Biomarkers Of Male Infertility Research Articles

Expression of exosomal microRNAs miR-34a and miR-210 in male infertility: relationship with morphokinetic parameters and sperm DNA fragmentation

Introduction. Infertility affects tens of millions of men and women across the globe. In approximately half of cases, male factors are the cause of infertility. In recent decades, there has been a significant decline in the quality of male ejaculate, which is characterized by reduced sperm concentration and motility. The insufficient diagnostic and prognostic value of routine semen analysis results highlights the challenge of developing effective diagnostic tools and searching for reliable biomarkers of male infertility. One of the most promising approaches may be assessing sperm microRNA expression.Objective. To study the role of sperm exosomal microRNAs miR-34a and miR-210 in the development of male infertility.Materials & methods. The retrospective study included 150 men aged 25 – 49 years; of these, 96 patients were diagnosed with idiopathic infertility. The comparison group consisted of 54 fertile men. To assess the structure and motility of sperm, the results of a standard ejaculate study (WHO, 2021) and computer data analysis using MMC Sperm (MMCSoft, St. Petersburg, Russia) software were used. The degree of DNA fragmentation was assessed using the TUNEL method. To analyze the expression of miR-34a and miR-210, quantitative real-time PCR was performed using the miRCURY LNA SYBR Green PCR Kit («Qiagen» GmbH, Hilden, Germany) and the Rotor-Gene Q PCR product detection system («Qiagen» GmbH, Hilden, Germany).Results. The study of ejaculate using the Computer Assisted Sperm Analysis System (CASA) method revealed a statistically significant relationship between the level of DNA fragmentation of sperm and indicators of the speed of their movement: rectilinear (VSL) (r = -0.522726; p < 0.01), curvilinear (VCL) (r = -0.499096; p < 0.01), along the middle path (VAP) (r = -0.429533; p < 0.01), as well as with the amplitude of lateral head displacement (ALH) (r = -0.294779, p < 0.01), the linearity of their curvilinear path (LIN) (r = -0.385796; p < 0.01), the degree of straight-line movements (STR) (r = -0.268248; p < 0.05) and their progressive mobility (r = -0.411547; p < 0.01). A study of the level of microRNA expression in sperm exosomes revealed a statistically significant decrease in its miRNA-34a pool (p = 0.0116). According to the Chaddock scale, the strength of the correlation between miR-210 expression and the effectiveness of ART programs was moderate (0.437993). The inverse relationship between miR-34a expression and IVF and ICSI results was weak (0.135314).Conclusion. The analysis of exosomal microRNA-34a and microRNA-210, which are involved in the regulation of spermatogenesis, reveals a direct correlation between their variations and changes in the kinetic and morphological parameters of gametes. It also indicates a relationship with the state of DNA fragmentation. These findings suggest varying levels of gene expression among infertile patients, men with proven fertility, and those undergoing assisted reproductive technology (ART) treatments, both with successful and repeated unsuccessful outcomes.

Sperm RNA quantity and PRM1, PRM2, and TH2B transcript levels reflect sperm characteristics and early embryonic development.

Spermatozoa have a highly complex RNA profile. Several of these transcripts are suggested as biomarkers for male infertility and contribute to early development. To analyze the differences between sperm RNA quantity and expression of protamine (PRM1 and PRM2) and testis-specific histone 2B (TH2B) genes, spermatozoa from 33 patients who enrolled in assisted reproduction treatment (ART) program were analyzed. Sperm RNA of teratozoospermic (T), oligoteratozoospermic (OT), and normozoospermic (N) samples was extracted, and the differences in transcript levels among the study groups were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). The correlations of total RNA per spermatozoon and the expression of the transcripts were evaluated in relation to sperm characteristics and preimplantation embryo development. The mean (±standard deviation) RNA amount per spermatozoon was 28.48 (±23.03) femtogram in the overall group and was significantly higher in the OT group than that in N and T groups. Total sperm RNA and gene expression of PRM1 and PRM2 genes were related to preimplantation embryo development and developmental arrest. Specific sperm characteristics were correlated with the expressions of PRM1, PRM2, or TH2B genes. We conclude that the sperm RNA amount and composition are important factors and might influence early embryonic development and also differ in different cases of male infertility.

