Seminal plasma extracellular vesicles tRF-Val-AAC-010 can serve as a predictive factor of successful microdissection testicular sperm extraction in patients with non-obstructive azoospermia

Abstract

Background There is a lack of biomarkers for distinguishing non-obstructive azoospermia (NOA) patients with successful sperm retrieval (Sp+) from those with failed sperm retrieval (Sp-). This study aimed to determine the potential of extracellular vesicles tRNA-derived small RNA (tsRNA) as a novel non-invasive biomarker for successful sperm retrieval by microdissection testicular sperm extraction (mTESE). Methods The study included 18 patients with NOA with successful sperm retrieval (Sp+) and 23 patients with NOA with failed sperm retrieval (Sp-), 15 obstructive azoospermia (OA) patients, 5 idiopathic oligospermia (IO) patients, and 12 healthy people. Seminal plasma extracellular vesicles tsRNA levels were used in a two-stage case-control study (screened by tsRNA sequencing on Illumina NextSeq instrument and validated by qRT-PCR). The bioinformatic analysis was performed to determine the role of tsRNA in the pathogenesis of non-obstructive azoospermia. Results Two tsRNAs (tRF-Val-AAC-010: AUC = 0.96, specificity = 80%, sensitivity = 95%; tRF-Pro-AGG-003: AUC = 0.96, specificity = 87%, sensitivity = 95%) were found to have high predictive accuracy for distinguishing the origin of azoospermia. In addition, the extracellular vesicles tRF-Val-AAC-010 resulted in high predictive ability (AUC = 0.89, sensitivity = 72%, specificity = 91%, P < 0.0001) in predicting the presence of sperm in non-obstructive azoospermia undergoing mTESE. Finally, bioinformatic analysis revealed that tRF-Val-AAC-010 were involved in spermatogenesis. Conclusions This study identified that the extracellular vesicles tRF-Val-AAC-010 and tRF-Pro-AGG-003 are biomarkers for the diagnosis of non-obstructive azoospermia, and that tRF-Val-AAC-010 as a potential non-invasive biomarker for predicting the presence of sperm in non-obstructive azoospermia testicular tissue.

Seminal plasma biomarkers for predicting successful sperm retrieval in patients with nonobstructive azoospermia: a narrative review of human studies

“…The predictive model provided a high predictive accuracy (AUC = 0.927). China 2022 Han et al [ 31 ] 18 NOA men with micro-TESE(+) and 23 NOA men with micro-TESE(-). tsRNAs The extracellular vesicle tRF-Val-AAC-010 resulted in high predictive ability (AUC = 0.89, sensitivity = 72%, specificity = 91%, P < 0.0001) for predicting the presence of sperm in nonobstructive azoospermia undergoing micro-TESE.…”

“…The human SP contains approximately 700 extracellular RNAs that are involved in regulating vesicle-mediated transport, protein kinase inhibition, and cell response to zinc, which may reflect the spermatogenesis state in the testis [ 32 ]. To date, the biomarkers studied in studies of SP RNAs difference between NOA patients with positive and negative surgical sperm extraction include microRNAs [ 26 , 30 ], long noncoding RNAs [ 28 ], circRNAs [ 29 ], and tsRNAs [ 31 ].…”

Using Small Non-Coding RNAs in Extracellular Vesicles of Semen as Biomarkers of Male Reproductive System Health: Opportunities and Challenges

“…Some additional efforts have been made to identify sEV-contained sncRNAs to discriminate the presence or the absence of sperm in a testicular biopsy with a spermatogenic disorder with high diagnostic accuracy. Outcome prediction of TESE (testicular sperm extraction) [ 40 ] and micro_TESE (microdissection TESE) [ 62 , 63 ] is relevant before azoospermic individuals are referred for assisted reproduction treatment (ART) because they can only benefit from ART procedures if testicular sperm is present.…”

“…( B ) This attribute should be considered for a proper interpretation of the sncRNA expression analysis (a cluster analysis is shown as an example) and the selection of semen EV sncRNA as biomarkers for urogenital disease. ( C ) Male reproductive seminal sEV sncRNA of interest are summarized in the tables [ 39 , 58 , 60 , 61 , 62 , 63 , 66 , 67 , 68 ].…”

Messenger roles of extracellular vesicles during fertilization of gametes, development and implantation: Recent advances

“…Furthermore, SP-EVs have some diagnostic value. SP-EV’s contents can reflect male infertility sperm status; for example, two tsRNA TRF-Val-AAC-010 and TRF-pro-AGG-003 in seminal plasma can be used to diagnose non-obstructive azoospermia ( Han et al, 2022 ). SP-EVs, as messengers communicating with sperm in seminal plasma, can indicate the status of the sperm.…”