The Novel Hub Genes of Klinefelter Syndrome Affect Sperm Cell Differentiation: Analysis Based on RNA-Seq and scRNA-Seq Data

Objective: In this study, the Hub genes that are closely associated with spermatogenesis in klinefelter syndrome (KS) patients were identified, mainly through transcriptomic and single-cell RNA sequencing (scRNA-seq)data. Methods Transcriptome data were obtained from 3 adult 4 fetal KS patients and 8 normal individuals. ScRNA-seq data were obtained from 1 adult KS patient and 4 normal individuals. The differentially expressed genes (DEGs) were analysed using limma's method. The testicular tissue-specific protein-protein interaction (PPI) networks were analysed through the NetworkAnalyst website. The CytoHubba plugin was used to draw Veen Diagram. Integration of scRNA-seq data using the Seurat package. Developmental trajectories of sperm cells and hub genes were constructed by using the monocle3. Results The results of the GSE103905 and GSE103613 transcriptome datasets show a total of 700 DEGs. Gene Ontology (GO) analysis focused on memory, methylation, ATPase activity and mitochondrion. Kyoto Encyclopedia of Genes and Genomes (KEGG) focused on metabolic pathways, huntington's disease and nicotinamide metabolism. Four Hub genes (KIF2C, MRPS2, RPS15, TSFM) that have not been reported to be associated with KS were identified. Single-cell sequencing data showed that only KIF2C and RPS15 were expressed in sperm cells. hub genes' Pseudotime analysis showed a decreasing trend in RPS15 expression and an increasing trend in KIF2C expression throughout sperm development. Conclusion In this study, two RPS15 and KIF2C were identified that may be closely associated with spermatogenesis in KS patients, but there may be differences in the mechanisms by which they affect sperm development.