Identification of a novel nonsense mutation in α-galactosidase A that causes Fabry disease in a Chinese family

Abstract

Fabry disease, a lysosomal storage disease, is an uncommon X-linked recessive genetic disorder stemming from abnormalities in the alpha-galactosidase gene (GLA) that codes human alpha-Galactosidase A (α-Gal A). To date, over 800 GLA mutations have been found to cause Fabry disease (FD). Continued enhancement of the GLA mutation spectrum will contribute to a deeper recognition and underlying mechanisms of FD. In this study, a 27-year-old male proband exhibited a typical phenotype of Fabry disease. Subsequently, family screening for Fabry disease was conducted, and high-throughput sequencing was employed to identify the mutated gene. The three-level structure of the mutated protein was analyzed, and its subcellular localization and enzymatic activity were determined. Apoptosis was assessed in GLA mutant cell lines to confirm the functional effects. As a result, a new mutation, c.777_778del (p. Gly261Leufs*3), in the GLA gene was identified. The mutation caused a frameshift during translation and the premature appearance of a termination codon, which led to a partial deletion of the domain in C-terminal region and altered the protein’s tertiary structure. In vitro experiments revealed a significant reduction of the enzymatic activity in mutant cells. The expression was noticeably decreased at the mRNA and protein levels in mutant cell lines. Additionally, the subcellular localization of α-Gal A changed from a homogeneous distribution to punctate aggregation in the cytoplasm. GLA mutant cells exhibited significantly higher levels of apoptosis compared to wild-type cells.

Keywords:

• Fabry disease
• α-Gal A protein
• mutation
• enzymatic activity
• subcellular localization
• cell apoptosis

Authors’ contributions

Conceptualization: FX, HX, and YM; Data curation: YP and ZS; Formal analysis: YP, MP, and YW; Supervision: JX; Funding acquisition: YM; Writing – original draft: YP and YW.

Ethics statement

This study received approval from the Ethics Committee of Peking University Shenzhen Hospital, an affiliation of Peking University (2022-177). The research was conducted on the basis of the World Medical Association Declaration of Helsinki, and written informed consent was given in all subjects.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

Upon reasonable request, the relevant authors will make the data available.

Additional information

Funding

This study received funding from the National Natural Science Foundation of China (No. 82270784, 82070770) and the Guangdong Basic and Applied Basic Research Foundation (No. 2023A1515012276, 2024A1515012700).