Introduction: DNA damage may affect homeostasis on spermatogonial stem cells(SSCs), while the details relationship with male infertility still remains unclear. Therefore, it is important that further research into the mechanisms related to DNA damage and genomic stability on spermatogonial stem cells.

Methods: scRNA-seq datasets from Idiopathic non-obstructive azoospermia (NOA) and normal testes were collected and testicular cells were further annotated via UMAP. Based annotation on the sequencing data, WGCNA analysis on the deferentially expressed genes was conducted , LASSO regression and the MNC calculation algorithm in Cytoscape were carried out to find genes associated with DNA damage repair.

Results: It was found that SSCs were mainly concentrated in normal samples, and the differences in subcluster pathways reflected the heterogeneity of NOA. While the characteristics of the interaction between Leydig cells and other cells were clarified, and the importance of the PTN signaling pathway in SSCs development was discovered, which participates in SSCs development through SDC2. Combining the marker genes of SSCs and DNA damage-related genes in single-cell analysis, a PPI network was constructed. Through LASSO regression and the MNC calculation algorithm in Cytoscape, ATRX, DOT1L, and RUVBL2 were finally identified as key diagnostic genes.

Conclusion: Our results revealed predictable mechanisms of testicular micro-environment and DNA damage in the regulation of human SSCs and propose potential therapeutic targets for male infertility. Subsequently, further research to confirm the predicted potential mechanisms, pathways, and therapeutic targets should be conducted.

S. Karger AG, Basel.