Integrated Metabolomic and Transcriptomic Profiling Reveals a Distinct Pathological Aging State in Diminished Ovarian...

Women of advanced age exhibit significant heterogeneity in ovarian reserve, categorized as normal (NOR) or diminished (DOR). This study aims to distinguish physiological age-related decline from pathological accelerated aging in DOR, and to explore underlying molecular mechanisms for optimizing assisted reproductive strategies. In vitro fertilization-embryo transfer (IVF-ET) outcomes were retrospectively compared in advanced-age women (≥ 40 years) with NOR or DOR treated at our center (January 2022 - December 2024). Simultaneously, follicular fluid (FF) was collected from both groups (n = 20). After centrifugation, metabolomic analysis was performed on metabolites, and transcriptomic sequencing on isolated granulosa cells (GCs). Despite comparable fertilization and cleavage-stage embryo quality, the DOR group showed significantly lower rates of oocyte maturation, blastocyst formation, clinical pregnancy, and live birth (P < 0.05). Metabolomic analysis revealed 28 differential metabolites (DMs) in FF, primarily enriched in galactose metabolism. Transcriptomics of GCs identified 246 differentially expressed mRNAs (DEmRNAs), prominently enriched in immune-related pathways. Protein-protein interaction analysis highlighted five hub genes (CX3CR1, CD69, FCER1A, EOMES, SPRR2A). Integrated analysis of the top 50 DEmRNAs with the top 400 DEmRNA-DM correlation pairs identified five key genes-IGLC3, RNVU1-29, FAM110C, NPY2R, and KCNN4-bridging GC transcriptome and FF metabolome, with key pairs including RNVU1-29 with lysylhydroxyproline and a sterane derivative, and FAM110C with 16-hydroxyhexadecanoic acid. In conclusion, DOR in advanced age may represent a distinct pathological aging state characterized by a dysregulated follicular microenvironment potentially shaped by immune activation and metabolic reprogramming. The identified key gene-metabolite pairs offer candidate molecular links to compromised oocyte developmental competence.