Utilize este identificador para referenciar este registo: http://hdl.handle.net/10400.18/3821
Título: Next-generation sequencing of iron-metabolism related genes in Portuguese patients with iron overload: novel pathogenic genetic variants
Autor: Faria, Ricardo
Silva, Bruno
Silva, Catarina
Vieira, Luis
Loureiro, Pedro
Gomes, Susana
Gonçalves, João
Rivera, Isabel
Fraga, Sofia
Fleming, Rita
Faustino, Paula
Palavras-chave: Next-generation Sequencing
Metabolismo do Ferro
Novas Variantes Genéticas
Doenças Genéticas
Data: Abr-2016
Resumo: Objective: In Southern European countries up to one-third of the patients with hereditary hemochromatosis (HH) do not present the common HFE risk genotype. In order to investigate the molecular basis of these cases we have designed a gene panel for rapid and simultaneous analysis of 6 HH-related genes (HFE, TFR2, HJV, HAMP, SLC40A1 and FTL) by next-generation sequencing (NGS). Materials and Methods: Eighty-eight iron overload Portuguese patients, negative for the common HFE mutations, were analysed. A TruSeq Custom Amplicon kit (TSCA, by Illumina) was designed in order to generate 97 amplicons covering exons, intron/exon junctions and UTRs of the mentioned genes with a cumulative target sequence of 12115bp. Amplicons were sequenced in the MiSeq instrument (IIlumina) using 250bp paired-end reads. Sequences were aligned against human genome reference hg19 using alignment and variant caller algorithms in the MiSeq reporter software. Novel variants were validated by Sanger sequencing and their pathogenic significance were assessed by in silico studies. Results: We found a total of 55 different genetic variants. These include novel pathogenic missense and splicing variants (in HFE and TFR2), a very rare variant in IRE of FTL, a variant that originates a novel translation initiation codon in the HAMP gene, among others. Conclusion: The merging of TSCA methodology and NGS technology appears to be an appropriate tool for simultaneous and fast analysis of HH-related genes in a large number of samples. However, establishing the clinical relevance of NGS-detected variants for HH development remains a hard-working task, requiring further functional studies.
Peer review: yes
URI: http://hdl.handle.net/10400.18/3821
Aparece nas colecções:DGH - Posters/abstracts em congressos internacionais

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