Anti-tumour immunity induces aberrant peptide presentation in melanoma

Tryptophan depletion in melanoma cells after prolonged treatment with interferon-gamma (IFN gamma) results in ribosomal frameshifting and the production of aberrant peptides that can be presented to T cells and induce an immune response. Extensive tumour inflammation, which is reflected by high levels of infiltrating T cells and interferon-gamma (IFN gamma) signalling, improves the response of patients with melanoma to checkpoint immunotherapy(1,2). Many tumours, however, escape by activating cellular pathways that lead to immunosuppression. One such mechanism is the production of tryptophan metabolites along the kynurenine pathway by the enzyme indoleamine 2,3-dioxygenase 1 (IDO1), which is induced by IFN gamma(3-5). However, clinical trials using inhibition of IDO1 in combination with blockade of the PD1 pathway in patients with melanoma did not improve the efficacy of treatment compared to PD1 pathway blockade alone(6,7), pointing to an incomplete understanding of the role of IDO1 and the consequent degradation of tryptophan in mRNA translation and cancer progression. Here we used ribosome profiling in melanoma cells to investigate the effects of prolonged IFN gamma treatment on mRNA translation. Notably, we observed accumulations of ribosomes downstream of tryptophan codons, along with their expected stalling at the tryptophan codon. This suggested that ribosomes bypass tryptophan codons in the absence of tryptophan. A detailed examination of these tryptophan-associated accumulations of ribosomes-which we term 'W-bumps'-showed that they were characterized by ribosomal frameshifting events. Consistently, reporter assays combined with proteomic and immunopeptidomic analyses demonstrated the induction of ribosomal frameshifting, and the generation and presentation of aberrant trans-frame peptides at the cell surface after treatment with IFN gamma. Priming of naive T cells from healthy donors with aberrant peptides induced peptide-specific T cells. Together, our results suggest that IDO1-mediated depletion of tryptophan, which is induced by IFN gamma, has a role in the immune recognition of melanoma cells by contributing to diversification of the peptidome landscape.