Characterization of extracellular vesicles in human seminal plasma: a multiomic analysis

Extracellular Vesicles (EVs) derived from seminal plasma have achieved considerable attention due to their potential role in male reproductive physiology and pathology. In this study, we employed a comprehensive proteomic and transcriptomic approach to investigate the molecular signatures of EVs isolated from human seminal plasma. EVs from Normozoospermic (NORMO,15), OligoAsthenoTeratozoospermic (OAT,12) and Azoospermic (AZO,12) subjects, were isolated by an in house modified polymer precipitation-based protocol and characterized for their size and morphology by NTA and TEM. Full proteomic analysis, by gel-free and gel-based approaches, revealed distinct protein profiles in each group, highlighting potential molecules and pathways implicated in sperm function and fertility. Furthermore, transcriptomic analysis confirmed the trend of general down-regulation of AZO and OAT groups compared to NORMO offering insights into the regulatory mechanisms underlying sperm development and function. Bioinformatic tools were applied for functional Omics analysis; integration of proteomic and transcriptomic data provided a comprehensive understanding of the cargo content and regulatory networks. This study contributes to elucidating the key role of EVs in the paracrine communication regulating spermatogenesis. The full understanding of these pathways not only suggests potential mechanisms regulating male fertility but also offers new insight for the development of diagnostic tools targeting male reproductive disorders.