Subgroups Of Infertile Men Research Articles

Denosumab stimulates spermatogenesis in infertile men with preserved Sertoli cell capacity

Sperm production depends on proper Sertoli-germ cell interaction, and we hypothesized that receptor activator of nuclear factor κB ligand (RANKL) activity in Sertoli cells may influence spermatogenesis. Treatment with the RANKL inhibitor denosumab, normally used to treat osteoporosis, increased testicular weight, inhibin B, and germ cell proliferation in exvivo testis cultures and invivo in a humanized RANKL mouse. The effect on germ cell proliferation was positively associated with baseline serum concentrations of anti-müllerian hormone (AMH). In accordance, denosumab increased germ cell proliferation in exvivo human testis cultures with low/moderate but not severe impairment of Sertoli cell function. In a placebo-controlled randomized clinical trial, denosumab had no effect on semen quality but increased sperm concentration in a subgroup of infertile men with serum AMH ≥38 pmol/L at baseline. In conclusion, high serum AMH may increase the probability of a beneficial response to denosumab treatment in infertile men, thus suggesting a possible venue for precision medicine in male infertility.

Seminal Histone Deacetylase, Fructose and Serum Reproductive Hormones as Diagnostic Marker in Sub-Groups of Infertile Males

Accurate diagnosis of the cause of infertility assists in the choice of treatment modalities and amelioration of the associated psychosocial problems. The research was carried out using 75 infertile males and 75 males with proven fertility as controls. The anthropometrics (weight, height) were measured and body mass index (BMI) computed. Venous blood was collected from each participant, allowed to clot, and centrifuged to obtain the serum which was analysed for testosterone, follicle stimulating hormone (FSH) and luteinizing hormone (LH) concentrations. Semen was collected by masturbation and analysed for sperm quality, seminal fructose concentration, and histone deacetylase (HDAC) activities. A non-significant difference (P>0.05) in weight, height, and BMI; a significant increase (P<0.05) in ejaculated volume, serum LH, FSH, seminal fructose concentrations and HDAC activities; and a decrease in sperm count, sperm motility, serum testosterone concentration was observed among subgroups of infertile men. A direct and significant correlation exists between seminal HDAC activities and fructose concentration. Also, an inverse non-significant correlation exists between HDAC activities and spermatozoa motility. Base on the result obtained from this study, it can be concluded that measuring seminal fructose and HDAC activities in addition to routine biochemical and biophysical parameters will assist in diagnostic work up in subgroups of male infertility.

P-102 Inhibition of RANKL increases the number of motile sperm in a subgroup of infertile men

Abstract Study question Does inhibition of RANKL increase semen quality in infertile men? Summary answer Overall, inhibition of RANKL (Denosumab) did not affect semen quality. However, in a subgroup of men with high serum AMH sperm production was significantly increased. What is known already Currently, there is no treatment available for male infertility, but studies in animal and preclinical human data have suggested a role for RANKL in male reproduction. Study design, size, duration A single-center, double-blinded, randomized clinical trial in 100 infertile men randomly assigned (1:1) to s.c. injection with 60 mg/mL Denosumab or s.c. saline injection 1 mL (0.9%) (placebo). The primary endpoint was changes in semen production, though the study was designed as an explorative study, with the aim of investigating the profile of which infertile men who would have a beneficial treatment outcome of Denosumab. Participants/materials, setting, methods All participants were infertile men referred to our andrological clinic due to impaired semen quality. Blood and semen samples were collected at baseline and after one and two cycles of spermatogenesis on day 80 and day 160, respectively. Main results and the role of chance There were no differences in semen quality or reproductive hormones at day 80 or day 160 between men treated with Denosumab or placebo. However, men with a serum AMH ≥ 40 pmol/L had a significant increase in total motile sperm (143% (SD 116%); p = 0.011), progressive motile sperm (133% (SD 100%); p = 0.039) and a borderline increase in sperm concentration (64% (SD 95%); p = 0.056) at day 80 compared to baseline. Further stratifying men according to both serum AMH (≥ 40 pmol/L) and serum RANKL (highest 50%), showed a significantly higher increase in sperm concentration (107% (SD 111%) vs. -8% (SD 68%); p = 0.009), total motile sperm (256% (SD 270%) vs. 31% (SD 125%); p = 0.039) and progressive motile sperm (191% (SD 194%) vs. 18% (SD 134%) p = 0.013) at day 80 in men treated with Denosumab compared to placebo treated men. Furthermore, serum FSH (-28% (SD 33%) vs. 0%, (SD 22%); p = 0.041) and serum LH (-29% (SD 28%) vs. 20% (SD 51%), p = 0.048) were significantly lower and testosterone/LH ratio (50% (SD 51%) vs. -7% (SD 43%); p = 0.024) significantly higher at day 80 in men treated with Denosumab compared to placebo treatment. Limitations, reasons for caution This is a report from a randomized clinical trial, with an explorative design. Randomized clinical trials in a predefined group of infertile men with high serum AMH, are needed to explore if inhibition of RANKL improves semen quality and pregnancy rate in infertile couples with male factor infertility. Wider implications of the findings The increase in sperm production after Denosumab treatment in infertile men with preserved Sertoli cell function needs to be validated, in a trial selecting men based on serum AMH. Ultimately, Denosumab may be a potential breakthrough in treating male infertility at least in some infertile men with high serum AMH. Trial registration number NCT03030196

