Integrative analysis of structural variations using short-reads and linked-reads yields highly specific and sensitive predictions

PLoS Comput Biol. 2020 Nov 23;16(11):e1008397. doi: 10.1371/journal.pcbi.1008397. eCollection 2020 Nov.

Abstract

Genetic diseases are driven by aberrations of the human genome. Identification of such aberrations including structural variations (SVs) is key to our understanding. Conventional short-reads whole genome sequencing (cWGS) can identify SVs to base-pair resolution, but utilizes only short-range information and suffers from high false discovery rate (FDR). Linked-reads sequencing (10XWGS) utilizes long-range information by linkage of short-reads originating from the same large DNA molecule. This can mitigate alignment-based artefacts especially in repetitive regions and should enable better prediction of SVs. However, an unbiased evaluation of this technology is not available. In this study, we performed a comprehensive analysis of different types and sizes of SVs predicted by both the technologies and validated with an independent PCR based approach. The SVs commonly identified by both the technologies were highly specific, while validation rate dropped for uncommon events. A particularly high FDR was observed for SVs only found by 10XWGS. To improve FDR and sensitivity, statistical models for both the technologies were trained. Using our approach, we characterized SVs from the MCF7 cell line and a primary breast cancer tumor with high precision. This approach improves SV prediction and can therefore help in understanding the underlying genetics in various diseases.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Breast Neoplasms / genetics
  • Computational Biology
  • DNA Barcoding, Taxonomic / methods
  • DNA Barcoding, Taxonomic / statistics & numerical data
  • DNA, Neoplasm / genetics
  • Female
  • Genetic Diseases, Inborn / diagnosis
  • Genetic Diseases, Inborn / genetics
  • Genome, Human
  • Genomic Structural Variation*
  • Genomics / methods
  • Genomics / statistics & numerical data
  • High-Throughput Nucleotide Sequencing / methods*
  • High-Throughput Nucleotide Sequencing / statistics & numerical data
  • Humans
  • Logistic Models
  • MCF-7 Cells
  • Polymerase Chain Reaction / methods
  • Polymerase Chain Reaction / statistics & numerical data
  • Sequence Analysis, DNA / methods
  • Sequence Analysis, DNA / statistics & numerical data
  • Whole Genome Sequencing / methods*
  • Whole Genome Sequencing / statistics & numerical data

Substances

  • DNA, Neoplasm

Grants and funding

This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No 789256) awarded to US. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.