The roles of NF-κB-activated theca cell subtypes in subclinical premature ovarian insufficiency

Early-onset ovarian insufficiency affects 1%-5% of women under the age of 40 years, but its early pathogenesis remains unclear. This exploratory study employed a small sample design to investigate cellular and molecular alterations in the follicular microenvironment of patients with early-onset ovarian insufficiency, with a particular focus on inflammatory pathways. Through single-cell and bulk RNA sequencing analysis of granulosa cells, we observed elevated expression patterns of nuclear factor kappa B (NF-κB) signaling pathway activation in patient samples. Single-cell sequencing (n = 1 per group) revealed 11 cell subtypes, among which three inflammatory subpopulations exhibited elevated NF-κB activity, particularly NF-κB-activated follicular membrane/stromal cells, which dominated in patient samples. Computational deconvolution analysis based on population RNA sequencing data (n = 4 per group) supported the increased proportion of NF-κB-activated cell subpopulations in patient samples, with high cross-platform consistency (r = 0.86, p < 0.001). Trajectory analysis indicated that after removing NF-κB-activated follicular membrane/stromal cells, the developmental pathways of granulosa cell subpopulations in the patient group converged with those in the control group. Although the sample size was limited (single-cell sequencing, n = 1 per group), computational validation methods supported these preliminary findings. This study suggests that the NF-κB signaling pathway may be activated in the follicular microenvironment of early ovarian insufficiency, where inflammatory cells may influence granulosa cell differentiation through specific molecular interactions. This provides preliminary insights into the disease pathogenesis and suggests potential research directions for early diagnosis and intervention strategies, which require further validation in larger sample-size studies.