A novel hsa_circ_0070963/miR-223-3p/ CCDC96-CCDC112 regulatory axis as a non-invasive biomarker for predicting sperm retrieval outcomes in non-obstructive azoospermia: A case-control study.

BACKGROUND

Non-obstructive azoospermia (NOA) is a severe cause of male infertility. Testicular sperm extraction has limited success, largely due to the absence of reliable, non-invasive biomarkers to predict outcomes. Emerging evidence highlights the role of non-coding RNAs in spermatogenesis, providing promising targets for biomarker development.

OBJECTIVE

This study investigated the competing endogenous RNA regulatory axis hsa_circ_0070963/miR-223-3p/CCDC96-CCDC112 as a potential biomarker for predicting microsurgical testicular sperm extraction (micro-TESE) outcomes in NOA individuals.

MATERIALS AND

METHODS

In this case-control study, 60 men with NOA undergoing micro-TESE and 40 fertile controls were recruited. Testicular histology categorized individuals into Sertoli cell-only syndrome, hypospermatogenesis, or maturation arrest. Individuals were further stratified into NOA⁺ (successful retrieval) and NOA⁻ (failed retrieval). Plasma RNA was extracted, and expression levels of candidate genes were measured by quantitative reverse transcription polymerase chain reaction.

RESULTS

Findings demonstrated significant downregulation of CCDC96 and CCDC112 in NOA plasma, particularly in NOA⁻ individuals, while hsa-miR-223-3p was upregulated, with the highest expression in NOA⁻ cases. Conversely, hsa_circ_0070963 was markedly reduced in NOA, especially in NOA⁻ individuals. Dysregulation was more pronounced in hypospermatogenesis and maturation arrest compared to Sertoli cell-only syndrome. Receiver operating characteristic analysis revealed strong predictive accuracy, with hsa_circ_0070963 (area under the receiver operating characteristic curve = 0.983) and hsa-miR-223-3p (area under the curve = 0.970) outperforming CCDC96 and CCDC112.

CONCLUSION

hsa_circ_0070963/miR-223-3p/CCDC96-CCDC112 axis represents a novel competing endogenous RNAs regulatory network linked to impaired spermatogenesis. Its expression profile provides mechanistic insights and serves as a robust, non-invasive biomarker for predicting micro-TESE outcomes, supporting improved stratification and individualized treatment strategies in male infertility.