title: Next generation sequencing in a large cohort of patients presenting with neuromuscular disease before or at birth.
abstract: BACKGROUND
  Fetal akinesia/hypokinesia, arthrogryposis and severe congenital myopathies are heterogeneous conditions usually presenting before or at birth. Although numerous causative genes have been identified for each of these disease groups, in many cases a specific genetic diagnosis remains elusive. Due to the emergence of next generation sequencing, virtually the entire coding region of an individual's DNA can now be analysed through "whole" exome sequencing, enabling almost all known and novel disease genes to be investigated for disorders such as these.
  
  
  METHODS
  Genomic DNA samples from 45 patients with fetal akinesia/hypokinesia, arthrogryposis or severe congenital myopathies from 38 unrelated families were subjected to next generation sequencing. Clinical features and diagnoses for each patient were supplied by referring clinicians. Genomic DNA was used for either whole exome sequencing or a custom-designed neuromuscular sub-exomic supercapture array containing 277 genes responsible for various neuromuscular diseases. Candidate disease-causing variants were investigated and confirmed using Sanger sequencing. Some of the cases within this cohort study have been published previously as separate studies.
  
  
  RESULTS
  A conclusive genetic diagnosis was achieved for 18 of the 38 families. Within this cohort, mutations were found in eight previously known neuromuscular disease genes (CHRND, CHNRG, ECEL1, GBE1, MTM1, MYH3, NEB and RYR1) and four novel neuromuscular disease genes were identified and have been published as separate reports (GPR126, KLHL40, KLHL41 and SPEG). In addition, novel mutations were identified in CHRND, KLHL40, NEB and RYR1. Autosomal dominant, autosomal recessive, X-linked, and de novo modes of inheritance were observed.
  
