Single-cell multi-omics analysis reveals dysfunctional Wnt signaling of spermatogonia in non-obstructive azoospermia

Frontiers in Endocrinology

Endocrinology

Xvdong Liu¹  Haibin Tang²  Liuxun Chen³  Chuan Liu³  Shengjie Zeng³  Hao Wu³ 

[1] Department of Cardiothoracic Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China;Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China;Department of Urology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China;

关键词: non-obstructive azoospermia;    ScRNA-seq;    scATAC-seq;    spatial transcriptomic data;    Wnt signaling pathway;    CCCTC-binding factor (CTCF);    androgen receptor (AR);    ARNTL;   

DOI  :  10.3389/fendo.2023.1138386

 received in 2023-01-05, accepted in 2023-05-17,  发布年份 2023

来源: Frontiers

【 摘 要 】

BackgroundNon-obstructive azoospermia (NOA) is the most severe type that leads to 1% of male infertility. Wnt signaling governs normal sperm maturation. However, the role of Wnt signaling in spermatogonia in NOA has incompletely been uncovered, and upstream molecules regulating Wnt signaling remain unclear.MethodsBulk RNA sequencing (RNA-seq) of NOA was used to identify the hub gene module in NOA utilizing weighted gene co-expression network analyses (WGCNAs). Single-cell RNA sequencing (scRNA-seq) of NOA was employed to explore dysfunctional signaling pathways in the specific cell type with gene sets of signaling pathways. Single-cell regulatory network inference and clustering (pySCENIC) for Python analysis was applied to speculate putative transcription factors in spermatogonia. Moreover, single-cell assay for transposase-accessible chromatin sequencing (scATAC-seq) determined the regulated genes of these transcription factors. Finally, spatial transcriptomic data were used to analyze cell type and Wnt signaling spatial distribution.ResultsThe Wnt signaling pathway was demonstrated to be enriched in the hub gene module of NOA by bulk RNA-seq. Then, scRNA-seq data revealed the downregulated activity and dysfunction of Wnt signaling of spermatogonia in NOA samples. Conjoint analyses of the pySCENIC algorithm and scATAC-seq data indicated that three transcription factors (CTCF, AR, and ARNTL) were related to the activities of Wnt signaling in NOA. Eventually, spatial expression localization of Wnt signaling was identified to be in accordance with the distribution patterns of spermatogonia, Sertoli cells, and Leydig cells.ConclusionIn conclusion, we identified that downregulated Wnt signaling of spermatogonia in NOA and three transcription factors (CTCF, AR, and ARNTL) may be involved in this dysfunctional Wnt signaling. These findings provide new mechanisms for NOA and new therapeutic targets for NOA patients.

【 授权许可】

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Copyright © 2023 Zeng, Chen, Liu, Tang, Wu and Liu