Abstract: PURPOSE: To identify the potentially pathogenic mutations in patients with ectodermal dysplasia (ED) and to investigate the pathogenicity of mutations by functional studies. Methods: Eight Chinese ED patients were included in this study. Peripheral venous blood was taken from the patients and DNA was extracted. Whole-exome sequencing (WES) was performed using DNA samples. After quality control of the sequencing data, the potentially pathogenic mutations were screened. The pathogenicity of the mutations was predicted in silico. Immunofluorescence study and dual luciferase assays were performed to investigate the pathogenicity of the mutations. Results: The effective rates of all sequencing samples were above 97.5% and the error rates were less than 0.03%. The proportions of Q20 were more than 97.0%. The average sequencing depths of the target region were more than 90×. The sequencing data were acceptable for further analysis. After data screening, three missense mutations of EDA were detected, including c.959A>G, c.1073A>G and c.1001G>A. The allele frequency was low in population database for all three mutations and in silico analysis indicated all three mutations were disease-causing. Immunofluorescence analysis showed that p65 protein nuclear translocation was compromised by EDA mutations, dual luciferase assays also showed that the activation of NF-κB pathway was decreased by EDA mutations. Conclusions: This study identified EDA mutations in Chinese ED patients and further verified the pathogenicity of the mutations by functional studies, contributing to the understanding of the pathogenesis of ED.

Key words: Ectodermal dysplasia, Whole-exome sequencing, NF-κB signaling pathway;, Functional studies