Distinct BNC1 variants contribute to premature ovarian insufficiency through different molecular mechanisms

Premature ovarian insufficiency (POI) is a major cause of female infertility, and accumulating evidence indicates that genetic variants contribute to its pathogenesis. To further explore the genetic basis of POI, we performed a whole-exome sequencing among 121 women with POI and identified three rare heterozygous BNC1 gene variants, which include two missense variants, p.(Leu532Pro) and p.(Ser938Asn), and one truncating variant, p.(Arg15Profs*19). Basonuclin 1 (BNC1), a zinc finger transcription factor, has previously been associated with familial forms of POI, suggesting its important role in ovarian development and transcriptional regulation. Herein, we constructed wild-type (WT) and three mutant plasmids of BNC1 and performed RNA sequencing (RNA-seq) to characterize their functional consequences. Each BNC1 variant induced a distinct set of differentially expressed genes compared with the WT. Gene ontology enrichment analysis revealed that the p.(Leu532Pro) was associated with dysregulation of immune-related processes, p.(Ser938Asn) affected antiviral and innate immune responses, and truncating p.(Arg15Profs*19) altered reproductive and developmental pathways. Quantitative real-time polymerase chain reaction confirmed the differential expression of representative genes identified by RNA-seq. This study provides functional evidence that BNC1 variants contribute to POI through variant-specific molecular mechanisms, thereby providing further evidence to support BNC1 as a recurrent candidate gene in POI.