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An analysis of the sensitivity of proteogenomic mapping of somatic mutations and novel splicing events in cancer

Item Type:Article
Title:An analysis of the sensitivity of proteogenomic mapping of somatic mutations and novel splicing events in cancer
Creators Name:Ruggles, K.V. and Tang, Z. and Wang, X. and Grover, H. and Askenazi, M. and Teubl, J. and Cao, S. and McLellan, M.D. and Clauser, K.R. and Tabb, D.L. and Mertins, P. and Slebos, R. and Erdmann-Gilmore, P. and Li, S. and Gunawardena, H.P. and Xie, L. and Liu, T. and Zhou, J.Y. and Sun, S. and Hoadley, K.A. and Perou, C.M. and Chen, X. and Davies, S.R. and Maher, C.A. and Kinsinger, C.R. and Rodland, K.D. and Zhang, H. and Zhang, Z. and Ding, L. and Townsend, R.R. and Rodriguez, H. and Chan, D. and Smith, R.D. and Liebler, D.C. and Carr, S.A. and Payne, S. and Ellis, M.J. and Fenyö, D.
Abstract:Improvements in mass spectrometry (MS)-based peptide sequencing provide a new opportunity to determine whether polymorphisms, mutations, and splice variants identified in cancer cells are translated. Herein, we apply a proteogenomic data integration tool (QUILTS) to illustrate protein variant discovery using whole genome, whole transcriptome, and global proteome datasets generated from a pair of luminal and basal-like breast-cancer-patient-derived xenografts (PDX). The sensitivity of proteogenomic analysis for singe nucleotide variant (SNV) expression and novel splice junction (NSJ) detection was probed using multiple MS/MS sample process replicates defined here as an independent tandem MS experiment using identical sample material. Despite analysis of over 30 sample process replicates, only about 10% of SNVs (somatic and germline) detected by both DNA and RNA sequencing were observed as peptides. An even smaller proportion of peptides corresponding to NSJ observed by RNA sequencing were detected (<0.1%). Peptides mapping to DNA-detected SNVs without a detectable mRNA transcript were also observed, suggesting that transcriptome coverage was incomplete (~80%). In contrast to germline variants, somatic variants were less likely to be detected at the peptide level in the basal-like tumor than in the luminal tumor, raising the possibility of differential translation or protein degradation effects. In conclusion, this large-scale proteogenomic integration allowed us to determine the degree to which mutations are translated and identify gaps in sequence coverage, thereby benchmarking current technology and progress toward whole cancer proteome and transcriptome analysis.
Keywords:Alternative Splicing, Computational Biology, DNA Sequence Analysis, Experimental Mammary Neoplasms, Genetic Databases, Genome, Mutation, Proteomics, RNA Sequence Analysis, Single Nucleotide Polymorphism, Tandem Mass Spectrometry, Transcriptome, Animals, Mice
Source:Molecular & Cellular Proteomics
Publisher:American Society for Biochemistry and Molecular Biology (U.S.A.)
Page Range:1060-1071
Date:1 March 2016
Official Publication:https://doi.org/10.1074/mcp.M115.056226
PubMed:View item in PubMed

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