Transcriptomic survey of key reproductive and metabolic tissues in mouse models of polycystic ovary syndrome

Abstract

Excessive androgen production and obesity are key to polycystic ovary syndrome (PCOS) pathogenesis. Prenatal androgenized (PNA), peripubertal androgenized, and overexpression of nerve growth factor in theca cells (17NF) are commonly used PCOS-like mouse models and diet-induced maternal obesity model is often included for comparsion. To reveal the molecular features of these models, we have performed transcriptome survey of the hypothalamus, adipose tissue, ovary and metaphase II (MII) oocytes. The largest number of differentially expressed genes (DEGs) is found in the ovaries of 17NF and in the adipose tissues of peripubertal androgenized models. In contrast, hypothalamus is most affected in PNA and maternal obesity models suggesting fetal programming effects. The Ms4a6e gene, membrane-spanning 4-domains subfamily A member 6E, a DEG identified in the adipose tissue in all mouse models is also differently expressed in adipose tissue of women with PCOS, highlighting a conserved disease function. Our comprehensive transcriptomic profiling of key target tissues involved in PCOS pathology highlights the effects of developmental windows for androgen exposure and maternal obesity, and provides unique resource to investigate molecular mechanisms underlying PCOS pathogenesis.

Fig. 1. Summary of metabolic and reproductive phenotype and Illustration of three PCOS-like mouse models and a maternal high-fat high sugar (HFHS)-treated mouse model for comparative transcriptomic analyses of collected target tissues and MII oocytes.

a Summary of metabolic and reproductive phenotype of Prenatal androgenized (PNA), Peripubertal androgenized, 17NF, and maternal obesity. b PNA in which F1 female offspring was analyzed. c Peripubertal androgenized (subcutaneous DHT implant), and d 17NF model: theca-cell specific nerve growth factor (NGF) overexpressing transgenic mouse models. e F₁ female offspring of HFHS diet-induced maternal obese model. Different targeted tissues, namely hypothalamus, pituitary, ovaries, and subcutaneous adipose tissue are used for transcriptomic analysis by bulk mRNA sequencing. Metaphase II (MII) oocytes are used for single-cell RNA sequencing. N/A = not available.

Fig. 2. Common and distinct transcriptomic signature of the hypothalamus.

a Venn diagram of DEGs across all animal models (PNA model, n = 3 in control + vehicle, n = 2 in prenatal DHT; maternal obesity, n = 3 in control, n = 3 in maternal obesity; Peripubertal androgenized model, n = 4 in control + vehicle, n = 3 in Peripubertal DHT; 17NF, n = 3 in control, n = 3 in 17NF). b Comparison of enriched gene ontology terms of DEGs across all animal models. c–f Bar plot showing expression level of DEGs enriched in lipid metabolism, steroid hormone metabolism, and gonad development in PNA, Peripubertal androgenized, 17NF, and maternal obesity, respectively. g Illustration of DEGs involved in lipid and steroid metabolism pathways in hypothalamus (Created with BioRender.com). Data are presented as mean ± SD. n number of animals.

Fig. 3. Common and distinct transcriptomic signature of the ovary.

a Venn diagram of DEGs across all animal models (PNA model, n = 3 in control + vehicle, n = 4 in Prenatal DHT; peripubertal androgenized model, n = 4 in control + vehicle, n = 4 in Peripubertal DHT; 17NF, n = 3 in control, n = 4 in 17NF; maternal obesity, n = 3 in control, n = 3 in maternal obesity). b Comparison of enriched gene ontology term of DEGs across all animal models. c Heatmap of expression of DEGs enriched in Ovulation cycle and Steroid hormone metabolism in PNA, maternal obesity, Peripubertal androgenized, and 17NF respectively. d Overlapped DEG genes of ovary tissue with GWAS genes women with PCOS in PNA, maternal obesity, Peripubertal androgenized, and 17NF. e Module clustering tree diagram across animal models. Key module gene network involved in PCOS-like animal models. f Protein–Protein Interaction network of genes in black module (black module in e). PPI Data are retrieved from String database. n = number of animals.

Fig. 4. Common and distinct transcriptomic signature of MII oocytes.

a Venn diagram of DEGs of MII oocytes across all animal models (PNA model, n = 8/3 animals in control + vehicle, n = 10/2 animals in Prenatal DHT; maternal obesity, n = 8/3 animals in control, n = 15/3 animals in maternal obesity; peripubertal androgenized model, n = 56/8 animals in control + vehicle, n = 67/8 animals in peripubertal DHT; 17NF, n = 16/3 animals in control, n = 16/3 animals in 17NF). n = number of MII oocytes. b Comparison of enriched gene ontology term of DEGs in MII oocytes across all animal models. c Overlap DEG genes of MII oocytes with GWAS genes women with PCOS in PNA, maternal obesity, Peripubertal androgenized, and 17NF. d–g Significant signaling pathways were ranked based on their differences in overall information flow within the inferred networks between control and treatment group in PNA, maternal obesity, Peripubertal androgenized, and 17NF, respectively.

Fig. 5. Metabolic pathway analysis of MII oocytes.

a–d Boxplot showing the key metabolic pathways in PNA (n = 8/3 animals in control + vehicle, n = 10/2 animals in prenatal DHT), Maternal obesity (n = 8/3 animals in control, n = 15/3 animals in maternal obesity), Peripubertal androgenized (n = 56/8 animals in control + vehicle, n = 67/8 animals in Peripubertal DHT) and 17NF model (n = 16/3 animals in control, n = 16/3 animals in 17NF), respectively. Violin plot with an overlay of boxplots showing the DEGs expression in each metabolic pathway. For the boxplot within each violin plot, middle lines indicate median values, boxes range from the 25th to 75th percentiles. n number of MII oocytes.

Fig. 6. Common and distinct transcriptomic signature of adipose tissue.

a Venn diagram of DEGs across all animal models (PNA model, n = 3 in control + vehicle, n = 4 in Prenatal DHT; maternal obesity, n = 3 in control, n = 3 in maternal obesity; Peripubertal androgenized model, n = 4 in control + vehicle, n = 4 in Peripubertal DHT; 17NF, n = 3 in control, n = 3 in 17NF). b Comparison of enriched gene ontology term of DEGs in adipose tissue across all animal models. c Heatmap of expression of DEGs enriched in Response to steroid hormone, inflammatory response in PNA, maternal obesity, Peripubertal androgenized, and 17NF, respectively. d Scatter plot comparing alteration of gene expression between disease and controls in mouse models and patients. Yellow dots indicate the genes whose expression alterations in mouse models are in line with patients. Blue dots indicate the genes whose expression alterations in mouse models are opposite to patients. The spearman correlation coefficient in corresponding group is also shown. n = number of animals.

Fig. 7. Transcriptomic interaction among hypothalamus, ovary, adipose tissue, and MII oocytes in PCOS-like models.

a Transcriptomic crosstalk and unique gene signatures among hypothalamus, ovary, adipose tissue, and MII oocytes in PNA, maternal obesity, Peripubertal androgenized, and 17NF, respectively.