Integrated Transcriptome and Proteome Analyses Reveal Differentially Expressed Genes and Proteins in Granulosa Cells from Female Patients with Metabolic Syndrome-associated Infertility

Metabolic Syndrome (MS) is a cluster of conditions that significantly increase the risk of infertility in women. Granulosa cells are crucial for ovarian folliculogenesis and fertility. Understanding molecular alterations in these cells can provide insights into MS-associated infertility. This study aimed to investigate Differentially Expressed Genes (DEGs) and Proteins (DEPs) in granulosa cells from female patients with MS-associated infertility. Transcriptome and proteome analyses were integrated to compare granulosa cells from three MS patients with infertility to three control subjects. RNA sequencing and quantitative proteomics analyses were conducted, followed by differential expression analysis, Gene Set Enrichment Analysis (GSEA), and Protein-protein Interaction (PPI) network construction. Functional enrichment of overlapping DEGs and DEPs and potential drug-protein interactions were also explored. Hub genes identified by PPI were validated via quantitative Polymerase Chain Reaction (qPCR) and western blot assays. Principal Component Analysis (PCA) demonstrated a distinct separation between MS and control groups, indicating significant differences in gene and protein expression. A total of 1,046 upregulated and 23 downregulated DEGs, along with 222 upregulated and 412 downregulated DEPs, were identified in the MS group. GSEA highlighted enrichment in processes, like the cell cycle and immune response. Venn diagram revealed 71 overlapping DEGs and DEPs, mainly related to immune regulation. Key hub proteins and potential therapeutic candidates were identified, with hub genes upregulated at the mRNA level, but downregulated at the protein level in granulosa cells of MS patients. The integrative analyses revealed significant molecular alterations in granulosa cells from MS patients with infertility. Identified DEGs, DEPs, and hub proteins suggested potential therapeutic targets and pathways for addressing MS-associated infertility.