Identification of a novel ANK1 gene variant c.1504-9G>A and its mechanism of intron retention in hereditary spherocytosis

Ting Xiong; Zhongjin Xu; Qian Wan; Feng Chen; Yao Ye; Hong Wang; Chongjun Wu

doi:10.3389/fgene.2024.1390924

Identification of a novel ANK1 gene variant c.1504-9G>A and its mechanism of intron retention in hereditary spherocytosis

Front Genet. 2024 Apr 9:15:1390924. doi: 10.3389/fgene.2024.1390924. eCollection 2024.

Authors

Ting Xiong¹, Zhongjin Xu², Qian Wan², Feng Chen², Yao Ye², Hong Wang², Chongjun Wu²

Affiliations

¹ Department of Endocrine Genetics and Metabolism, Jiangxi Provincial Children's Hospital, Nanchang, China.
² Department of Hematology, Jiangxi Provincial Children's Hospital, Nanchang, China.

Abstract

Objective: The objective of this study was to pinpoint pathogenic genes and assess the mutagenic pathogenicity in two pediatric patients with hereditary spherocytosis. Methods: We utilized whole-exome sequencing (WES) for individual analysis (case 1) and family-based trio analysis (case 2). The significance of the intronic mutation was validated through a Minigene splicing assay and supported by subsequent in vitro experiments. Results: Both probands received a diagnosis of hereditary spherocytosis. WES identified a novel ANK1 c.1504-9G>A mutation in both patients, causing the retention of seven nucleotides at the 5' end of intron 13, as substantiated by the Minigene assay. This variant results in a premature stop codon and the production of a truncated protein. In vitro studies indicated a reduced expression of the ANK1 gene. Conclusion: The novel ANK1 c.1504-9G>A variant is established as the causative factor for hereditary spherocytosis, with the c.1504-9G site functioning as a splicing receptor.

Keywords: ANK1; hereditary spherocytosis; intron retention; minigene splicing assay; whole-exome sequencing.

Grants and funding

The authors declare that financial support was received for the research, authorship, and/or publication of this article. Funding for this project was provided by the Jiangxi Provincial Education Department Science and Technology Research Project (Grant No. GJJ2203510) and the Jiangxi Provincial Health Commission Science and Technology Project (Grant No. 202211164).