SLC26A4 C.317C > A Variant: Functional Analysis and Patient‐Derived Induced Pluripotent Stem Line Development

TL;DR

This study investigates how a specific SLC26A4 gene variant affects hearing loss by analyzing its impact on protein function and developing patient-derived stem cells for further research.

Contribution

The study introduces a novel in vitro system using patient-derived iPSCs to investigate the functional impact of the SLC26A4 c.317C > A variant.

Findings

The c.317C > A variant significantly reduces SLC26A4 mRNA and protein expression.

The variant causes Pendrin protein to accumulate as cytoplasmic aggregates.

Patient-derived iPSCs retained pluripotency, differentiation potential, and genetic integrity.

Abstract

SLC26A4 is the second most common cause of hereditary hearing loss worldwide. This gene predominantly harbors pathogenic variants, including splice, nonsense, and missense. Although missense variants are relatively common, their specific effects on protein function remain unclear. Consequently, there is an urgent need to establish an in vitro system to investigate how these variants impact SLC26A4 protein function.

Genetic testing was conducted to determine the specific types of underlying genetic variants in patients. Following this, we employed plasmid transfection to evaluate the effects of the variants on both protein expression levels and the protein's subcellular localization. Thereafter, we transformed peripheral blood mononuclear cells (PBMCs) from the proband into induced pluripotent stem cells (iPSCs) through Sendai virus‐mediated transduction.

Genetic testing revealed that the proband carried compound heterozygous variants: SLC26A4 c.919‐2A > G and c.317C > A. The c.317C > A variant markedly decreased the expression levels of SLC26A4 mRNA and its encoded protein. Additionally, it led to the protein's accumulation in the cytoplasm as aggregates. We successfully reprogrammed peripheral blood mononuclear cells from the proband into induced pluripotent stem cells (iPSCs) and verified that these iPSCs retained their pluripotency, differentiation potential, and genetic integrity.

These results provide important insights into the mechanisms by which SLC26A4 gene variants lead to hearing loss.

We identified compound heterozygous variants in the SLC26A4 gene, c.919‐2A > G and c.317C > A, and studied how the c.317C > A variant affects Pendrin expression and function. We successfully induced pluripotent stem cells (iPSCs) from peripheral blood mononuclear cells (PBMCs) of the proband.

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