Identification and Functional Testing of ERCC2 Mutations in a Multi-national Cohort of Patients with Familial Breast- and Ovarian Cancer

PLoS Genet. 2016 Aug 9;12(8):e1006248. doi: 10.1371/journal.pgen.1006248. eCollection 2016 Aug.

Abstract

The increasing application of gene panels for familial cancer susceptibility disorders will probably lead to an increased proposal of susceptibility gene candidates. Using ERCC2 DNA repair gene as an example, we show that proof of a possible role in cancer susceptibility requires a detailed dissection and characterization of the underlying mutations for genes with diverse cellular functions (in this case mainly DNA repair and basic cellular transcription). In case of ERCC2, panel sequencing of 1345 index cases from 587 German, 405 Lithuanian and 353 Czech families with breast and ovarian cancer (BC/OC) predisposition revealed 25 mutations (3 frameshift, 2 splice-affecting, 20 missense), all absent or very rare in the ExAC database. While 16 mutations were unique, 9 mutations showed up repeatedly with population-specific appearance. Ten out of eleven mutations that were tested exemplarily in cell-based functional assays exert diminished excision repair efficiency and/or decreased transcriptional activation capability. In order to provide evidence for BC/OC predisposition, we performed familial segregation analyses and screened ethnically matching controls. However, unlike the recently published RECQL example, none of our recurrent ERCC2 mutations showed convincing co-segregation with BC/OC or significant overrepresentation in the BC/OC cohort. Interestingly, we detected that some deleterious founder mutations had an unexpectedly high frequency of > 1% in the corresponding populations, suggesting that either homozygous carriers are not clinically recognized or homozygosity for these mutations is embryonically lethal. In conclusion, we provide a useful resource on the mutational landscape of ERCC2 mutations in hereditary BC/OC patients and, as our key finding, we demonstrate the complexity of correct interpretation for the discovery of "bonafide" breast cancer susceptibility genes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Breast Neoplasms / genetics*
  • Breast Neoplasms / pathology
  • DNA Repair / genetics
  • Female
  • Genetic Predisposition to Disease*
  • Germ-Line Mutation
  • Heterozygote
  • Humans
  • Mutation, Missense
  • Ovarian Neoplasms / genetics*
  • Ovarian Neoplasms / pathology
  • Xeroderma Pigmentosum Group D Protein / chemistry
  • Xeroderma Pigmentosum Group D Protein / genetics*

Substances

  • Xeroderma Pigmentosum Group D Protein
  • ERCC2 protein, human

Supplementary concepts

  • Breast Cancer, Familial

Grants and funding

The Czech-Brno study was supported by MH CZ - DRO (MMCI, 00209805) and RECAMO, CZ.1.05/2.1.00/03.0101. The Czech-Prague study was supported by Ministry of Health of the Czech Republic, grant nr. 15-28830A and projects of the Charles University in Prague PRVOUK P27 and SVV UK 260256/2016. Functional analyses were funded by the Claudia von Schilling Foundation for breast cancer research (to SS and SE). The Lithuanian study was supported by the Research Counsil of Lithuania (Project SEN18/2015). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.