Abnormal function of EPHA2/p.R957P mutant in congenital cataract
Authors:   Department of Ophthalmology, the First Affiliated Hospital of Fujian Medical University; Fujian Institute of Ophthalmology, Zhang Jing-Jin , Zhou Bi-Ting ,   Department of Ophthalmology, the First Affiliated Hospital of Fujian Medical University; Fujian Institute of Ophthalmology, Yang Ju-Hua ,   Department of Ophthalmology, the First Affiliated Hospital of Fujian Medical University; Fujian Institute of Ophthalmology; Chinese Academy of Medical Sciences &amp; Peking Union Medical College, Beijing 100000, China, Li Zhong ,   Chinese Academy of Medical Sciences &amp; Peking Union Medical College, Beijing 100000, China, Lin Shuang ,   Chinese Academy of Medical Sciences &amp; Peking Union Medical College, Beijing 100000, China, Chen Xiao-Le ,   Chinese Academy of Medical Sciences &amp; Peking Union Medical College, Beijing 100000, China, Zhang Nan-Wen ,   Chinese Academy of Medical Sciences &amp; Peking Union Medical College, Beijing 100000, China, Ye Qin ,   Department of Ophthalmology, the First Affiliated Hospital of Fujian Medical University; Fujian Institute of Ophthalmology, Ma Xu ,   Fujian Provincial Clinical Medical Research Center of Eye Diseases and Optometry, Fuzhou 350005, Fujian Province, China; Department of Ophthalmology, National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou 350005, Fujian Province, China, Zhu Yi-Hua ,   Department of Ophthalmology, the First Affiliated Hospital of Fujian Medical University; Fujian Institute of Ophthalmology
Published: May 2024

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 EPHA2/p.R957P mutant to demonstrate whether the mutant could cause lens opacity in vivo.
RESULTS: A novel missense and pathogenic variant c.2870G>C was identified in the sterile alpha motif (SAM) domain of EPHA2. 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 EPHA2/p.R957P mutant under differential interference contrast (DIC) optics.
CONCLUSION: Novel pathogenic c.2870G>C variant of EPHA2 in a Chinese congenital cataract family contributes to disease pathogenesis.