Beyond the exome: utility of long-read whole genome sequencing in exome-negative autosomal recessive diseases

Genome Med. 2023 Dec 14;15(1):114. doi: 10.1186/s13073-023-01270-8.

Abstract

Background: Long-read whole genome sequencing (lrWGS) has the potential to address the technical limitations of exome sequencing in ways not possible by short-read WGS. However, its utility in autosomal recessive Mendelian diseases is largely unknown.

Methods: In a cohort of 34 families in which the suspected autosomal recessive diseases remained undiagnosed by exome sequencing, lrWGS was performed on the Pacific Bioscience Sequel IIe platform.

Results: Likely causal variants were identified in 13 (38%) of the cohort. These include (1) a homozygous splicing SV in TYMS as a novel candidate gene for lethal neonatal lactic acidosis, (2) a homozygous non-coding SV that we propose impacts STK25 expression and causes a novel neurodevelopmental disorder, (3) a compound heterozygous SV in RP1L1 with complex inheritance pattern in a family with inherited retinal disease, (4) homozygous deep intronic variants in LEMD2 and SNAP91 as novel candidate genes for neurodevelopmental disorders in two families, and (5) a promoter SNV in SLC4A4 causing non-syndromic band keratopathy. Surprisingly, we also encountered causal variants that could have been identified by short-read exome sequencing in 7 families. The latter highlight scenarios that are especially challenging at the interpretation level.

Conclusions: Our data highlight the continued need to address the interpretation challenges in parallel with efforts to improve the sequencing technology itself. We propose a path forward for the implementation of lrWGS sequencing in the setting of autosomal recessive diseases in a way that maximizes its utility.

Keywords: ABHD12; Autozygome; C1orf109; FLVCR1; Long-read sequencing; NID1; PKHD1; SHFM; STX3.

MeSH terms

  • Exome Sequencing
  • Exome*
  • Eye Proteins / genetics
  • Genes, Recessive
  • Humans
  • Infant, Newborn
  • Inheritance Patterns*
  • Intracellular Signaling Peptides and Proteins / genetics
  • Membrane Proteins / genetics
  • Mutation
  • Nuclear Proteins / genetics
  • Pedigree
  • Protein Serine-Threonine Kinases / genetics

Substances

  • RP1L1 protein, human
  • Eye Proteins
  • LEMD2 protein, human
  • Membrane Proteins
  • Nuclear Proteins
  • STK25 protein, human
  • Protein Serine-Threonine Kinases
  • Intracellular Signaling Peptides and Proteins