An Oxidative Stress-Related Gene Signature in Granulosa Cells Is Associated with Ovarian Aging.

Ovarian aging is associated with a decrease in fecundity. Increased oxidative stress of granulosa cells (GCs) is an important contributor. We thus asked whether there is an oxidative stress-related gene signature in GCs associated with ovarian aging. Public nonhuman primate (NHP) single-cell transcriptome was processed to identify GC cluster. Then, a GC signature for ovarian aging was established based on six oxidative stress-related differentially expressed genes ( MAPK1 , STK24 , AREG , ATG7 , ANXA1 , and PON2 ). Receiver operating characteristic (ROC) analysis confirmed good discriminating capacity in both NHP single-cell and human bulk transcriptome datasets. Gene expression levels were investigated using qPCR in the human ovarian granulosa-like tumor cell line (KGN) and mouse GCs. In an oxidative stress model, KGN cells were treated with menadione (7.5  μ M, 24 h) to induce oxidative stress, after which upregulation of MAPK1 , STK24 , ATG7 , ANXA1 , and PON2 and downregulation of AREG were observed ( p < 0.05). In an aging model, KGN cells were continuously cultured for 3 months, leading to increased expressions of all genes ( p < 0.05). In GCs of reproductively aged (8-month-old) Kunming mice, upregulated expression of Mapk1 , Stk24 , Atg7 , and Pon2 and downregulated expression of Anxa1 and Areg were observed ( p < 0.01). We therefore here identify a six-gene GC signature associated with oxidative stress and ovarian aging.