Potential biomarker signatures in male infertility: integrative genomic analysis

BackgroundStudies have attributed 50% of infertility cases to male infertility, 15% of which is caused by idiopathic genetic factors. Currently, no specific biomarkers have been revealed for male infertility. Furthermore, research on genetic factors causing male infertility is still limited. As with other multifactorial genetic disorders, numerous risk loci for male infertility have been identified by genome-wide association studies (GWAS), although their clinical significance remains uncertain. Therefore, we utilized an integrative bioinformatics-based approach to identify biomarkers for male infertility. Bioinformatics analysis was performed using Open Targets Platform, DisGeNet, and GWAS Catalog. After that, the STRING database and the Cytoscape program were used to analyze protein–protein interaction. CytoHubba was used to determine the most significant gene candidates. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses were used to assess biological functions that correspond to the male infertility disease pathway.ResultsWe identified 305 genes associated with male infertility and highlighted 10 biological risk genes as potential biomarkers for male infertility such as TEX11, SPO11, SYCP3, HORMAD1, STAG3, MSH4, SYCP2, SYCE1, RAD21L1, and AMH. Of all the genes, we took the top three genes, namely, TEX11, SPO11, and SYCP3 as the genes that have the most potential as biomarkers.ConclusionsTEX11, SPO11, and SYCP3 are involved in meiosis and spermatogenesis. We propose that further research in regarding these genes in detecting male infertility.

Human circBOULE RNAs as potential biomarkers for sperm quality and male infertility.

Reliable molecular biomarkers to predict fertility remain scarce. The current study explored the potential of testis-specific circBOULE RNAs as biomarkers for male infertility and sperm quality. Using RT-PCR and RT-qPCR assays, we identified seven circular RNAs from the human BOULE gene in human sperm. We found that sperm circEx3-6 RNA exhibited a significantly decreased expression in asthenozoospermia while circEx2-6 and circEx2-7 expression decreased in teratozoospermia, compared with the controls. Furthermore, circEx2-6 expression exhibited a negative correlation with sperm DNA Fragmentation Index (DFI), and circEx2-7 levels were correlated with both fertilization and cleavage rates involving assisted reproductive technologies. Further functional analyses in a transgenic fly model lent support for the roles of circBOULE RNAs in sperm development and human fertility. Collectively, our findings support that sperm circBOULE RNAs may serve as diagnostic biomarkers for assessing sperm motility and DNA quality. Hence clinical application and significance of sperm circular RNAs in assisted reproductive technologies warrant further investigation.

T7 RNA polymerase-mediated rolling circle transcription and the CRISPR-Cas13a cascade reaction for sensitive and specific detection of piRNA.

The aberrant expression of piRNAs in germ cells is a potential cause of male infertility. Establishing diagnostic methods with highly specific biomarkers for male infertility is important for accurate diagnosis and treatment of male infertility. In this study, we proposed a novel method combining rolling circle transcription (RCT) and Cas13a techniques, which utilized the high amplification efficiency of RCT and the two different RNase activities possessed by Cas13a, establishing a highly sensitive and specific assay for male infertility-associated piRNA. First, a circular DNA template was synthesized by hybridizing linear ssDNA with the T7 promoter. The nick in the circular DNA was closed by T4 DNA ligase. In the presence of T7 RNA polymerase, the closed circular DNA produced tandemly repeated pre-crRNA. The RNase activity of Cas13a was used to process pre-crRNAs to form mature crRNA. Guided by crRNA, Cas13a specifically recognized piRNA and activated collateral activity. Activated Cas13a disaggregated thousands of fluorescent probes for each target RNA detected, resulting in powerful signal amplification. As a proof of concept, piR-hsa-14 was used as the validation target. The limit of detection was as low as 3.32 fM with a good linearity in the range of 100 fM to 50 pM. Recovery of piR-hsa-14 ranged from 91.33% to 112.63% in spiked recovery experiments using human serum samples. The results revealed that this method has the advantages of high sensitivity, sufficient accuracy and good reproducibility. We believe that this method could have a promising future as a potential tool for clinical diagnosis of male infertility.

