Single-cell, spatial and bulk transcriptome data analysis revealed LINC00467-mediated Sertoli cell ferroptosis is a potential therapeutic target and biomarker for azoospermia.

Human spermatogenesis is an important physiological process related to programmed cell death. However, which type of programmed cell death playing a key role in normal and abnormal human spermatogenesis remains obscure. This study integrated single-cell, bulk RNA and spatial transcriptome data analysis and found that the ferroptosis signal plays a potential role in spermatogenesis and significantly elevate in testicular samples from humans with non-obstructive azoospermia (NOA) due to various factors. In addition, the increased levels of GPX4-dependent ferroptosis in Sertoli cells is a common feature of NOA samples and inhibiting ferroptosis of Sertoli cells in azoospermia patients can improve the functional status of Sertoli cells. Based on weighted correlation network analysis (WGCNA), correlation analysis, differential expression analysis and in vitro experiments, we confirmed that long non-coding RNA (lncRNA) LINC00467 is a key factor in regulating ferroptosis in Sertoli cells. Finally, quantitative real-time polymerase chain reaction (qRT-PCR) experiments showed that LINC00467 can be used to predict sperm retrieval outcomes in NOA patients. In summary, this study reveals and verifies that the level of ferroptosis apparently elevates in Sertoli cells is a novel mechanism for NOA patients, and proposes that ferroptosis-related gene LINC00467 is potential biomarker and therapeutic target for NOA.