Structural Chromosomal Rearrangements Require Nucleotide-Level Resolution: Lessons from Next-Generation Sequencing in Prenatal Diagnosis

Am J Hum Genet. 2016 Nov 3;99(5):1015-1033. doi: 10.1016/j.ajhg.2016.08.022. Epub 2016 Oct 13.

Abstract

In this exciting era of "next-gen cytogenetics," integrating genomic sequencing into the prenatal diagnostic setting is possible within an actionable time frame and can provide precise delineation of balanced chromosomal rearrangements at the nucleotide level. Given the increased risk of congenital abnormalities in newborns with de novo balanced chromosomal rearrangements, comprehensive interpretation of breakpoints could substantially improve prediction of phenotypic outcomes and support perinatal medical care. Herein, we present and evaluate sequencing results of balanced chromosomal rearrangements in ten prenatal subjects with respect to the location of regulatory chromatin domains (topologically associated domains [TADs]). The genomic material from all subjects was interpreted to be "normal" by microarray analyses, and their rearrangements would not have been detected by cell-free DNA (cfDNA) screening. The findings of our systematic approach correlate with phenotypes of both pregnancies with untoward outcomes (5/10) and with healthy newborns (3/10). Two pregnancies, one with a chromosomal aberration predicted to be of unknown clinical significance and another one predicted to be likely benign, were terminated prior to phenotype-genotype correlation (2/10). We demonstrate that the clinical interpretation of structural rearrangements should not be limited to interruption, deletion, or duplication of specific genes and should also incorporate regulatory domains of the human genome with critical ramifications for the control of gene expression. As detailed in this study, our molecular approach to both detecting and interpreting the breakpoints of structural rearrangements yields unparalleled information in comparison to other commonly used first-tier diagnostic methods, such as non-invasive cfDNA screening and microarray analysis, to provide improved genetic counseling for phenotypic outcome in the prenatal setting.

MeSH terms

  • Alleles
  • Chromosome Aberrations*
  • Chromosome Mapping
  • Congenital Abnormalities / diagnosis
  • Congenital Abnormalities / genetics*
  • Female
  • Gene Expression Regulation
  • Gene Rearrangement*
  • Genetic Testing
  • Genome, Human
  • Genomics
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Karyotyping
  • Male
  • Nucleotides / genetics*
  • Pregnancy
  • Prenatal Diagnosis / methods*
  • SOX9 Transcription Factor / genetics
  • SOX9 Transcription Factor / metabolism
  • Sequence Analysis, DNA
  • Translocation, Genetic

Substances

  • Nucleotides
  • SOX9 Transcription Factor
  • SOX9 protein, human