title: Efficiency of exome sequencing for the molecular diagnosis of pseudoxanthoma elasticum.
abstract: The molecular etiology of pseudoxanthoma elasticum (PXE), an autosomal recessive connective tissue disorder, has become increasingly complex as not only mutations in ATP-binding cassette family C member 6 (ABCC6) but also ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) and gamma-glutamyl carboxylase (GGCX) can cause resembling phenotypes. Identification of modifier genes, such as vascular endothelial growth factor A, has further contributed to the molecular heterogeneity of PXE. In such heterogeneous diseases, next-generation sequencing (NGS) allows to perform mutation screening of several genes in a single reaction. We explored whole-exome sequencing (WES) as an efficient diagnostic tool to identify the causal mutations in ABCC6, GGCX, ENPP1, and vitamin K epoxide reductase complex, subunit 1 (VKORC1) in 16 PXE patients. WES identified a causal ABCC6 mutation in 30 out of 32 alleles and one GGCX mutation, whereas no causal mutations in ENPP1 or VKORC1 were detected. Exomes with insufficient reads (⩽20 depth) for the four genes and patients with single mutations were further evaluated by Sanger sequencing (SS), but no additional mutations were found. The potential of WES compared with targeted NGS is the ease to examine target genes and the opportunity to search for novel genes when targeted analysis is negative. Together with low cost, rapid and less laborious workflow, we conclude that WES complemented with SS can provide a tiered approach to molecular diagnostics of PXE.
authors.affiliation: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium; Department of Genetic Engineering and Biotechnology, Shahjalal University of Science and Technology, Sylhet, Bangladesh.
authors.forename: Mohammad J
authors.initials: MJ
authors.lastname: Hosen
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium.
authors.forename: Filip
authors.initials: F
authors.lastname: Van Nieuwerburgh
authors.affiliation: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors.forename: Wouter
authors.initials: W
authors.lastname: Steyaert
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium.
authors.forename: Dieter
authors.initials: D
authors.lastname: Deforce
authors.affiliation: Department of Dermatology, Angers University Hospital, L'UNAM University, Angers, France.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Martin
authors.affiliation: Department of Vascular Medicine, Angers University Hospital, L'UNAM University, Angers, France.
authors.forename: Georges
authors.initials: G
authors.lastname: Leftheriotis
authors.affiliation: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors.forename: Anne
authors.initials: A
authors.lastname: De Paepe
authors.affiliation: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Coucke
authors.affiliation: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium. Electronic address: olivier.vanakker@ugent.be.
authors.forename: Olivier M
authors.initials: OM
authors.lastname: Vanakker
mesh_terms: D000328:Adult; D000483:Alleles; D019735:Carbon-Carbon Ligases / Q000235:genetics; D059472:Exome; D005260:Female; D014644:Genetic Variation; D015894:Genome, Human; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D008875:Middle Aged; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics; D009154:Mutation; D057089:Pathology, Molecular; D010641:Phenotype; D010727:Phosphoric Diester Hydrolases / Q000235:genetics; D011561:Pseudoxanthoma Elasticum / Q000175:diagnosis* / Q000235:genetics*; D011755:Pyrophosphatases / Q000235:genetics; D017422:Sequence Analysis, DNA; D064417:Vitamin K Epoxide Reductases / Q000235:genetics
chemical_list: D019735:Carbon-Carbon Ligases; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D010727:Phosphoric Diester Hydrolases; D011755:Pyrophosphatases; D064417:Vitamin K Epoxide Reductases; C102029:ABCC6 protein, human; C578523:VKORC1 protein, human; C065962:ectonucleotide pyrophosphatase phosphodiesterase 1; C020660:glutamyl carboxylase