
authors.forename: John C
authors.initials: JC
authors.lastname: Castle
authors.forename: Sebastian
authors.initials: S
authors.lastname: Kreiter
authors.forename: Jan
authors.initials: J
authors.lastname: Diekmann
authors.forename: Martin
authors.initials: M
authors.lastname: Löwer
authors.forename: Niels
authors.initials: N
authors.lastname: van de Roemer
authors.forename: Jos
authors.initials: J
authors.lastname: de Graaf
authors.forename: Abderraouf
authors.initials: A
authors.lastname: Selmi
authors.forename: Mustafa
authors.initials: M
authors.lastname: Diken
authors.forename: Sebastian
authors.initials: S
authors.lastname: Boegel
authors.forename: Claudia
authors.initials: C
authors.lastname: Paret
authors.forename: Michael
authors.initials: M
authors.lastname: Koslowski
authors.forename: Andreas N
authors.initials: AN
authors.lastname: Kuhn
authors.forename: Cedrik M
authors.initials: CM
authors.lastname: Britten
authors.forename: Christoph
authors.initials: C
authors.lastname: Huber
authors.forename: Ozlem
authors.initials: O
authors.lastname: Türeci
authors.forename: Ugur
authors.initials: U
authors.lastname: Sahin
mesh_terms: D000818:Animals; D019496:Cancer Vaccines / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D045744:Cell Line, Tumor; D000939:Epitopes / Q000235:genetics; D059472:Exome; D005260:Female; D008546:Melanoma, Experimental / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D051379:Mice; D008810:Mice, Inbred C57BL; D017354:Point Mutation / Q000276:immunology*; D017422:Sequence Analysis, DNA; D014611:Vaccination; D022223:Vaccines, Subunit / Q000627:therapeutic use
chemical_list: D019496:Cancer Vaccines; D000939:Epitopes; D022223:Vaccines, Subunit