Oxidative Stress, Telomere Length and Telomerase Activity in Spermatogenesis Disorders (Review of Scientific Activity).

The paper systematizes the available data on the study of oxidative stress, the relative length of telomeres, and telomerase activity in male infertility and disorders of spermatogenesis. The study of telomeres, the structures that protect chromosome ends and genome integrity, is of interest for researchers in various fields, from cell biology and epidemiology to ecology and evolutionary biology. The review includes our own data on the study of the relative length of telomeres, oxidative stress, and telomerase activity and reflects modern ideas about the importance of these structures both in the maintenance of genome stability during cell division and in gametogenesis and reproduction. Many studies indicate the role of oxidative stress in the pathogenesis of various diseases, including male infertility. In turn, studies of telomeres as a biomarker of male infertility are insufficient, and the results obtained are extremely controversial and require deeper knowledge about the mechanisms underlying the dynamics of telomere length.

Multi-omics study identifies that PICK1 deficiency causes male infertility by inhibiting vesicle trafficking in Sertoli cells

BackgroundInfertility affects approximately 10–15% of reproductive-age men worldwide, and genetic causes play a role in one-third of cases. As a Bin-Amphiphysin-Rvs (BAR) domain protein, protein interacting with C-kinase 1 (PICK1) deficiency could lead to impairment of acrosome maturation. However, its effects on auxiliary germ cells such as Sertoli cells are unknown.PurposeThe present work was aimed to use multi-omics analysis to research the effects of PICK1 deficiency on Sertoli cells and to identify effective biomarkers to distinguish fertile males from infertile males caused by PICK1 deficiency.MethodsWhole-exome sequencing (WES) was performed on 20 infertility patients with oligozoospermia to identify pathogenic PICK1 mutations. Multi-omics analysis of a PICK1 knockout (KO) mouse model was utilized to identify pathogenic mechanism. Animal and cell function experiments of Sertoli cell-specific PICK1 KO mouse were performed to verify the functional impairment of Sertoli cells.ResultsTwo loss-of-function deletion mutations c.358delA and c.364delA in PICK1 resulting in transcription loss of BAR functional domain were identified in infertility patients with a specific decrease in serum inhibin B, indicating functional impairment of Sertoli cells. Multi-omics analysis of PICK1 KO mouse illustrated that targeted genes of differentially expressed microRNAs and mRNAs are significantly enriched in the negative regulatory role in the vesicle trafficking pathway, while metabolomics analysis showed that the metabolism of amino acids, lipids, cofactors, vitamins, and endocrine factors changed. The phenotype of PICK1 KO mouse showed a reduction in testis volume, a decreased number of mature spermatozoa and impaired secretory function of Sertoli cells. In vitro experiments confirmed that the expression of growth factors secreted by Sertoli cells in PICK1 conditional KO mouse such as Bone morphogenetic protein 4 (BMP4) and Fibroblast growth factor 2 (FGF2) were decreased.ConclusionsOur study attributed male infertility caused by PICK1 deficiency to impaired vesicle-related secretory function of Sertoli cells and identified a variety of significant candidate biomarkers for male infertility induced by PICK1 deficiency.

Proteomic approach towards identification of seminal fluid biomarkers from individuals with severe oligozoospermia, cryptozoospermia and non-obstructive azoospermia: a pilot study.

Infertility becomes a global problem that affects to the same extent females and males. As reasons of male infertility can differ among individuals, the accurate diagnostics is essential for effective treatment. The most problematic both in diagnostics and in treatment are disturbances of spermatogenesis. Seminal fluid is rich in proteins that potentially can serve as markers for male infertility and among them, markers of spermatogenesis which are highly desired. To find biomarkers of spermatogenesis, we applied comparative proteomics using nano ultra performance liquid chromatography and tandem mass spectrometry (nanoUPLC-MS/MS) followed by single-sample Western blotting (WB) using seminal fluid samples from males with different types of infertility including non-obstructive azoospermia (NOA), cryptozoospermia (C) and severe oligozoospermia (SO). Then, the extensive survey on the identified proteins and their function in male reproductive system has been done. The proteomic approach has enabled to identified five seminal fluid proteins being potential markers of spermatogenesis disorders: ADGRG2, RAB3B, LTF, SLC2A3 and spermine synthase (SMS). Among them ADGRG2 seems to be strongly involved in male infertility. In addition, WB indicated that the distribution of LTF, SLC2A3 and SMS was not coherent among the individuals, especially in a group with NOA. Functional annotation analysis and search in proteomics databases revealed that vast majority of the proteins originated from extracellular environment. The presented data point out several proteins that potentially can become biomarkers of male infertility. The data suggest, however, different mechanisms behind the male infertility indicating that the etiology is more complex. We assume that recognition of these mechanisms may lead to the creation of specific protein panel helpful in the management of male infertility and therefore, further studies are required.

