title: Targeted next-generation sequencing makes new molecular diagnoses and expands genotype-phenotype relationship in Ehlers-Danlos syndrome.
abstract: Ehlers-Danlos syndrome (EDS) comprises a group of overlapping hereditary disorders of connective tissue with significant morbidity and mortality, including major vascular complications. We sought to identify the diagnostic utility of a next-generation sequencing (NGS) panel in a mixed EDS cohort.
  
  
  
  We developed and applied PCR-based NGS assays for targeted, unbiased sequencing of 12 collagen and aortopathy genes to a cohort of 177 unrelated EDS patients. Variants were scored blind to previous genetic testing and then compared with results of previous Sanger sequencing.
  
  
  
  Twenty-eight pathogenic variants in COL5A1/2, COL3A1, FBN1, and COL1A1 and four likely pathogenic variants in COL1A1, TGFBR1/2, and SMAD3 were identified by the NGS assays. These included all previously detected single-nucleotide and other short pathogenic variants in these genes, and seven newly detected pathogenic or likely pathogenic variants leading to clinically significant diagnostic revisions. Twenty-two variants of uncertain significance were identified, seven of which were in aortopathy genes and required clinical follow-up.
  
  
  
  Unbiased NGS-based sequencing made new molecular diagnoses outside the expected EDS genotype-phenotype relationship and identified previously undetected clinically actionable variants in aortopathy susceptibility genes. These data may be of value in guiding future clinical pathways for genetic diagnosis in EDS.Genet Med 18 11, 1119-1127.
authors.affiliation: National Ehlers-Danlos Syndrome Diagnostic Service, Northwick Park Hospital, London, UK.|Department of Medicine, Institute of Clinical Sciences, Imperial College London, London, UK.|Vascular Unit, St Mary's Hospital, Imperial College London, London, UK.|Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK.
authors.forename: Ruwan A
authors.identifier: 0000-0002-6717-7383
authors.initials: RA
authors.lastname: Weerakkody
authors.affiliation: Department of Medicine, Institute of Clinical Sciences, Imperial College London, London, UK.
authors.forename: Jana
authors.identifier: 0000-0002-3664-3724
authors.initials: J
authors.lastname: Vandrovcova
authors.affiliation: Department of Medicine, Institute of Clinical Sciences, Imperial College London, London, UK.|Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK.|Department of Rheumatology, University College Hospital, London, UK.
authors.forename: Christina
authors.initials: C
authors.lastname: Kanonidou
authors.affiliation: Department of Medicine, Institute of Clinical Sciences, Imperial College London, London, UK.
authors.forename: Michael
authors.initials: M
authors.lastname: Mueller
authors.affiliation: Department of Medicine, Institute of Clinical Sciences, Imperial College London, London, UK.|Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK.
authors.forename: Piyush
authors.initials: P
authors.lastname: Gampawar
authors.affiliation: Department of Medicine, Institute of Clinical Sciences, Imperial College London, London, UK.
authors.forename: Yousef
authors.identifier: 0000-0001-9151-1241
authors.initials: Y
authors.lastname: Ibrahim
authors.affiliation: Department of Medicine, Institute of Clinical Sciences, Imperial College London, London, UK.
authors.forename: Penny
authors.identifier: 0000-0002-5572-8491
authors.initials: P
authors.lastname: Norsworthy
authors.affiliation: Department of Medicine, Institute of Clinical Sciences, Imperial College London, London, UK.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Biggs
authors.affiliation: Department of Medicine, Institute of Clinical Sciences, Imperial College London, London, UK.
authors.forename: Abdulshakur
authors.initials: A
authors.lastname: Abdullah
authors.affiliation: Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK.
authors.forename: David
authors.identifier: 0000-0002-6340-3742
authors.initials: D
authors.lastname: Ross
authors.affiliation: Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK.
authors.forename: Holly A
authors.initials: HA
authors.lastname: Black
authors.affiliation: Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, University of Oxford, Oxford, UK.
authors.forename: David
authors.initials: D
authors.lastname: Ferguson
authors.affiliation: Vascular Unit, St Mary's Hospital, Imperial College London, London, UK.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Cheshire
authors.affiliation: Department of Rheumatology, University College Hospital, London, UK.
authors.forename: Hanadi
authors.initials: H
authors.lastname: Kazkaz
authors.affiliation: Department of Rheumatology, University College Hospital, London, UK.
authors.forename: Rodney
authors.initials: R
authors.lastname: Grahame
authors.affiliation: National Ehlers-Danlos Syndrome Diagnostic Service, Northwick Park Hospital, London, UK.
authors.forename: Neeti
authors.initials: N
authors.lastname: Ghali
authors.affiliation: National Ehlers-Danlos Syndrome Diagnostic Service, Northwick Park Hospital, London, UK.|Current affiliation: Maritime Medical Genetics Service, IWK Health Centre & Dalhousie University, Halifax, Nova Scotia, Canada.
authors.forename: Anthony
authors.initials: A
authors.lastname: Vandersteen
authors.affiliation: National Ehlers-Danlos Syndrome Diagnostic Service, Northwick Park Hospital, London, UK.
authors.forename: F Michael
authors.initials: FM
authors.lastname: Pope
authors.affiliation: Department of Medicine, Institute of Clinical Sciences, Imperial College London, London, UK.|Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Aitman
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002648:Child; D002675:Child, Preschool; D003094:Collagen / Q000235:genetics*; D004535:Ehlers-Danlos Syndrome / Q000175:diagnosis* / Q000235:genetics* / Q000503:physiopathology; D005260:Female; D005820:Genetic Testing; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation / Q000235:genetics; D057089:Pathology, Molecular / Q000379:methods; D010641:Phenotype; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics; D055815:Young Adult
chemical_list: D018125:Receptors, Transforming Growth Factor beta; D003094:Collagen; D017346:Protein Serine-Threonine Kinases; D000077293:Receptor, Transforming Growth Factor-beta Type I; C000625841:TGFBR1 protein, human