title: Next-generation sequencing: Application of a novel platform to analyze atypical iron disorders.
abstract: The development of targeted next-generation sequencing (NGS) applications now promises to be a clinically viable option for the diagnosis of rare disorders. This approach is proving to have significant utility where standardized testing has failed to identify the underlying molecular basis of disease. We have developed a unique targeted NGS panel for the systematic sequence-based analysis of atypical iron disorders. We report the analysis of 39 genes associated with iron regulation in eight cases of atypical iron dysregulation, in which five cases we identified the definitive causative mutation, and a possible causative mutation in a sixth. We further provide a molecular and cellular characterization study of one of these mutations (TFR2, p.I529N) in a familial case as proof of principle. Cellular analysis of the mutant protein indicates that this amino acid substitution affects the localization of the protein, which results in its retention in the endoplasmic reticulum and thus failure to function at the cell surface. Our unique NGS panel presents a rapid and cost-efficient approach to identify the underlying genetic cause in cases of atypical iron homeostasis disorders.
authors.affiliation: Membrane Transport Laboratory, QIMR Berghofer Medical Research Institute, Australia.
authors.forename: Cameron J
authors.initials: CJ
authors.lastname: McDonald
authors.affiliation: Membrane Transport Laboratory, QIMR Berghofer Medical Research Institute, Australia.
authors.forename: Lesa
authors.initials: L
authors.lastname: Ostini
authors.affiliation: Membrane Transport Laboratory, QIMR Berghofer Medical Research Institute, Australia; Faculty of Medicine and Biomedical Sciences, The University of Queensland, Brisbane, Australia.
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Wallace
authors.affiliation: Gosford Hospital, Australia.
authors.forename: Alison
authors.initials: A
authors.lastname: Lyons
authors.affiliation: Faculty of Medicine and Biomedical Sciences, The University of Queensland, Brisbane, Australia.
authors.forename: Darrell H G
authors.initials: DH
authors.lastname: Crawford
authors.affiliation: Membrane Transport Laboratory, QIMR Berghofer Medical Research Institute, Australia; Faculty of Medicine and Biomedical Sciences, The University of Queensland, Brisbane, Australia. Electronic address: Nathan.Subramaniam@qimrberghofer.edu.au.
authors.forename: V Nathan
authors.initials: VN
authors.lastname: Subramaniam
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D004247:DNA / Q000235:genetics*; D004252:DNA Mutational Analysis; D005260:Female; D020022:Genetic Predisposition to Disease; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007501:Iron / Q000378:metabolism*; D019189:Iron Metabolism Disorders / Q000235:genetics* / Q000378:metabolism; D008875:Middle Aged; D009154:Mutation; D011990:Receptors, Transferrin / Q000235:genetics* / Q000378:metabolism
chemical_list: D011990:Receptors, Transferrin; D004247:DNA; D007501:Iron
keywords: Anemia; Hemochromatosis; Iron overload; Next-generation sequencing; Transferrin receptor 2