Seminal plasma untargeted metabolomic and lipidomic profiling for the identification of a novel panel of biomarkers and therapeutic targets related to male infertility.

Male infertility occurs approximately in about 50% of all infertility cases and represents a serious concern worldwide. Traditional semen analysis alone is insufficient to diagnose male infertility. Over the past two decades, advances in omics technologies have led to the widespread application of metabolomics profiling as a valuable diagnostic tool for various diseases and disorders. Seminal plasma represents a rich and easily accessible source of metabolites surrounding spermatozoa, a milieu that provides several indispensable nutrients to sustain sperm motility and fertilization. Changes of metabolic profiles in seminal plasma reflect male reproductive tract disorders. Here, we performed seminal plasma metabolomics and lipidomics profiling to identify a new pattern of biomarkers of male infertility. Seminal plasma samples from unfertile subjects (n = 31) and fertile controls (n = 19) were analyzed using an untargeted metabolomics/lipidomics integrated approach, based on Ultra-High-Pressure Liquid Chromatography-tandem Mass Spectrometry. Partial Least Squares-Discriminant Analysis showed a distinct separation between healthy fertile men and infertile subjects. Among the 15 selected candidate biomarkers based on Variable Importance in Projection scores, phosphatidylethanolamine (PE) (18:1; 18:1) resulted with the highest score. In total, 40 molecular species showed statistically significant variations between fertile and infertile men. Heat-map and volcano plot analysis indicated that acylcarnitines, phosphatidylserine (PS) (40:2) and lactate were decreased, while PE (18:1; 18:1), Phosphatidic acid (PA) (O-19:2; 18:1), Lysophosphatidylethanolamine (LPE) (O-16:1) and Phosphatidylcholine (PC) (O-16:2; 18:1)-CH3 were increased in the infertile group. The present study is the first one to analyze the metabolomics/lipidomics dysregulation in seminal plasma between fertile and infertile individuals regardless of sub-infertility condition. Association of several metabolites/lipids dysregulation with male infertility reinforced data of previous studies performed with different approaches. In particular, we confirmed significantly decreased levels of PS and carnitines in infertile patients as well as the positive correlation with sperm motility and morphology. If validated on a larger prospective cohort, the metabolite biomarkers of infertility in seminal plasma we identified in the present study might inform novel strategies for diagnosis and interventions to overcome male infertility.

The role of small non-coding RNAs (sncRNAs) in male infertility: A scoping review

Objective: To give a brief overview of the field of epigenetics and the potential predictive power that small non-coding RNA (sncRNA) may hold in relation to improving the treatment and diagnosis of male infertility. Methods: PRISMA-ScR was used as the scoping review guideline for this investigation. All article data here have been accessed from MEDLINE–PubMed, Science Direct, EBSCO, Scopus, Sage Journals, and Google Scholar. The terms "small non coding RNA, male, infertility, miRNA, sperm" were used in the search between 2015 and 2023. Results: The study comprised 35 publications in total. Several sncRNAs, miR-155, miR-16, miR-196, miR-525-3p, miR-891 were found to be effective in regulating the mechanism of spermatozoa processing in the infertility of men. sncRNA can be used as a biomarker of male infertility. Conclusions: sncRNAs can act as biomarkers for the diagnosis of reproductive diseases. Actually, by recognizing sncRNAs and their mechanisms, a new way to treat infertile men would be paved. The functional annotation of sncRNAs in spermatogenesis is still in its infancy but has enormous potential. This is despite the fact that many potential sncRNAs have been found to date with the use of cutting-edge technology and publicly accessible sncRNA annotation tools.

