Abnormal function of /p.R957P mutant in congenital cataract.

AIM

To identify genetic defects in a Chinese family with congenital posterior polar cataracts and assess the pathogenicity.

METHODS

A four-generation Chinese family affected with autosomal dominant congenital cataract was recruited. Nineteen individuals took part in this study including 5 affected and 14 unaffected individuals. Sanger sequencing targeted hot-spot regions of 27 congenital cataract-causing genes for variant discovery. The pathogenicity of the variant was evaluated by the guidelines of American College of Medical Genetics and InterVar software. Confocal microscopy was applied to detect the subcellular localization of fluorescence-labeled ephrin type-A receptor 2 (EPHA2). Co-immunoprecipitation assay was implemented to estimate the interaction between EphA2 and other lens membrane proteins. The mRNA and protein expression were analyzed by reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting assay, respectively. The cell migration was analyzed by wound healing assay. Zebrafish model was generated by ectopic expression of human /p.R957P mutant to demonstrate whether the mutant could cause lens opacity .

RESULTS

A novel missense and pathogenic variant c.2870G>C was identified in the sterile alpha motif (SAM) domain of . Functional studies demonstrated the variant's impact: reduced EPHA2 protein expression, altered subcellular localization, and disrupted interactions with other lens membrane proteins. This mutant notably enhanced human lens epithelial cell migration, and induced a central cloudy region and roughness in zebrafish lenses with ectopic expression of human /p.R957P mutant under differential interference contrast (DIC) optics.

CONCLUSION

Novel pathogenic c.2870G>C variant of in a Chinese congenital cataract family contributes to disease pathogenesis.