PROTEOMIC ANALYSIS OF PLASMA EXOSOMES FROM PCOS PATIENTS WITH AND WITHOUT HYPERANDROGENEMIA

Hongfeng Wang^{1, 2, 3, #}, shuiwang Hu^{4, #}, Cuilian Zhang^{2, 3}, Baoxia Gu^{2, 3}, Yan Zhao^{2, 3}, Qian Wang^{2, 3}, Yingpu Sun^{1, *}

¹Reproductive Medicine Center, the First Affiliated Hospital of Zhengzhou University, No.1 East Jianshe Rd, Zhengzhou 450052, China - ²Reproductive Medicine Center, the People’s Hospital of Zhengzhou University, No.7 Weiwu Rd, Zhengzhou 450003, China - ³Reproductive Medicine Center, Henan Provincial People’s Hospital, No.7 Weiwu Rd, Zhengzhou 450003, China - ⁴Department of Pathophysiology, Southern Medical University, No.1023-1063 South Shatai Rd, Guangzhou 510515, China

Purpose: To better understand the mechanisms underlying PCOS with hyperandrogenemia (PCOS-HA) and PCOS without hyperandrogenemia (PCOS-NHA) and to identify new potential biomarkers for accurate diagnosis. 

Methods: High-resolution label-free mass spectrometry was utilized first to study the differential proteome of plasma-derived exosomes from participants in a healthy Control study, PCOS-HA, and PCOS-NHA. And then the differentially expressed proteins (DEPs) were carried out by a series of bioinformatics analyses. Finally, two DEPs were verified by ELISA analysis. 

Results: Fifty-nine DEPs were identified between PCOS-HA and PCOS-NHA, among which 11 specific proteins were screened with Wayne’s analysis. The heatmap of cluster analysis indicated that the expression profiles of DEPs could be divided primarily into 10 clusters. Protein-protein interaction network analysis revealed that most of the DEPs may have direct or indirect interactions. GO clustering enrichment analysis showed that the DEPs were mainly enriched in the immune system according to biological processes, participated in catalytic activity regarding molecular functions, and located in an extracellular region from a cellular components perspective. KEGG pathways analysis indicated that DEPs were mainly enriched in PI3K-Akt cell-signaling pathway. Furthermore, the expressional levels of AGP1 and ZNF28 in plasma exosomes were validated by ELISA analyses, which were consistent with the proteomic results. 

Conclusions: This study has successfully screened DEPs of plasma exosomes between PCOS-HA and PCOS-NHA, and AGP1 and ZNF28 showed potential biomarkers for differential diagnosis of PCOS.