A novel ceruloplasmin splicing mutation causes early-onset aceruloplasminemia with refractory microcytic anemia and iron overload

Published: 3 November 2025| Version 1 | DOI: 10.17632/5274tk6gk7.1
Contributor:
Xiao Liu

Description

We have provided the original data generated from whole-exome sequencing and Sanger sequencing of peripheral blood cells, as well as the predicted protein structure files of ceruloplasmin from a 9-year-old girl with aceruloplasminemia.

Files

Steps to reproduce

Whole exome sequencing Genomic DNA was extracted from the peripheral blood mononuclear cells of patients via a Qiagen DNA Mini Blood Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions. The Roche KAPA HyperExome probe set captured exons and adjacent splicing regions for sequencing on the MGISEQ-2000 platform. The raw data were processed and aligned to the hg19 reference genome via standard bioinformatics tools (SOAPnuke, BWA, and GATK). Variants were screened and annotated via public databases (ClinVar, OMIM, HGMD, 1000 Genomes Project, gnomAD, and ExAC) and classified according to ACMG/AMP guidelines. Sanger sequencing Mutations were analyzed via direct sequencing of all coding exons and exon‒intron boundaries of the ceruloplasmin (CP) gene, including at least 50 base pairs of flanking intronic sequences. PCR primers were designed via Primer3 version 0.4.0 (https://bioinfo.ut.ee/primer3-0.4.0/primer3, last accessed July 10, 2025) (Supplementary Table 1). The PCR products were purified and subjected to bidirectional Sanger sequencing. The sequencing results were aligned and compared to the reference sequence of CP (GenBank accession number NM_000096.3) and reviewed manually to identify potential pathogenic variants. Protein structure changes due to mutation were modeled via SWISS-MODEL. Protein structure visualization was performed via PyMol (v3.1.0a0 Open-Source, https://github.com/cgohlke/pymol-open-source-wheels/releases).

Institutions

Categories

Protein Structure, Exome Sequencing, Sanger Sequencing

Funders

  • Hunan Provincial Science and Technology Department
    Hunan, China
    Grant ID: 2025JJ80480
  • the Changsha Municipal Health Commission Scientific Research Program
    Grant ID: KJ-A2023006
  • The Affiliated Changsha Hospital of Xiangya School of Medicine, Central South University
    Grant ID: 2025CSSKKT74

Licence