  
  CONCLUSIONS
  By using next generation sequencing on a cohort of 38 unrelated families with fetal akinesia/hypokinesia, arthrogryposis, or severe congenital myopathy we therefore obtained a genetic diagnosis for 47% of families. This study highlights the power and capacity of next generation sequencing (i) to determine the aetiology of genetically heterogeneous neuromuscular diseases, (ii) to identify novel disease genes in small pedigrees or isolated cases and (iii) to refine the interplay between genetic diagnosis and clinical evaluation and management.
authors.affiliation: Harry Perkins Institute of Medical Research and the Centre for Medical Research, University of Western Australia, QQ Block, 6 Verdun Street, Nedlands, 6009, , WA, Australia. emilytodd@live.com.
authors.forename: Emily J
authors.initials: EJ
authors.lastname: Todd
authors.affiliation: Harry Perkins Institute of Medical Research and the Centre for Medical Research, University of Western Australia, QQ Block, 6 Verdun Street, Nedlands, 6009, , WA, Australia. 20163622@student.uwa.edu.au.
authors.forename: Kyle S
authors.initials: KS
authors.lastname: Yau
authors.affiliation: Harry Perkins Institute of Medical Research and the Centre for Medical Research, University of Western Australia, QQ Block, 6 Verdun Street, Nedlands, 6009, , WA, Australia. royston.ong@uwa.edu.au.
authors.forename: Royston
authors.initials: R
authors.lastname: Ong
authors.affiliation: Genetic Services of Western Australia, King Edward Memorial Hospital, Perth, 6000, , WA, Australia. jennie.slee@health.wa.gov.au.
authors.forename: Jennie
authors.initials: J
authors.lastname: Slee
authors.affiliation: Victorian Clinical Genetics Services, Murdoch Children's Research Institute, The Royal Children's Hospital, Parkville, 3052, , VIC, Australia. george.mcgillivray@vcgs.org.au.
authors.forename: George
authors.initials: G
authors.lastname: McGillivray
authors.affiliation: Paediatric and Reproductive Genetics Unit, South Australia Clinical Genetics Service, Women's and Children's Hospital, North Adelaide, 5006, , SA, Australia. christopher.barnett@health.sa.gov.au.
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Barnett
authors.affiliation: Department of Pediatric Neurology, Hacettepe University Children's Hospital, Ankara, 06100, Turkey. gtuncer@hacettepe.edu.tr.
authors.forename: Goknur
authors.initials: G
authors.lastname: Haliloglu
authors.affiliation: Pediatric Pathology Unit, Hacettepe University Children's Hospital, Ankara, 06100, Turkey. btalim@hacettepe.edu.tr.
authors.forename: Beril
authors.initials: B
authors.lastname: Talim
authors.affiliation: Pediatric Pathology Unit, Hacettepe University Children's Hospital, Ankara, 06100, Turkey. zakcoren@hacettepe.edu.tr.
authors.forename: Zuhal
authors.initials: Z
authors.lastname: Akcoren
authors.affiliation: Kariminejad-Najmabadi Pathology and Genetics Centre, Tehran, 14656, Iran. arianakariminejad@yahoo.com.
authors.forename: Ariana
authors.initials: A
authors.lastname: Kariminejad
authors.affiliation: Royal Children's Hospital, Herston Road, Herson, 4029, , QLD, Australia. Anita_Cairns@health.qld.gov.au.
authors.forename: Anita
authors.initials: A
authors.lastname: Cairns
authors.affiliation: Institute for Neuroscience and Muscle Research, The Children's Hospital at Westmead, Sydney, 2145, , NSW, Australia. nigel.clarke@health.nsw.gov.au.|Discipline of Paediatrics and Child Health, University of Sydney, Sydney, 2006, , NSW, Australia. nigel.clarke@health.nsw.gov.au.
authors.forename: Nigel F
authors.initials: NF
authors.lastname: Clarke
authors.affiliation: Sydney Children's Hospital, High Street, Randwick, 2031, , NSW, Australia. Mary-Louise.Freckmann@sesiahs.health.nsw.gov.au.
authors.forename: Mary-Louise
authors.initials: ML
authors.lastname: Freckmann
authors.affiliation: Unitè de Morphologie Neuromusculaire, Institut de Myologie, Institut National de la Santè et de la Recherche Mèdicale, Paris, 75651, France. nb.romero@institut-myologie.org.
authors.forename: Norma B
authors.initials: NB
authors.lastname: Romero
authors.affiliation: Dubowitz Neuromuscular Centre, UCL Institute of Child Health, London, WC1N 1EH, UK. denise.williams@bwnft.nhs.uk.|Wolfson Centre for Neuromuscular Disorders, RJAH Orthopaedic Hospital, Oswestry, SY10 7AG, UK. denise.williams@bwnft.nhs.uk.
authors.forename: Denise
authors.initials: D
authors.lastname: Williams
authors.affiliation: Dubowitz Neuromuscular Centre, UCL Institute of Child Health, London, WC1N 1EH, UK. c.sewry@imperial.ac.uk.|Wolfson Centre for Neuromuscular Disorders, RJAH Orthopaedic Hospital, Oswestry, SY10 7AG, UK. c.sewry@imperial.ac.uk.
authors.forename: Caroline A
authors.initials: CA
authors.lastname: Sewry
authors.affiliation: Department of Clinical Genetics, South Western Sydney Local Health District, Liverpool, 1871, , NSW, Australia. alison.colley@sswahs.nsw.gov.au.
authors.forename: Alison
authors.initials: A
authors.lastname: Colley
authors.affiliation: Department of Neurology, The Royal Children's Hospital, Melbourne, 3000, , VIC, Australia. monique.ryan@rch.org.au.
authors.forename: Monique M
authors.initials: MM
authors.lastname: Ryan
authors.affiliation: Genetic Services of Western Australia, Princess Margaret Hospital for Children and King Edward Memorial Hospital for Women, Subiaco, 6008, , WA, Australia. Cathy.Kiraly-Borri@health.wa.gov.au.
authors.forename: Cathy
authors.initials: C
authors.lastname: Kiraly-Borri
authors.affiliation: Department of Diagnostic Genomics, Pathwest, QEII Medical Centre, Nedlands, 6009, , WA, Australia. padma.sivadorai@health.wa.gov.au.
authors.forename: Padma
authors.initials: P
authors.lastname: Sivadorai
authors.affiliation: Lotterywest State Biomedical Facility Genomics and School of Pathology and Laboratory Medicine, University of Western Australia, Perth, 6000, , WA, Australia. richard.allcock@uwa.edu.au.
authors.forename: Richard J N
authors.initials: RJ
authors.lastname: Allcock
authors.affiliation: Nuffield Department of Clinical Neurosciences, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS, UK. david.beeson@ndcn.ox.ac.uk.
authors.forename: David
authors.initials: D
authors.lastname: Beeson
authors.affiliation: Nuffield Department of Clinical Neurosciences, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS, UK. susan.maxwell@imm.ox.ac.uk.
authors.forename: Susan
authors.initials: S
authors.lastname: Maxwell
authors.affiliation: Department of Diagnostic Genomics, Pathwest, QEII Medical Centre, Nedlands, 6009, , WA, Australia. mark.davis@health.wa.gov.au.
authors.forename: Mark R
authors.initials: MR
authors.lastname: Davis
authors.affiliation: Harry Perkins Institute of Medical Research and the Centre for Medical Research, University of Western Australia, QQ Block, 6 Verdun Street, Nedlands, 6009, , WA, Australia. nigel.laing@uwa.edu.au.|Department of Diagnostic Genomics, Pathwest, QEII Medical Centre, Nedlands, 6009, , WA, Australia. nigel.laing@uwa.edu.au.
authors.forename: Nigel G
authors.initials: NG
authors.lastname: Laing
authors.affiliation: Harry Perkins Institute of Medical Research and the Centre for Medical Research, University of Western Australia, QQ Block, 6 Verdun Street, Nedlands, 6009, , WA, Australia. gina.ravenscroft@perkins.uwa.edu.au.
authors.forename: Gianina
authors.initials: G
authors.lastname: Ravenscroft
mesh_terms: D000595:Amino Acid Sequence; D002648:Child; D002675:Child, Preschool; D015331:Cohort Studies; D005260:Female; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods* / Q000639:trends; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008969:Molecular Sequence Data; D009468:Neuromuscular Diseases / Q000175:diagnosis* / Q000235:genetics*; D010375:Pedigree; D011296:Prenatal Diagnosis / Q000379:methods* / Q000639:trends