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

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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Cellular Therapy via Spermatogonial Stem Cells for Treating Impaired Spermatogenesis, Non-Obstructive Azoospermia

Cells ◽

10.3390/cells10071779 ◽

2021 ◽

Vol 10 (7) ◽

pp. 1779

Author(s):

Nesma E. Abdelaal ◽

Bereket Molla Tanga ◽

Mai Abdelgawad ◽

Sahar Allam ◽

Mostafa Fathi ◽

...

Keyword(s):

Stem Cells ◽

Male Infertility ◽

Cellular Therapy ◽

Antihypertensive Drugs ◽

Testicular Biopsy ◽

Testicular Tissue ◽

Spermatogonial Stem Cells ◽

Obstructive Azoospermia ◽

Promising Alternative ◽

Major Health Problem

Male infertility is a major health problem affecting about 8–12% of couples worldwide. Spermatogenesis starts in the early fetus and completes after puberty, passing through different stages. Male infertility can result from primary or congenital, acquired, or idiopathic causes. The absence of sperm in semen, or azoospermia, results from non-obstructive causes (pretesticular and testicular), and post-testicular obstructive causes. Several medications such as antihypertensive drugs, antidepressants, chemotherapy, and radiotherapy could lead to impaired spermatogenesis and lead to a non-obstructive azoospermia. Spermatogonial stem cells (SSCs) are the basis for spermatogenesis and fertility in men. SSCs are characterized by their capacity to maintain the self-renewal process and differentiation into spermatozoa throughout the male reproductive life and transmit genetic information to the next generation. SSCs originate from gonocytes in the postnatal testis, which originate from long-lived primordial germ cells during embryonic development. The treatment of infertility in males has a poor prognosis. However, SSCs are viewed as a promising alternative for the regeneration of the impaired or damaged spermatogenesis. SSC transplantation is a promising technique for male infertility treatment and restoration of spermatogenesis in the case of degenerative diseases such as cancer, radiotherapy, and chemotherapy. The process involves isolation of SSCs and cryopreservation from a testicular biopsy before starting cancer treatment, followed by intra-testicular stem cell transplantation. In general, treatment for male infertility, even with SSC transplantation, still has several obstacles. The efficiency of cryopreservation, exclusion of malignant cells contamination in cancer patients, and socio-cultural attitudes remain major challenges to the wider application of SSCs as alternatives. Furthermore, there are limitations in experience and knowledge regarding cryopreservation of SSCs. However, the level of infrastructure or availability of regulatory approval to process and preserve testicular tissue makes them tangible and accurate therapy options for male infertility caused by non-obstructive azoospermia, though in their infancy, at least to date.

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MicroRNA signature in spermatozoa and seminal plasma of proven fertile men and in testicular tissue of men with obstructive azoospermia

Andrologia ◽

10.1111/and.13503 ◽

2019 ◽

Vol 52 (2) ◽

Cited By ~ 1

Author(s):

Masood Abu‐Halima ◽

Valentina Galata ◽

Christina Backes ◽

Andreas Keller ◽

Mohamad Hammadeh ◽

...

Keyword(s):

Seminal Plasma ◽

Testicular Tissue ◽

Obstructive Azoospermia

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tRNA-Derived Fragments in Seminal Plasma Exosomes Are Non-invasive Biomarkers for Diagnosis of Non-obstructive Azoospermia With Spermatogenic Failure

10.21203/rs.3.rs-89955/v1 ◽

2020 ◽

Author(s):

Zuobin Zhu ◽

Xiaoxiao Han ◽

Ying Li ◽

Yao Zhu ◽

Zhenbei Li ◽

...

Keyword(s):

Seminal Plasma ◽

Predictive Accuracy ◽

Preoperative Evaluation ◽

Bioinformatic Analysis ◽

Testicular Tissue ◽

Obstructive Azoospermia ◽

Diagnostic Biomarker ◽

Testicular Sperm ◽

Non Invasive ◽

Qrt Pcr

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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Xenogeneic transplantation of human spermatogonia

Zygote ◽

10.1017/s0967199400000873 ◽

2000 ◽

Vol 8 (2) ◽

pp. 97-105 ◽

Cited By ~ 39

Author(s):

Marcos M. Reis ◽

Ming C. Tsai ◽

Peter N. Schlegel ◽

Miriam Feliciano ◽

Ricciarda Raffaelli ◽

...

