Research Paper Volume 13, Issue 5 pp 7052—7066

Infertility network and hub genes for nonobstructive azoospermia utilizing integrative analysis

Baoquan Han1, *, , Zihui Yan2, *, , Shuai Yu1, *, , Wei Ge2, , Yaqi Li3, , Yan Wang1, , Bo Yang4, , Wei Shen1,2, , Hui Jiang5, , Zhongyi Sun1, ,

  • 1 Department of Urology, Peking University Shenzhen Hospital, Shenzhen Peking University, The Hong Kong University of Science and Technology Medical Center, Shenzhen 518036, China
  • 2 College of Life Sciences, Institute of Reproductive Sciences, Qingdao Agricultural University, Qingdao 266109, China
  • 3 Department of Urology, Zaozhuang Hospital of Zaozhuang Mining Group, Zaozhuang 277100, China
  • 4 Guangdong and Shenzhen Key Laboratory of Male Reproductive Medicine and Genetics, Institute of Urology, Peking University Shenzhen Hospital, Shenzhen PKU-HKUST Medical Center, Shenzhen 518036, China
  • 5 Department of Urology, Department of Andrology, Department of Human Sperm Bank, Peking University Third Hospital, Beijing 100191, China
* Equal contribution

Received: November 5, 2020       Accepted: December 29, 2020       Published: February 17, 2021
How to Cite

Copyright: © 2021 Han et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Non-obstructive azoospermia (NOA) is the most severe form of male infertility owing to the absence of sperm during ejaculation as a result of failed spermatogenesis. The molecular mechanisms of NOA have not been well studied. Here, we revealed the dysregulated differentially expressed genes in NOA and related signaling pathways or biological processes. Cluster features of biological processes include spermatogenesis, fertilization, cilium movement, penetration of zona pellucida, sperm chromatin condensation, and being significantly enriched metabolic pathways in proximal tubule bicarbonate reclamation, aldosterone synthesis and secretion, glycolysis and glycogenesis pathways in NOA using Gene Ontology analysis and pathway enrichment analysis. The NOA gene co-expression network was constructed by weighted gene co-expression network analysis to identify the hub genes (CHD5 and SPTBN2). In addition, we used another Gene Expression Omnibus dataset (GSE45887) to validate these hub genes. Furthermore, we used the Seurat package to classify testicular tissue cells from NOA patients and to characterize the differential expression of hub genes in different cell types from different adult males based on the scRNA-seq dataset (GSE106487). These results provide new insights into the pathogenesis of NOA. Of particular note, CHD5 and SPTBN2 may be potential biomarkers for the diagnosis and treatment of NOA.


NOA: nonobstructive azoospermia; TPM: Transcripts Per Kilobase Million; WGCNA: Weighted Gene Co-expression Network Analysis; scRNA-seq: Single-cell transcriptome sequencing; DEGs: Differentially Expressed Genes; GSEA: Gene Set Enrichment Analysis; PPI: Protein-Protein interaction; GEO: Gene Expression Omnibus; SRA: Sequence Read Archive; GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; STRING: Search Tool for the Retrieval of Interacting Genes/Proteins; OA: Obstructive Azoospermia.