Reliable Analysis of Clinical Tumor-Only Whole-Exome Sequencing Data

JCO Clin Cancer Inform. 2020 Apr;4:321-335. doi: 10.1200/CCI.19.00130.


Purpose: Allele-specific copy number alteration (CNA) analysis is essential to study the functional impact of single-nucleotide variants (SNVs) and the process of tumorigenesis. However, controversy over whether it can be performed with sufficient accuracy in data without matched normal profiles and a lack of open-source implementations have limited its application in clinical research and diagnosis.

Methods: We benchmark allele-specific CNA analysis performance of whole-exome sequencing (WES) data against gold standard whole-genome SNP6 microarray data and against WES data sets with matched normal samples. We provide a workflow based on the open-source PureCN R/Bioconductor package in conjunction with widely used variant-calling and copy number segmentation algorithms for allele-specific CNA analysis from WES without matched normals. This workflow further classifies SNVs by somatic status and then uses this information to infer somatic mutational signatures and tumor mutational burden (TMB).

Results: Application of our workflow to tumor-only WES data produces tumor purity and ploidy estimates that are highly concordant with estimates from SNP6 microarray data and matched normal WES data. The presence of cancer type-specific somatic mutational signatures was inferred with high accuracy. We also demonstrate high concordance of TMB between our tumor-only workflow and matched normal pipelines.

Conclusion: The proposed workflow provides, to our knowledge, the only open-source option with demonstrated high accuracy for comprehensive allele-specific CNA analysis and SNV classification of tumor-only WES. An implementation of the workflow is available on the Terra Cloud platform of the Broad Institute (Cambridge, MA).

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Algorithms*
  • Biomarkers, Tumor / genetics*
  • DNA Copy Number Variations*
  • Exome*
  • Gene Expression Regulation, Neoplastic
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Mutation*
  • Neoplasms / genetics*
  • Neoplasms / pathology
  • Neoplasms / therapy
  • Whole Exome Sequencing / methods*


  • Biomarkers, Tumor