Keyword(s):

Germ Cells ◽

Cell Types ◽

Testicular Biopsy ◽

Testicular Tissue ◽

Seminiferous Tubules ◽

Obstructive Azoospermia ◽

Spermatogenic Cells ◽

Immunodeficient Mice ◽

Xenogeneic Transplantation ◽

Immunological Rejection

In the last 3 years, several studies have shown that xenogeneic transplantation of rodent spermatogonia is feasible. The treatment of infertile patients with spermatogenic arrest using the injection of immature germ cells has yielded only poor results. We attempted to establish a complete spermatogenetic line in the testes of mutant aspermatogenic (W/Wv) and severe combined immunodeficient mice (SCID) transplanted with germ cells from azoospermic men. Spermatogenic cells were obtained from testicular biopsy specimens of men (average age of 34.3 ± 9 years) undergoing infertility treatment because of obstructive and non-obstructive azoospermia. Testicular tissue was digested with collagenase to promote separation of individual spermatogenic cells. The germ cells were injected into mouse testicular seminiferous tubules using a microneedle (40 μm inner diameter) on a 10 ml syringe. To assess the penetration of the cell suspension into the tubules, trypan blue was used as an indicator. Mice were maintained for 50 to 150 days to allow time for germ cell colonisation and development prior to them being killed. Testes were then fixed for histological examination and approximately 100 cross-sectioned tubules were examined for human spermatogenic cells. A total of 26 testicular cell samples, 16 frozen and 10 fresh, were obtained from 24 men. The origin of the azoospermia was obstructive (OA) in 16 patients and non-obstructive (NOA) in 8 patients. The concentration of spermatogenic cells in the OA group was 6.6 × 106 cells/ml, and 1.3 ? 106 cells/ml in the NOA group (p < 0.01). The different spermatogenic cell types were distributed equally in the OA samples, ranging from spermatogenia to fully developed spermatozoa, but in the NOA group the majority of cells were spermatogonia and spermatocytes. A total of 23 testes from 14 W/Wv mice and 24 testes from 12 SCID mice were injected successfully, as judged by the presence of spermatogenic cells in histological sections of testes removed immediately after the injection. However, sections from the remaining testes examined up to 150 days after injection showed tubules lined with Sertoli cells and xenogeneic germ cells were not found. The reason why the two strains of mouse used as recipients did not allow the implantation of human germ cells is probably due to interspecies specificity involving non-compatible cell adhesion molecules and/or immunological rejection.

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FSH levels and testicular volumes are associated with the severity of testicular histopathology in men with non-obstructive azoospermia

Journal of Assisted Reproduction and Genetics ◽

10.1007/s10815-021-02313-y ◽

2021 ◽

Author(s):

Parviz K. Kavoussi ◽

Kayla Hudson ◽

G. Luke Machen ◽

Maya Barsky ◽

Dan I. Lebovic ◽

...

Keyword(s):

Retrospective Chart Review ◽

Testicular Biopsy ◽

Testicular Volume ◽

Testicular Sperm Extraction ◽

Obstructive Azoospermia ◽

Sperm Retrieval ◽

Maturation Arrest ◽

Early Maturation ◽

Potential Association ◽

The Mean

Abstract Purpose The purpose of this study is to assess a potential association between FSH levels and testicular volumes with the severity of testicular histopathology on testicular biopsy in men with non-obstructive azoospermia (NOA) undergoing microdissection testicular sperm extraction (microTESE). Methods A retrospective chart review was performed from the electronic health records of men who underwent microTESE with NOA. Results Eighty-six men with NOA underwent microTESE with concomitant testicular biopsy for permanent section to assess the testicular cellular architecture. The histopathological patterns were categorized by severity indicating the odds of sperm retrieval into 2 categories. The unfavorable category included Sertoli cell only pattern and early maturation arrest (n = 50) and the favorable category included late maturation arrest and hypospermatogenesis patterns (n = 36). In the men with unfavorable histopathologic patterns, the mean FSH level was 22.9 ± 16.6 IU/L, and the mean testicular volume was 10.4 ± 6.0 cc. This was in comparison to men with favorable histopathologic patterns revealing a mean FSH level of FSH 13.3 ± 12.0 with a mean testicular volume of 13.3 ± 5.9 cc. There was a statistically significant higher FSH level in men with unfavorable histopathology than favorable (p = 0.004) as well as a significant smaller mean testicular volume in men with unfavorable histopathology (p = 0.029). Conclusions Higher serum FSH levels and smaller testicular volumes are associated with more severe testicular histopathological patterns in men with NOA.

