Comprehensive Analysis of the Association Between Human Non-obstructive Azoospermia and Plasticisers via Single-Cell and Traditional RNA Sequencing Methods

Abstract

AbstractA father’s lifetime experience is a major risk factor for a range of diseases in an individual. The influences of exposure can also be transmitted to offspring. Previous studies demonstrated that plasticisers can damage the male offspring reproductive system, but the link between mammalian research and human non-obstructive azoospermia remains underexplored. Here, we analysed reproduction-related genes from four publicly available single-cell RNA-Seq datasets and RNA-Seq datasets on GEO databases to investigate the correlation between human non-obstructive azoospermia and exposure to plasticisers during pregnancy. The R software was used in statistical analyses. A total of 9 co-upregulated genes and 1 co-downregulated gene were found. The Gene Ontology enrichment analyses were performed using the “clusterProfiler” package. Single-cell analyses were conducted to visualise the gene expression pattern in testis cell subgroups. Immunohistochemical images were used to evaluate the level of protein in testicular organs from The Human Protein Atlas. GSVA results provided further functional annotations. Three signature genes, i.e. COL1A1, CYP17A1 and KIF11, might serve as promising diagnostic biomarkers in non-obstructive azoospermia caused by plasticisers. Our results provided a potential new protocol to evaluate the feasibility of life or clinical intervention in patients with non-obstructive azoospermia. We believe that these observations will improve our understanding of the potential mechanisms of plasticiser contributions to human non-obstructive azoospermia and help identify potential targets for immunotherapy in the future.