Seminal plasma testis expressed sequence (TEX)-101 as a biomarker for the qualitative assessment of male factor infertility: A case-control study

ObjectiveThe study was conducted to assess the role of seminal plasma Testis Expressed Sequence (TEX)-101 as a biomarker of male infertility. Study designThe study was conducted on 180 men (20–50 years) with 90 having abnormal semen reports as cases and 90 with normal reports as controls in a rural tertiary care center in Southern India over two years. After the enrolment of cases and control, semen samples were cryopreserved till the desired sample size was achieved and a biochemical test for TEX-101 was run using Human Testis-expressed Protein 101 ELISA Kit. The results of TEX-101 were compared between cases and controls and correlated with various semen parameters. Statistical analysis was done using SPSS software version 22.0, a p-value < 0.05 was considered statistically significant. ResultsMean ± SD age of all participants was 29.94 ± 4.91 years. Of 90 cases, 48.9% had asthenospermia, 24.4% oligoasthenospermia, 15.6% oligospermia, 11.1% azoospermia. A statistically significant difference was observed in mean values of seminal plasma TEX-101 between cases (1.45 ± 0.08 ng/mL) and controls (2.26 ± 0.18 ng/mL), p = 0.001. A significant correlation (p = 0.001) was found between seminal TEX-101, semen volume, sperm concentration, progressive motility, and morphology. The area under the Receiver Operating Characteristic curve of TEX-101 between cases and controls was 1.00 (p = 0.001), indicating TEX-101 as a potential biomarker for distinguishing men with abnormal semen parameters from those with normal semen parameters. At a cut-off value of 1.84 ng/mL, seminal plasma TEX-101 had a sensitivity, specificity, and negative and positive predictive values of 100% for male infertility prediction. ConclusionSeminal TEX-101 is a potential seminal biomarker and can be used in the qualitative assessment of male factor infertility.

Is telomere length a biomarker of sperm quality? A systematic review and meta-analysis of observational studies.

Telomeres are essential for the integrity of chromosome ends during cell division and their involvement in different processes linked to aging has been established. These chromosome components are involved in spermatogenesis and seem to play an important role in fertilization and embryo development. Telomere length is shortened with each cell division. Recently, short sperm telomere length has been proposed as a potential biomarker of male infertility. To conduct a systematic review and meta-analysis of studies exploring the association between spermatozoa and/or leukocyte telomere length with sperm quality parameters and different infertility conditions. A systematic review and meta-analysis was conducted with studies from Medline-PUBMED and Cochrane Library databases until May 2022. Eligible studies included cohort, cross-sectional and case-control studies, and telomere length in spermatozoa and/or leukocytes cells was defined as the exposure. Semen quality parameters or infertility conditions (e.g., oligozoospermia, asthenozoospermia, teratozoospermia, or other spermatogenic impairment combinations) were defined as the outcomes. Twenty-three observational studies were included. In the qualitative analysis, high heterogeneity was observed between studies regarding the associations between telomere length and semen parameters in different normozoospermic/fertile and oligozoospermic/infertile populations. In the meta-analysis, spermatozoa and leukocyte telomere length were shorter in infertile individuals than in fertile individuals (mean difference [95% confidence interval]: -1.43 [-1.66 to -1.21], p-value <0.001and -1.67 [-2.02 to -1.31], p-value <0.001, respectively). Moreover, in terms of sperm telomere length, these differences were also significant between individuals with a normal seminogram and individuals with a low quantity of spermatozoa in the ejaculate (-0.97 [-1.32, -0.61], p-value <0.001). The current systematic review and meta-analysis suggests the potential role of spermatozoa or leukocyte telomere length as a reliable biomarker of semen quality, which may help distinguish between infertility conditions beyond the routine semen analysis.

Targeted Amino Acids Profiling of Human Seminal Plasma from Teratozoospermia Patients Using LC-MS/MS.