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Confounding contaminants in mass spectrometric reaction monitoring

10.26434/chemrxiv.7488737.v1 ◽

2018 ◽

Author(s):

Gilian T. Thomas ◽

Landon MacGillivray ◽

Natalie L. Dean ◽

Rhonda L. Stoddard ◽

Lars Yunker ◽

...

Keyword(s):

Mass Spectrometry ◽

Mass Spectrometric ◽

Negative Ion ◽

Reaction Monitoring ◽

Schlenk Flask ◽

Dynamic Analyses ◽

Rubber Septa ◽

Negative Ion Mode ◽

Mode A

<p>Reactions carried out in the presence of rubber septa run the risk of additives being leached out by the solvent. Normally, such species are present at low enough levels that they do not interfere with the reaction significantly. However, when studying reactions using sensitive methods such as mass spectrometry, the appearance of even trace amounts of material can confuse dynamic analyses of reactions. A wide variety of additives are present in rubber along with the polymer: antioxidants, dyes, detergent, and vulcanization agents, and these are all especially problematic in negative ion mode. A redesigned Schlenk flask for pressurized sample infusion (PSI) is presented as a means of practically eliminating the presence of contaminants during reaction analyses.</p>

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Dopant-Enriched Nitrogen Gas for Enhanced Electrospray Ionization of Released Glycans in Negative Ion Mode

Analytical Chemistry ◽

10.1021/acs.analchem.1c00023 ◽

2021 ◽

Author(s):

Katarina Madunić ◽

Sander Wagt ◽

Tao Zhang ◽

Manfred Wuhrer ◽

Guinevere S.M. Lageveen-Kammeijer

Keyword(s):

Electrospray Ionization ◽

Negative Ion ◽

Nitrogen Gas ◽

Negative Ion Mode

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Decreased expression of DNA methyltransferases in the testes of patients with non-obstructive azoospermia leads to changes in global DNA methylation levels

Reproduction Fertility and Development ◽

10.1071/rd18246 ◽

2019 ◽

Vol 31 (8) ◽

pp. 1386 ◽

Cited By ~ 1

Author(s):

Fatma Uysal ◽

Gokhan Akkoyunlu ◽

Saffet Ozturk

Keyword(s):

Dna Methylation ◽

Male Infertility ◽

De Novo ◽

Biological Effects ◽

Testicular Biopsy ◽

Round Spermatid ◽

Dna Methyltransferases ◽

Global Dna Methylation ◽

Obstructive Azoospermia ◽

Spermatogenic Cells

DNA methylation plays key roles in epigenetic regulation during mammalian spermatogenesis. DNA methyltransferases (DNMTs) function in de novo and maintenance methylation processes by adding a methyl group to the fifth carbon atom of the cytosine residues within cytosine–phosphate–guanine (CpG) and non-CpG dinucleotide sites. Azoospermia is one of the main causes of male infertility, and is classified as obstructive (OA) or non-obstructive (NOA) azoospermia based on histopathological characteristics. The molecular background of NOA is still largely unknown. DNA methylation performed by DNMTs is implicated in the transcriptional regulation of spermatogenesis-related genes. The aim of the present study was to evaluate the cellular localisation and expression levels of the DNMT1, DNMT3A and DNMT3B proteins, as well as global DNA methylation profiles in testicular biopsy samples obtained from men with various types of NOA, including hypospermatogenesis (hyposperm), round spermatid (RS) arrest, spermatocyte (SC) arrest and Sertoli cell-only (SCO) syndrome. In the testicular biopsy samples, DNMT1 expression and global DNA methylation levels decreased gradually from the hyposperm to SCO groups (P<0.05). DNMT3A expression was significantly decreased in the RS arrest, SC arrest and SCO groups compared with the hyposperm group (P<0.05). DNMT3B expression was significantly lower in the RS arrest and SCO groups than in the hyposperm group (P<0.05). Although both DNMT1 and DNMT3A were localised in the cytoplasm and nucleus of the spermatogenic cells, staining for DNMT3B was more intensive in the nucleus of spermatogenic cells. In conclusion, the findings suggest that significant changes in DNMT expression and global DNA methylation levels in spermatogenic cells may contribute to development of male infertility in the NOA groups. Further studies are needed to determine the molecular biological effects of the altered DNMT expression and DNA methylation levels on development of male infertility.

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