MicroRNA expression profiles in the seminal plasma of nonobstructive azoospermia patients with different histopathologic patterns

Abstract

Objective: To investigate microRNA (miRNA) expression profiles in the seminal plasma of nonobstructive azoospermia (NOA) patients with different histopathologic patterns and evaluate potential noninvasive diagnostic biomarkers of NOA. Design: Sequencing and validation using quantitative reverse transcription polymerase chain reaction (qRT-PCR). Setting: Reproductive center and research institute. Patient(s): Thirteen patients with NOA (7 Sertoli cell-only syndrome [SCOS] and 6 hypospermatogenesis to spermatogenesis arrest [SA]) and 7 normal fertile controls for sequencing, six samples per group for validation; 54 patients with NOA (27 SCOS and 27 SA) and 19 normal fertile controls for large-sample qRT-PCR analysis. Intervention(s): None. Main Outcome Measure(s): MicroRNA expression profiles in the seminal plasma of patients with NOA with different histopathologic patterns were assessed using high-throughput sequencing and validated using qRT-PCR. Result(s): There were 78 overexpressed and 132 underexpressed miRNAs in patients with SCOS and 32 up-regulated and 90 downregulated miRNAs in patients with SA compared with fertile men with normozoospermia. Two down-regulated and one upregulated miRNA were validated using qRT-PCR, which indicated that the qRT-PCR and sequencing results were basically consistent. Hsa-miR-34c-5p expression was significantly lower in the seminal plasma of patients with NOA than normal fertile controls. The area under the receiver operating characteristic curve(AUC) for hsa-miR-34c-5p was 0.979 and 0.987 in the seminal plasma of patients with SA and patients with SCOS, respectively, compared with normal fertile controls. The AUC was 0.799 for hsa-miR-34c-5p in the seminal plasma between patients with SA and patients with SCOS. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis of differentially expressed miRNA target genes revealed that the Notch signaling pathway was one of the most abundant signaling pathways. The expression of Hes5, an effector of the Notch signaling pathway, was significantly higher in the seminal plasma of patients with NOA than normal fertile controls. Conclusion(s): MicroRNA expression profiles in seminal plasma were altered in patients with NOA compared with normal fertile controls. The profiles differed in patients with NOA with different pathologic patterns. We speculate that miR-34c-5p in seminal plasma could be a potential noninvasive biomarker to diagnose patients with NOA and distinguish different pathologic types of NOA. The Notch signaling pathway may be involved in the pathogenesis of NOA.

Are seminal plasma microRNAs the next biomarkers for nonobstructive azoospermia?

“…In the current study, Zhang et al (4) show decreased expression of seminal plasma miR-34c-5p in men with NOA compared with that in fertile controls using high-throughput sequencing. The relative expression of miR-34c-5p is significantly lower in patients with Sertoli-cell-only syndrome than in men with spermatogenesis arrest.…”

Using microRNAs as molecular biomarkers for the evaluation of male infertility

“…However, they realized that this alteration in the expression of miR‐34c‐5p was also lower in SCOS cases than in SA cases. These findings suggest that miR‐34c‐5p as a non‐invasive biomarker can distinguish different patterns in non‐obstructive azoospermia (Zhang, Zhang, et al, 2021). Another recent paper has reported that the expression levels of miR‐122‐5p, miR‐34c‐5p, miR‐509‐3‐5p, miR‐449a and miR‐202‐5p decreased in azoospermic patients compared with normal men.…”

“…Non‐obstructive azoospermia mainly consists of two histopathologic patterns: Sertoli cell‐only syndrome (SCOS) and spermatogenesis arrest (SA) defect. In a related study, a total of 20 different seminal plasma samples were isolated and purified from seminal plasma by the miRNeasy Micro Kit (Qiagen), and a significant down‐regulation of miR‐34c‐5p was found in seminal plasma samples of patients with non‐obstructive azoospermia compared with normal fertile controls (Zhang, Zhang, et al, 2021). However, they realized that this alteration in the expression of miR‐34c‐5p was also lower in SCOS cases than in SA cases.…”

Small Noncoding RNAs in Reproduction and Infertility

“…Seminal plasma, which is partly derived from epididymal fluid, generates sncRNA profiles that implicate seminal plasma quality in and male fertility. miRNA profiles in seminal plasma are correlated with specific spermatogenic defects and, more broadly, in male infertility [ 217 , 218 , 219 ]. Effects of environmental exposures on spermatogenesis, such as heat stress, may also be reflected by the miRNA profile of seminal plasma [ 220 ].…”