Study on DNA Damage Gene in Spermatogonial Stem Cells from Idiopathic Nonobstructive Azoospermia: A Bioinformatics Investigation Based on scRNA-seq Data.

INTRODUCTION

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

METHODS

scRNA-seq datasets from idiopathic nonobstructive azoospermia (NOA) and normal testes were collected and testicular cells were further annotated via UMAP. Based on annotation of the sequencing data, WGCNA analysis on the differentially 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, 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.