Identifying the metabolome of human seminal plasma (HSP) is a new research area to screen putative biomarkers of infertility. This case-control study was performed on HSP specimens of 15 infertile patients with teratozoospermia (defined as normal sperm morphology < 4%) and 12 confirmed fertile normozoospermic men as the control group to investigate the seminal metabolic signature and whether there are differences in the metabolome between two groups. HSPs were subjected to LC-MS-MS analysis. MetaboAnalyst5.0 software was utilized for statistical analysis. Different univariate and multivariate analyses were used, including T-tests, fold change analysis, random forest (RF), and metabolite set enrichment analysis (MSEA). Teratozoospermic samples contained seventeen significantly different amino acids. Upregulated metabolites include glutamine, asparagine, and glycylproline, whereas downregulated metabolites include cysteine, γ-aminobutyric acid, histidine, hydroxylysine, hydroxyproline, glycine, proline, methionine, ornithine, tryptophan, aspartic acid, argininosuccinic acid, α-aminoadipic acid, and β-aminoisobutyric acid. RF algorithm defined a set of 15 metabolites that constitute the significant features of teratozoospermia. In particular, increased glutamine, asparagine, and decreased cysteine, tryptophan, glycine, and valine were strong predictors of teratozoospemia. The most affected metabolic pathways in teratozoospermic men are the aminoacyl-tRNA, arginine, valine-leucine, and isoleucine biosynthesis. Altered metabolites detected in teratozoospermia were responsible for various roles in sperm functions that classified into four subgroups as follows: related metabolites to antioxidant function, energy production, sperm function, and spermatogenesis. The altered amino acid metabolome identified in this study may be related to the etiology of teratozoospermia, and may provide novel insight into potential biomarkers of male infertility for therapeutic targets.

PUFAs and Their Derivatives as Emerging Players in Diagnostics and Treatment of Male Fertility Disorders.

About 15% of couples worldwide are affected by infertility, with the male factor responsible for approximately 50% of reproductive failures. Male fertility can be influenced by various factors, including an unhealthy lifestyle and diet, often associated with oxidative stress. These changes are frequently the reason for spermatozoan dysfunction, malformations, and lowered count. However, sometimes even with proper semen parameters, fertilization does not occur, and this is referred to as idiopathic infertility. Of particular importance may be molecules contained in the spermatozoan membrane or seminal plasma, such as polyunsaturated fatty acids, including omega-3 (docosahexaenoic and eicosapentaenoic acids) and omega-6 (arachidonic acid) fatty acids and their derivatives (prostaglandins, leukotrienes, thromboxanes, endocannabinoids, isoprostanes), which are vulnerable to the effects of oxidative stress. In the present review, we discuss the influence of these molecules on human male reproductive health and its possible causes, including disrupted oxidative-antioxidative balance. The review also discusses the potential use of these molecules in the diagnostics and treatment of male infertility, with a particular focus on the innovative approach to isoprostanes as biomarkers for male infertility. Given the high occurrence of idiopathic male infertility, there is a need to explore new solutions for the diagnosis and treatment of this condition.

Genetic alterations in azoospermia patients may reveal potential biomarkers for male infertility: A bioinformatic study

Background/Aim: Azoospermia is defined as the absence of sperm in semen and is one of the most common causes of male infertility, with a prevalence of 10-15% in infertile men. Conventional methods for semen analysis do not provide a clear understanding of the etiology of azoospermia. Although testicular biopsy may exclude obstructive cases, non-obstructive azoospermia (NOA) treatment is limited due to a limited understanding of the underlying molecular mechanisms. Analysis of genetic alterations in azoospermia patients compared to the fertile population may be a valuable tool for determining diagnostic biomarkers for male infertility. This study aims to use bioinformatic tools to determine the top candidates in certain pathways altered in azoospermia. Methods: Expression data (GSE108886) of the differential testicular transcriptome in patients with NOA was selected from the Gene Expression Omnibus (GEO) database. Testicular RNA was harvested from azoospermia patients (n=11) and healthy controls (n=1, pooled sample). The differentially expressed genes (DEGs) were examined using GEO2R software. Biological pathways were identified through the Kyoto Encyclopedia of Genes and Genomes (KEGG). Construction of the protein network and detection of hub genes were conducted in the STRING database. Data validation was performed via ELISA assay for the FOXO3 gene in obstructive and NOA patients. Significance was set at P-value &lt;0.05. Results: In NOA patients, 2115 genes were upregulated, and 1753 genes were downregulated compared to the control group. Ninety-one genes involved in spermatogenesis were downregulated. KEGG analysis revealed that the glucagon signaling, AMPK signaling, insulin and estrogen signaling, and oocyte meiosis pathways were upregulated, while the regulation of actin cytoskeleton, MAPK signaling pathway, focal adhesion, and chemical carcinogenesis – reactive oxygen species pathways were downregulated. Downstream genes with the highest score were PSMA4, PSMA6, PSMC1, PSME4, and UBA52, which are responsible for the ubiquitin-dependent protein degradation. The top hub genes with increasing expression were RPS18, RPS2, and RPS4X Conclusion: Although hub genes selected within the altering pathways may serve as a diagnostic tool for NOA, further validation of the presented data is necessary, as protein-protein interactions may not reflect alterations in gene expression in vivo.

