Abstract
PURPOSE: To identify novel pathogenic genes and variants responsible for oocyte degeneration and death. METHODS: Whole-exome sequencing was conducted in 78 individuals with primary infertility characterized with oocyte degeneration and death, followed by Sanger sequencing of candidate variants. Pathogenicity of identified COX15 variants was characterized through morphological assessment, AlphaFold3-based structural modeling and functional validation including Western blotting and immunofluorescence. RESULTS: We identified a compound-heterozygous COX15 variants and a homozygous COX15 variant in two affected individuals with oocyte degeneration. The variants c.C664T [p.R222C] was a previously reported recurrent pathogenic variant. The clinical records showed that majority of oocytes retrieved from affected individuals were immature and degenerated. Structural modeling assay showed that all the COX15 variants affected the binding pocket with heme o. The compound-heterozygous COX15 variants c.C905T [p.P302L] and c.C664T [p.R222C] in family 1 significantly decreased COX15 protein level in HeLa cells. While the homozygous variant c.C647T [p.P216L] decreased the COX1 protein level in oocytes from family 2 (II-1), implying the disruption of respiratory complex IV (COXIV) assembly. CONCLUSIONS: Our study identified two novel variants in COX15, and functional analysis confirmed the pathogenicity of the variants. Our findings expand the mutational spectrum of COX15 and making it a potential genetic diagnostic marker for those suffering from female infertility causing by oocyte degeneration.