Exosomal microRNAs in seminal plasma are markers of the origin of azoospermia and can predict the presence of sperm in testicular tissue

Abstract BackgroundThere is lack of accurate and non-invasive preoperative evaluation for improving microdissection testicular sperm success rate. tRNA-derived small RNA (tsRNAs) perform a variety of physiological functions and are related to many physiological and pathological processes. This study sought to identify the potential of exosomal tsRNA as a novel non-invasive diagnostic biomarker for non-obstructive azoospermia (NOA) with spermatogenic failure.MethodsSeminal plasma exosome tsRNA levels were used in a two-stage (screened by tsRNA sequencing on Illumina NextSeq instrument and validated by qRT-PCR) case-control designed study. The expression levels of the selected tsRNAs were further examined in the testicular tissues of NOA patients and obstructive azoospermia (OA) patients, and their underlying role in the pathogenesis of non-obstructive azoospermia was performed by bioinformatic analysis.ResultsIn this study, 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 a good predictive accuracy which also showed differential expression in testicular tissue between NOA patients and OA patients. We also found that the combinations of tRF-Val-AAC-010 and tRF-pro -AGG-003 have a better discriminating ability between NOA patients and OA patients than single biomarkers. Finally, the bioinformatic analysis showed that the two selected tsRNAs were involved in spermatogenesis.ConclusionThis study first evaluated tsRNAs as potential biomarkers for NOA diagnosis and identified the exosomal tRF-Val-AAC-010 and tRF-pro-AGG-003 in seminal plasma valuable as biomarkers for NOA diagnosis.

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Lipidomic profile of seminal plasma in non-obstructive azoospermia with sperm maturation arrest

Herald Urology ◽

10.21886/2308-6424-2021-9-4-30-39 ◽

2021 ◽

Vol 9 (4) ◽

pp. 30-39

Author(s):

S. I. Gamidov ◽

T. V. Shatylko ◽

A. Kh. Tambiev ◽

A. O. Tokareva ◽

V. V. Chagovets ◽

...

Keyword(s):

Lipid Profile ◽

Seminal Plasma ◽

Married Couple ◽

Testicular Biopsy ◽

Testicular Tissue ◽

Sperm Maturation ◽

Negative Ion ◽

Obstructive Azoospermia ◽

Maturation Arrest ◽

Negative Ion Mode

Introduction. The difference between obstructive and non-obstructive azoospermia with sperm maturation arrest is important for the choice of treatment tactics and adequate counseling of a married couple.Purpose of the study. The study aimed to assess the semen lipid profile in patients with sperm maturation arrest. Materials and methods. Samples of seminal plasma for lipid composition of 24 men with normozoospermia and 64 men with azoospermia were studied. Patients with azoospermia underwent microdissection testicular biopsy followed by the detection of testicular tissue pathology. Lipid extracts were analyzed by liquid chromatography with mass spectrometry. Lipid data were compared with the results of pathomorphological studies.Results. Comparison of two groups revealed a statistically significant concentration differences for 22 lipids detected in positive-ion mode and 11 lipids detected in negative-ion mode. Those lipids mainly belong to the classes hexosylceramides, sphingomyelins and phosphatidylcholines — simple ethers and oxidized lipids. In multivariate analysis, the following lipids were found to be statistically significant predictors of sperm maturation arrest: PC 16: 0_22: 6 lipid (β-coefficient: -0.73; 95% confidence interval (95% CI): -1.42 to -0.27; odds ratio (OR): 0.48; OR CI: 0.24 to 0.76; Wald's test: -2.58; p = 0.01), SM d20: 1/22:2 lipid (β-coefficient 4.96; 95% CI 2.29 to 9.13; OR: 142.31; OR CI: 9.90 to 9.22^103; Wald's test: 2.93; p = 0.003); PG 20:3_22: 6 lipid (β-coefficient 2.52; 95% CI 1.13 to 4.49; OR: 12.37; OR CI: 3.10 to 89.27; Wald's test: 3.02; p = 0.002); PC O- 16: 1/16:0 lipid (β-coefficient 1.96; 95% CI -4.12 to 0.27; OR: 0.14; OR CI: 0.02 to 0.76; Wald's test: -2.05; p = 0.04). The prediction model characteristics of sperm maturation arrest, obtained during cross-validation in the positiveion mode composed: sensitivity 91%, specificity 85%; in negative-ion mode: sensitivity 75%; specificity 81%.Conclusions. Even though early stages of spermatogenesis are equally preserved in both fertile men and men with homogeneous sperm maturation arrest, the semen in the studied group of patients differed in its lipid profile. Patients with non-obstructive azoospermia, associated with meiosis arrest, may have unique lipidomic characteristics of seminal plasma, which in the future will make it possible to differentiate various variants of severe male infertility using non-invasive methods.

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CNV Hotspots in Testicular Seminoma Tissue and Seminal Plasma

Cancers ◽

10.3390/cancers14010189 ◽

2021 ◽

Vol 14 (1) ◽

pp. 189

Author(s):

Dora Raos ◽

Irena Abramović ◽

Miroslav Tomić ◽

Alen Vrtarić ◽

Tomislav Kuliš ◽

...

Keyword(s):

Seminal Plasma ◽

Life Quality ◽

Testicular Tissue ◽

Clinical Value ◽

Testicular Seminoma ◽

Aberrant Expression ◽

Before And After ◽

Number Variation ◽

Detection And Diagnosis ◽

Digital Polymerase Chain Reaction

Seminoma (SE) is the most frequent type of testicular tumour, affecting predominantly young men. Early detection and diagnosis of SE could significantly improve life quality and reproductive health after diagnosis and treatment. Copy number variation (CNV) has already been associated with various cancers as well as SE. In this study, we selected four genes (MAGEC2, NANOG, RASSF1A, and KITLG) for CNV analysis in genomic DNA (gDNA), which are located on chromosomes susceptible to gains, and whose aberrant expression was already detected in SE. Furthermore, CNV was analysed in cell-free DNA (cfDNA) from seminal plasma. Analysis was performed by droplet digital polymerase chain reaction (ddPCR) on gDNA from SE and nonmalignant testicular tissue. Seminal plasma cfDNA from SE patients before and after surgery was analysed, as well as from healthy volunteers. The CNV hotspot in gDNA from SE tissue was detected for the first time in all analysed genes, and for two genes, NANOG and KITLG it was reflected in cfDNA from seminal plasma. Although clinical value is yet to be determined, presented data emphasize a potential use of CNV as a potential SE biomarker from a liquid biopsy.

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