Commentary: Cancer-testis antigen lactate dehydrogenase C4 as a novel biomarker of male infertility and cancer.

GENERAL COMMENTARY article Front. Oncol., 06 February 2023Sec. Genitourinary Oncology Volume 13 - 2023 | https://doi.org/10.3389/fonc.2023.1115620

Sperm telomere length as a novel biomarker of male infertility and embryonic development: A systematic review and meta-analysis.

Telomeres have an essential role in maintaining the integrity and stability of the human chromosomal genome and preserving essential DNA biological functions. Several articles have been published on the association of STL with male semen parameters and clinical pregnancy. The results, however, are either inconclusive or inconsistent. Therefore, this meta-analysis aimed to systematically assess the accuracy and clinical value of sperm telomere length (STL) as a new marker for diagnosing male infertility and predicting the quality of embryonic development. We performed a comprehensive systematic search for relevant publications in PubMed, the Cochrane Library, Web of Science, Embase, Scopus, and Ovid, from database build to August 2022. All experimental studies exploring the association of STL with male semen quality, male infertility, or embryonic development were included. Overall, Twelve prospective observational cohort studies (1700 patients) were eligible for inclusion in the meta-analysis. The meta-analysis showed a positive linear correlation between STL and semen parameters. The optimal cut-off value for STL diagnosing male infertility was 1.0, with a sensitivity and specificity of 80%. Regarding STL and embryonic development, the clinical pregnancy rate was associated with longer STL, and there was no significant difference between the two groups regarding fertilization rate. Our study showed that STL has good diagnostic and predictive value for male fertility and clinical pregnancy and could be used as a new biomarker for diagnosing male infertility and predicting embryonic development. https://www.crd.york.ac.uk/PROSPERO/, identifier CRD42022303333.

Cancer-testis antigen lactate dehydrogenase C4 as a novel biomarker of male infertility and cancer.

A unique lactate dehydrogenase (LDH) isoenzyme designated as lactate dehydrogenase C4 (LDH-C4) is found in mammalian mature testis and spermatozoa. Thus far, LDH-C4 has been well studied with regard to its gene and amino acid sequences, structure, biological properties, and peptide synthesis. Accumulating evidence has shown that LDH-C4 is closely related to spermatic energy metabolism and plays a critical role in sperm motility, capacitation, and fertilization. Defects in the catalytic activity of LDH-C4 are key to pathophysiological abnormalities underlying infertility. LDH-C4 was originally thought to be present only in mature testis and spermatozoa; however, recent studies have implicated LDH-C4 as a cancer-testis antigen (CTA), owing to its aberrant transcription in a broad spectrum of human neoplasms. This review highlights the recent findings on LDH-C4 with particular emphasis on its role in male infertility and tumors.

