Novel Mutations Identified in the Chinese Han Population with Keratoconus by Next-Generation Sequencing

Binbin Chen; Xiaoning Yu; Xin Zhang; Hao Yang; Yilei Cui; Xingchao Shentu

doi:10.1155/2022/9991910

Novel Mutations Identified in the Chinese Han Population with Keratoconus by Next-Generation Sequencing

J Ophthalmol. 2022 Feb 10:2022:9991910. doi: 10.1155/2022/9991910. eCollection 2022.

Authors

Binbin Chen¹, Xiaoning Yu¹, Xin Zhang¹, Hao Yang¹, Yilei Cui¹, Xingchao Shentu¹

Affiliation

¹ Eye Center of the Second Affiliated Hospital, School of Medicine, Zhejiang University, Zhejiang Provincial Key Lab of Ophthalmology, Hangzhou, Zhejiang Province, China.

Abstract

Aim: To identify novel mutations in keratoconus (KC) susceptibility genes in the Chinese Han population.

Methods: A total of fifty-two patients with primary KC were recruited. Blood samples were collected, and genomic DNA was isolated from peripheral blood leukocytes. The entire coding region, intron-exon junctions, and promoter regions of sixteen known KC susceptibility genes were screened with next-generation sequencing technology. All identified variants were further confirmed using the Sanger sequencing technology. The Sorting Intolerant from Tolerant (SIFT), MutationTaster, and PolyPhen 2 programs were used to predict the effect of amino acid substitution on protein.

Results: After removing twelve known SNPs (single nucleotide polymorphisms) and three variants predicted to be harmless, nine novel mutations were identified in eight of the fifty-two patients, including c.455C > T:p.P152L in FNDC3B; c.3636_3637del:p.R1212fs in COL4A4; c.5015G > T:p.R1672L, c.3798dupA:p.P1267fs, and c.28G > A:p.A10T in MPDZ; c.1940C > T:p.P647L in DOCK9; c.127_128insGGC:p.Q43delinsRQ in POLG; c.3019G > A:p.V1007I in IPO5; and c.624 + 7- > A in TGFBI. All nine mutations in the patients with KC were heterozygote.

Conclusion: This study enlarged the gene profile of KC and should be further confirmed by well-powered, genome-wide association studies (GWAS) of Han Chinese patients.