A novel hsa_circ_۰۰۷۰۹۶۳/miR-۲۲۳-۳p/CCDC۹۶–CCDC۱۱۲ 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_۰۰۷۰۹۶۳/miR-۲۲۳-۳p/CCDC۹۶–CCDC۱۱۲ as a potential biomarker for predicting microsurgical testicular sperm extraction (micro-TESE) outcomes in NOA individuals. Materials and Methods: In this case-control study, ۶۰ men with NOA undergoing micro-TESE and ۴۰ 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 CCDC۹۶ and CCDC۱۱۲ in NOA plasma, particularly in NOA⁻ individuals, while hsa-miR-۲۲۳-۳p was upregulated, with the highest expression in NOA⁻ cases. Conversely, hsa_circ_۰۰۷۰۹۶۳ 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_۰۰۷۰۹۶۳ (area under the receiver operating characteristic curve = ۰.۹۸۳) and hsa-miR-۲۲۳-۳p (area under the curve = ۰.۹۷۰) outperforming CCDC۹۶ and CCDC۱۱۲.   Conclusion: The hsa_circ_۰۰۷۰۹۶۳/miR-۲۲۳-۳p/CCDC۹۶–CCDC۱۱۲ 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.

کلیدواژه ها:

Azoospermia ، circRNA ، miRNA. ، آزواسپرمی ، circRNA ، miRNA.