PMON255 Inflammation and Oxidative Stress in Seminal Plasma: Role of Free Light Chains of Immunoglobulins and Antioxidants as Biomarkers in Male Infertility

Abstract Oxidative and inflammatory damage are underlying mechanisms in various conditions related to male infertility, including varicocele. Free light chains of immunoglobulins (FLCs) are considered markers of low-grade inflammation in various conditions. Nowadays it is known that they may play enzymatic and proteolytic activities; they may activate complement cascade and mast cells, with crucial role in the development of contact sensibility; they may also inhibit the autonomous signaling ability of the B-cell receptor. Their detection in biological fluids may be useful as biomarker of inflammation or altered immune response. Coenzyme Q10 (CoQ10), a lipidic antioxidant and component of the mitochondrial respiratory chain, is involved in spermatozoa energy metabolism and motility. Therefore, we aimed to evaluate FLCs seminal levels in patients with varicocele in comparison to control subjects and to correlate them with CoQ10 and Total Antioxidant Capacity (TAC) in human semen. A total of 65 patients were enrolled, aged 18-50 years. Semen analysis was performed; patients were divided into three groups: controls (CTRL), 12 normozoospermic patients aged 34 (33-41) years; varicocele (VAR), 29 patients aged 33 (26-37) years; idiopathic (IDIO), 24 oligo-, astheno- and oligoasthenozoospermic patients aged 37 (33.5-40.5) years. FLCs were assayed by turbidimetric method; CoQ10 by HPLC; TAC by spectrophotometric method. Λ FLCs showed a trend towards higher levels in VAR vs CTRL and IDIO. VAR showed a trend towards lower κ FLCs levels vs the other two groups. Comparing κ/λ ratio VAR showed significantly lower levels vs CTRL and IDIO (Mean±SEM 3.18±0.75; 8.28±1.71; 8.90±2.94, respectively; p&amp;lt;0.05). CoQ10 seminal levels showed higher levels in VAR and IDIO compared to CTRL (57.7±4.13; 65.50±6.95 and 38.30±3.35 mcg/ml, respectively; p&amp;lt;0.05) No different TAC values were found between groups. Our data confirm lower levels of κ/λ ratio in VAR as previously reported and suggest a possible application as clinical biomarkers for male infertility. Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

Qualitative and quantitative assessment of sperm miRNAs identifies hsa-miR-9-3p, hsa-miR-30b-5p and hsa-miR-122-5p as potential biomarkers of male infertility and sperm quality

BackgroundIn contrast with the preceding stages of the germ cells, spermatozoa are unusually rich in small non-coding RNAs in comparison to the coding RNAs. These small RNAs may have had an essential role in the process of spermatogenesis or may have critical roles in the post-fertilization development. Sporadic efforts have identified a few differentially expressed miRNAs in infertile individuals, which do not replicate in other studies.MethodsIn order to identify miRNAs signatures of infertility or poor sperm quality, we compared miRNA differential expression data across nine datasets, followed by their analysis by real-time PCR in a case–control study. This was followed by the validation of potential biomarkers in yet another set of cases and controls. For this, total RNA was isolated from 161 sperm samples. miRNA expression levels in infertile cases and fertile controls were measured using TaqMan real-time PCR. Meta-analyses of two miRNAs (hsa-miR-9-3p and hsa-miR-122-5p) were performed using Comprehensive Meta‐Analysis Software (version 2). All statistical analyses were performed with the help of GraphPad Prism Software (version 8).ResultsLiterature search identified seven miRNAs (hsa-let-7a-5p, hsa-miR-9-3p, hsa-miR-22-5p, has-miR-30b-5p, hsa-miR-103-3p, hsa-miR-122-5p and hsa-miR-335-5p) showing consistent dysregulation in infertility across a minimum of four studies. In the discovery phase, six miRNAs showed strong association with infertility with four (hsa-miR-9-3p, hsa-miR-30b-5p, hsa-miR-103-3p and hsa-miR-122-5p) showing consistent differential regulation across all sub-groups. Receiver operating characteristic (ROC) curve analysis showed that the area under curve of > 0.75 was achieved by three (hsa-mir-9-3p, hsa-miR-30b-5p and hsa-miR-122-5p) miRNAs. In the validation phase, these three miRNAs showed consistent association with infertility (hsa-mir-9-3p, hsa-miR-30b-5p, and hsa-miR-122-5p). Meta-analysis on hsa-miR-122-5p showed its significant quantitative association with infertility [Hedge’s g = -2.428, p = 0.001 (Random effects)].ConclusionsThree miRNAs (hsa-miR-9-3p, hsa-miR-30b-5p and hsa-miR-122-5p) have strong linkage with infertility and a high potential as sperm quality biomarkers.