The Novel Key Genes of Non-obstructive Azoospermia Affect Spermatogenesis: Transcriptomic Analysis Based on RNA-Seq and scRNA-Seq Data

Abstract

Non-obstructive azoospermia (NOA) is one of the most important causes of male infertility. It is mainly characterized by the absence of sperm in semen repeatedly or the number of sperm is small and not fully developed. At present, its pathogenesis remains largely unknown. The goal of this study is to identify hub genes that might affect biomarkers related to spermatogenesis. Using the clinically significant transcriptome and single-cell sequencing data sets on the Gene Expression Omnibus (GEO) database, we identified candidate hub genes related to spermatogenesis. Based on them, we performed Gene Ontology (GO) functional enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment pathway analyses, protein-protein interaction (PPI) network analysis, principal component analysis (PCA), cell cluster analysis, and pseudo-chronological analysis. We identified a total of 430 differentially expressed genes, of which three have not been reported related to spermatogenesis (C22orf23, TSACC, and TTC25), and the expression of these three hub genes was different in each type of sperm cells. The results of the pseudo-chronological analysis of the three hub genes indicated that TTC25 was in a low expression state during the whole process of sperm development, while the expression of C22orf23 had two fluctuations in the differentiating spermatogonia and late primary spermatocyte stages, and TSACC showed an upward trend from the spermatogonial stem cell stage to the spermatogenesis stage. Our research found that the three hub genes were different in the trajectory of sperm development, indicating that they might play important roles in different sperm cells. This result is of great significance for revealing the pathogenic mechanism of NOA and further research.

Influence of the Postmortem/Storage Time of Human Corneas on the Properties of Cultured Limbal Epithelial Cells

“…As shown on Figure 1 D, the expression of all the 50 genes identified was dramatically reduced in the LPM/ST hLECs group. Amongst them, several are known to encode proteins that regulate particularly important epithelial cell functions such as apoptosis ( ST18 , CISD3 ) [ 39 , 40 ], differentiation ( TBR1 , BARX1 , NKX1-2 , FOXB1 , APOL1 , PROP1 , TSACC ) [ 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 ], proliferation ( CEBPD , MNX1-AS1 , GPR75 , FZD9 , GAS2L3 , PFKL , TPT1-AS1 , CYP17A1 , MIR22HG ) [ 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 ], and migration ( ARHGAP9 , ITGAM , EMILIN1 , ZNF554 , SOX8 ) [ 59 , 60 , 61 , 62 , 63 ]. Moreover, the involvement of some of these genes in the cellular functions listed above was validated further by our IPA analyses.…”

KIF2C affects sperm cell differentiation in patients with Klinefelter syndrome, as revealed by RNA‐Seq and scRNA‐Seq data

“…Klinefelter syndrome is an important cause of non‐obstructive azoospermia (NOA) and oligospermia, which is accompanied by impaired spermatogenesis, with 90% of patients presenting with azoospermia and 10% with severe oligospermia [ 19 , 20 ]. Furthermore, the effects of KS mosaicism may not be limited to germ cells；Ivan scholars [ 21 ] had found that KSM may be a factor in the pathogenic cascade in psychiatric and neurodegenerative disorders, particularly in cases where two or more X chromosomes are overrepresented.…”

Construction and external validation of a 5-gene random forest model to diagnose non-obstructive azoospermia based on the single-cell RNA sequencing of testicular tissue

“…In addition, from the perspective of pathogenesis, although many theories have been proposed to explain the pathogenesis of NOA, our understandings of the biological processes associated with NOA remains insufficient. Nowadays, the widespread applications of gene sequencing, especially scRNA-seq, have deepened the knowledge of NOA [ 25 , 26 ]. For instance, Wang et al had disclosed the unique role of autophagy homeostasis in the spermatogenesis of NOA cases through scRNA-seq analysis [ 27 ].…”