A panel of extracellular vesicle long noncoding RNAs in seminal plasma for predicting testicular spermatozoa in nonobstructive azoospermia patients

Abstract

STUDY QUESTION Whether the testis-specific extracellular vesicle (EV) long noncoding RNAs (lncRNAs) in seminal plasma could be utilized to predict the presence of testicular spermatozoa in nonobstructive azoospermia (NOA) patients? SUMMARY ANSWER Our findings indicate that the panel based on seminal plasma EV lncRNAs was a sensitive and specific method in predicting the presence of testicular spermatozoa and may improve clinical decision-making of NOA. WHAT IS KNOWN ALREADY The adoption of sperm retrieval techniques, especially microdissection testicular sperm extraction (mTESE), in combination with ICSI has revolutionized treatment for NOA. However, there are no precise and noninvasive methods for predicting whether there are testicular spermatozoa in NOA patients before mTESE. STUDY DESIGN, SIZE, DURATION RNA sequencing was performed on seminal plasma EVs from 6 normozoospermic men who underwent IVF due to female factor and 5 idiopathic NOA patients who failed to obtain testicular spermatozoa by mTESE and were diagnosed as having Sertoli cell-only syndrome by postoperative pathology. A biomarker panel of lncRNAs was constructed and verified in 96 NOA patients who underwent mTESE. Decision-making process was established based on the panel in seminal plasma EVs from 45 normozoospermia samples, 43 oligozoospermia samples, 62 cryptozoospermia samples, 96 NOA samples. PARTICIPANTS/MATERIALS, SETTING, METHODS RNA sequencing was done to examine altered profiles of EV lncRNAs in seminal plasma. Furthermore, a panel consisting of EV lncRNAs was established and evaluated in training set and validation sets. MAIN RESULTS AND THE ROLE OF CHANCE A panel consisting of nine differentially expressed testis-specific lncRNAs, including LOC100505685, SPATA42, CCDC37-DT, GABRG3-AS1, LOC440934, LOC101929088 (XR_927561.2), LOC101929088 (XR_001745218.1), LINC00343 and LINC00301, was established in the training set and the AUC was 0.986. Furthermore, the AUC in the validation set was 0.960. Importantly, the panel had a unique advantage when compared with models based on serum hormones from the same group of NOA cases (AUC, 0.970 vs 0.723; 0.959 vs 0.687, respectively). According to the panel of lncRNAs, a decision-making process was established, that is when the score of an NOA case exceeds 0.532, sperm retrieval surgery may be recommended. LIMITATIONS, REASONS FOR CAUTION In the future, the sample size needs to be further expanded. Meanwhile, the regulatory functions and mechanism of lncRNAs in spermatogenesis also need to be elucidated. WIDER IMPLICATIONS OF THE FINDINGS When the score of our panel is below 0.532, subjecting the NOA patients to ineffective surgical interventions may not be recommended due to poor sperm retrieval rate. STUDY FUNDING/COMPETING INTEREST(S) This work was supported by the National Natural Science Foundation of China (81871110, 81971314 and 81971759); the Guangdong Special Support Plan-Science and Technology Innovation Youth Top Talents Project (2016TQ03R444); the Science and Technology Planning Project of Guangdong Province (2016B030230001 and 201707010394); the Key Scientific and Technological Program of Guangzhou City (201604020189); the Pearl River S&T Nova Program of Guangzhou (201806010089); the Transformation of Scientific and Technological Achievements Project of Sun Yat-sen University (80000-18843235) and the Youth Teacher Training Project of Sun Yat-sen University (17ykpy68 and 18ykpy09). There are no competing interests related to this study. TRIAL REGISTRATION NUMBER N/A.

Testicular germ cell–specific lncRNA, Teshl , is required for complete expression of Y chromosome genes and a normal offspring sex ratio

“…1D ). It was recently reported that LINC01921 is significantly decreased or differentially expressed in patients with nonobstructive azoospermia ( 49 ). Although further investigation is required to determine whether a decrease in the expression levels of LINC01921 is directly involved in nonobstructive azoospermia, this human lncRNA Teshl ortholog could be considered a suitable biomarker for diagnosis of human infertility.…”

Comparison of the Effects of Different Testicular Sperm Extraction Methods on the Embryonic Development of Azoospermic Men in Intracytoplasmic Sperm Injection (ICSI) Cycles: A Retrospective Cohort Study

“…Çayan et al reported a study that NOA men with a history of cryptorchidism have high sperm retrieval rates with mTESE [ 9 ]. However, some studies have shown that there are no precise and noninvasive methods for predicting whether there are testicular spermatozoa in NOA patients before mTESE [ 10 ]. Maglia et al found that the success rate of sperm retrieval in NOA patients was similar between the mTESE group and the conventional TESA group, and mTESE seemed to be more beneficial for patients over 35 years of age and with high FSH value [ 11 ].…”

Hormonal markers as noninvasive predictors of sperm retrieval in non-obstructive azoospermia

“…Firstly, the predictive value of these biomarkers has predominantly been observed in only one study with a small number of NOA men and consequently is inconclusive and questionable. In support of this, while miR-539-5p emerged as a good predictor of successful sperm recovery in one study [50], it exhibited no predictive value regarding TESE outcome in another study [51]. Hence, it seems reasonable to assume that further examining the predictive potential of the aforementioned molecular biomarkers can provoke similar controversy observed for hormonal markers.…”

“…Likewise, a seminal panel of exosomal long non-coding RNAs (lncRNAs) elicited from NOA patients displayed an outstanding predictive value in terms of sperm retrieval rate according to a recent report. Interestingly, the aforementioned lncRNA panel had remarkably higher predictive precision for the recovery of sperm from the testicular biopsies of NOA men in comparison with two different hormone-based models [51]. Moreover, there are numerous studies that evaluated seminal plasma proteome in azoospermic patients, although the majority of them have focused on the diagnosis and differentiation of azoospermia subclasses, i.e., OA and NOA, rather than the success of TESE outcome.…”