BRCA1-associated R-loop affects transcription and differentiation in breast luminal epithelial cells

Nucleic Acids Res. 2019 Jun 4;47(10):5086-5099. doi: 10.1093/nar/gkz262.

Abstract

BRCA1-associated basal-like breast cancer originates from luminal progenitor cells. Breast epithelial cells from cancer-free BRCA1 mutation carriers are defective in luminal differentiation. However, how BRCA1 deficiency leads to lineage-specific differentiation defect is not clear. BRCA1 is implicated in resolving R-loops, DNA-RNA hybrid structures associated with genome instability and transcriptional regulation. We recently showed that R-loops are preferentially accumulated in breast luminal epithelial cells of BRCA1 mutation carriers. Here, we interrogate the impact of a BRCA1 mutation-associated R-loop located in a putative transcriptional enhancer upstream of the ERα-encoding ESR1 gene. Genetic ablation confirms the relevance of this R-loop-containing region to enhancer-promoter interactions and transcriptional activation of the corresponding neighboring genes, including ESR1, CCDC170 and RMND1. BRCA1 knockdown in ERα+ luminal breast cancer cells increases intensity of this R-loop and reduces transcription of its neighboring genes. The deleterious effect of BRCA1 depletion on transcription is mitigated by ectopic expression of R-loop-removing RNase H1. Furthermore, RNase H1 overexpression in primary breast cells from BRCA1 mutation carriers results in a shift from luminal progenitor cells to mature luminal cells. Our findings suggest that BRCA1-dependent R-loop mitigation contributes to luminal cell-specific transcription and differentiation, which could in turn suppress BRCA1-associated tumorigenesis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • BRCA1 Protein / genetics*
  • BRCA1 Protein / metabolism
  • Breast / metabolism*
  • CRISPR-Cas Systems
  • Carcinogenesis
  • Cell Differentiation
  • Enhancer Elements, Genetic*
  • Epithelial Cells / metabolism*
  • Estrogen Receptor alpha / genetics
  • Female
  • Gene Deletion
  • Gene Expression Regulation, Neoplastic*
  • Genes, BRCA1
  • HEK293 Cells
  • Heterozygote
  • Humans
  • MCF-7 Cells
  • Mutation
  • Transcription, Genetic

Substances

  • BRCA1 Protein
  • BRCA1 protein, human
  • ESR1 protein, human
  • Estrogen Receptor alpha