Identification of Novel Mutations by Targeted NGS Panel in Patients with Hyperferritinemia

Genes (Basel). 2021 Nov 9;12(11):1778. doi: 10.3390/genes12111778.

Abstract

Background: Several inherited diseases cause hyperferritinemia with or without iron overload. Differential diagnosis is complex and requires an extensive work-up. Currently, a clinical-guided approach to genetic tests is performed based on gene-by-gene sequencing. Although reasonable, this approach is expensive and time-consuming and Next Generation Sequencing (NGS) technology may provide cheaper and quicker large-scale DNA sequencing.

Methods: We analysed 36 patients with non-HFE-related hyperferritinemia. Liver iron concentration was measured in 33 by magnetic resonance. A panel of 25 iron related genes was designed using SureDesign software. Custom libraries were generated and then sequenced using Ion Torrent PGM.

Results: We identified six novel mutations in SLC40A1, three novel and one known mutation in TFR2, one known mutation and a de-novo deletion in HJV, and a novel mutation in HAMP in ten patients. In silico analyses supported the pathogenic role of the mutations.

Conclusions: Our results support the use of an NGS-based panel in selected patients with hyperferritinemia in a tertiary center for iron metabolism disorders. However, 26 out of 36 patients did not show genetic variants that can individually explain hyperferritinemia and/or iron overload suggesting the existence of other genetic defects or gene-gene and gene-environment interactions needing further studies.

Keywords: ferritin; ferroportin; hemochromatosis; hemojuvelin; hepcidin; iron overload; next generation sequencing; transferrin receptor 2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Cation Transport Proteins / genetics*
  • Child
  • Female
  • GPI-Linked Proteins / genetics*
  • Gene-Environment Interaction
  • Hemochromatosis Protein / genetics*
  • Hepcidins / genetics*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Hyperferritinemia / genetics*
  • Male
  • Middle Aged
  • Mutation*
  • Receptors, Transferrin / genetics*
  • Sequence Analysis, DNA / methods*
  • Tertiary Care Centers
  • Young Adult

Substances

  • Cation Transport Proteins
  • GPI-Linked Proteins
  • HAMP protein, human
  • HJV protein, human
  • Hemochromatosis Protein
  • Hepcidins
  • Receptors, Transferrin
  • TFR2 protein, human
  • metal transporting protein 1