title: Strong spontaneous tumor neoantigen responses induced by a natural human carcinogen.
abstract: A key to improving cancer immunotherapy will be the identification of tumor-specific "neoantigens" that arise from mutations and augment the resultant host immune response. In this study we identified single nucleotide variants (SNVs) by RNA sequencing of asbestos-induced murine mesothelioma cell lines AB1 and AB1-HA. Using the NetMHCpan 2.8 algorithm, the theoretical binding affinity of predicted peptides arising from high-confidence, exonic, non-synonymous SNVs was determined for the BALB/c strain. The immunoreactivity to 20 candidate mutation-carrying peptides of increased affinity and the corresponding wild-type peptides was determined using interferon-γ ELISPOT assays and lymphoid organs of non-manipulated tumor-bearing mice. A strong endogenous immune response was demonstrated to one of the candidate neoantigens, Uqcrc2; this response was detected in the draining lymph node and spleen. Antigen reactive cells were not detected in non-tumor bearing mice. The magnitude of the response to the Uqcrc2 neoantigen was similar to that of the strong influenza hemagglutinin antigen, a model tumor neoantigen. This work confirms that the approach of RNAseq plus peptide prediction and ELISPOT testing is sufficient to identify natural tumor neoantigens.
authors.affiliation: National Centre for Asbestos Related Diseases; School of Medicine and Pharmacology; University of Western Australia ; Nedlands, Australia ; Australian Mesothelioma Tumour Bank; Sir Charles Gairdner Hospital ; Nedlands, Australia.
authors.forename: Jenette
authors.initials: J
authors.lastname: Creaney
authors.affiliation: National Centre for Asbestos Related Diseases; School of Medicine and Pharmacology; University of Western Australia ; Nedlands, Australia.
authors.forename: Shaokang
authors.initials: S
authors.lastname: Ma
authors.affiliation: National Centre for Asbestos Related Diseases; School of Medicine and Pharmacology; University of Western Australia ; Nedlands, Australia.
authors.forename: Sophie A
authors.initials: SA
authors.lastname: Sneddon
authors.affiliation: National Centre for Asbestos Related Diseases; School of Medicine and Pharmacology; University of Western Australia ; Nedlands, Australia.
authors.forename: Michelle R
authors.initials: MR
authors.lastname: Tourigny
authors.affiliation: National Centre for Asbestos Related Diseases; School of Medicine and Pharmacology; University of Western Australia ; Nedlands, Australia ; Australian Mesothelioma Tumour Bank; Sir Charles Gairdner Hospital ; Nedlands, Australia.
authors.forename: Ian M
authors.initials: IM
authors.lastname: Dick
authors.affiliation: National Centre for Asbestos Related Diseases; School of Medicine and Pharmacology; University of Western Australia ; Nedlands, Australia ; Australian Mesothelioma Tumour Bank; Sir Charles Gairdner Hospital ; Nedlands, Australia.
authors.forename: Justine S
authors.initials: JS
authors.lastname: Leon
authors.affiliation: National Centre for Asbestos Related Diseases; School of Medicine and Pharmacology; University of Western Australia ; Nedlands, Australia.
authors.forename: Andrea
authors.initials: A
authors.lastname: Khong
authors.affiliation: National Centre for Asbestos Related Diseases; School of Medicine and Pharmacology; University of Western Australia ; Nedlands, Australia.
authors.forename: Scott A
authors.initials: SA
authors.lastname: Fisher
authors.affiliation: National Centre for Asbestos Related Diseases; School of Medicine and Pharmacology; University of Western Australia ; Nedlands, Australia.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Lake
authors.affiliation: National Centre for Asbestos Related Diseases; School of Medicine and Pharmacology; University of Western Australia ; Nedlands, Australia.
authors.forename: W Joost
authors.initials: WJ
authors.lastname: Lesterhuis
authors.affiliation: National Centre for Asbestos Related Diseases; School of Medicine and Pharmacology; University of Western Australia ; Nedlands, Australia ; Department of Oncology; Sir Charles Gairdner Hospital ; Nedlands, Australia.
authors.forename: Anna K
authors.initials: AK
authors.lastname: Nowak
authors.affiliation: Institute for Immunology and Infectious Diseases; Murdoch University ; Murdoch, Australia.
authors.forename: Shay
authors.initials: S
authors.lastname: Leary
authors.affiliation: Institute for Immunology and Infectious Diseases; Murdoch University ; Murdoch, Australia.
authors.forename: Mark W
authors.initials: MW
authors.lastname: Watson
authors.affiliation: National Centre for Asbestos Related Diseases; School of Medicine and Pharmacology; University of Western Australia ; Nedlands, Australia ; Australian Mesothelioma Tumour Bank; Sir Charles Gairdner Hospital ; Nedlands, Australia ; Department of Respiratory Medicine; Sir Charles Gairdner Hospital ; Nedlands, Australia.
authors.forename: Bruce W
authors.initials: BW
authors.lastname: Robinson
keywords: HA, hemagglutinin; MCA, methylcholanthrene; NGS, next-generation sequencing; RNAseq; SNV, single nucleotide variant.; dLN, lymph node draining the tumor; epitope prediction; mesothelioma; neoantigen