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In vitro proteasome processing of neo-splicetopes does not predict their presentation in vivo

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Item Type:Article
Title:In vitro proteasome processing of neo-splicetopes does not predict their presentation in vivo
Creators Name:Willimsky, G. and Beier, C. and Immisch, L. and Papafotiou, G. and Scheuplein-Schlosser, V. and Goede, A. and Holzhütter, H.G. and Blankenstein, T. and Kloetzel, P.M.
Abstract:Proteasome catalyzed peptide splicing (PCPS) of cancer-driving antigens could generate attractive neoepitopes to be targeted by TCR-based adoptive T cell therapy. Based on a spliced peptide prediction algorithm TCRs were generated against putative KRAS(G12V) and RAC2(P29L) derived neo-splicetopes with high HLA-A*02:01 binding affinity. TCRs generated in mice with a diverse human TCR repertoire specifically recognized the respective target peptides with high efficacy. However, we failed to detect any neo-splicetope specific T cell response when testing the in vivo neo-splicetope generation and obtained no experimental evidence that the putative KRAS(G12V)- and RAC2(P29L)-derived neo-splicetopes were naturally processed and presented. Furthermore, only the putative RAC2(P29L)-derived neo-splicetopes was generated by in vitro PCPS. The experiments pose severe questions on the notion that available algorithms or the in vitro PCPS reaction reliably simulate in vivo splicing and argue against the general applicability of an algorithm-driven 'reverse immunology' pipeline for the identification of cancer-specific neo-splicetopes.
Keywords:Antigen Presentation, Neoplasm Antigens, CD8-Positive T-Lymphocytes, Epitopes, HEK293 Cells, HLA-A2 Antigen, K562 Cells, Transgenic Mice, Mutation, Neoplasms, Proof of Concept Study, Proteasome Endopeptidase Complex, Post-Translational Protein Processing, Proto-Oncogene Proteins p21(ras), T-Cell Antigen Receptors, rac GTP-Binding Proteins, rac GTP-Binding Proteins / metabolism, Animals, Mice
Publisher:eLife Sciences Publications
Page Range:e62019
Date:20 April 2021
Official Publication:https://doi.org/10.7554/eLife.62019
PubMed:View item in PubMed

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