Inhibition of RANKL in the testis increased the number of motile sperm in a sub-group of infertile men with preserved Sertoli cell function

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Testicular microdissection following failed sperm aspiration: a single-center experience.

Azoospermia is a cause of infertility in a subgroup of infertile men. Sperm retrieval techniques including testicular sperm aspiration (TESA) and microscopic testicular sperm extraction (mTESE) are widely used. In this study, we have reviewed our findings regarding mTESE performed following a negative TESA outcome. This is a retrospective chart review study that included 41 infertile patients who underwent mTESE after a negative TESA outcome. Charts were reviewed for demographic data, type of infertility, and type of azoospermia. Hormone level analysis was done for follicle-stimulating hormone, luteinizing hormone, and testosterone. Testicular volume was estimated by ultrasound. The study included 41 patients who underwent mTESE following a negative TESA outcome. Most patients had primary infertility (n = 32; 78%). Of the 41 patients, four had a previous history of either TESE or orchidopexy, and two had a history of varicocelectomy before the recent percutaneous TESA procedure. There was no significant association between sperm retrieval and the different surgical procedures that had been performed. Of the 41 patients, 27 had positive sperm retrieval by mTESE with a success rate of 65.9%. The positive sperm retrieval rate of mTESE performed following a negative TESA outcome was reasonable (65.9%). No significant correlations were identified with all variables studied.

Epigenetic landscape of testis specific histone H2B variant and its influence on sperm function

BackgroundBiological relevance of the major testis specific histone H2B variant (TH2B) in sperm is not fully understood. Studies in TH2A/TH2B double knockout male mice indicate its role in chromatin compaction and male fertility. Additionally, the presence of TH2B and TH2A reportedly generates more dynamic nucleosomes, leading to an open chromatin structure characteristic of transcriptionally active genome. Given that mature human sperm are transcriptionally and translationally inactive, the presence of TH2B in mature sperm is intriguing. To address its role in sperm, we investigated the genome-wide localization of TH2B in sperm of fertile men.ResultsWe have identified the genomic loci associated with TH2B in fertile human sperm by ChIP-seq analysis. Bioinformatic analysis revealed ~ 5% sperm genome and 5527 genes to be associated with TH2B. Out of these 105 (1.9%) and 144 (2.6%) genes showed direct involvement in sperm function and early embryogenesis, respectively. Chromosome wide analysis for TH2B distribution indicated its least distribution on X and Y chromosomes and varied distribution on autosomes. TH2B showed relatively higher percentage of gene association on chromosome 4, 18, 3 and 2. TH2B enrichment was more in promoter and gene body region. Gene Ontology (GO) analysis revealed signal transduction and associated kinase activity as the most enriched biological and molecular function, respectively. We also observed the enrichment of TH2B at developmentally important loci, such as HOXA and HOXD and on genes required for normal sperm function, few of which were validated by ChIP-qPCR. The relative expression of these genes was altered in particular subgroup of infertile men showing abnormal chromatin packaging. Chromatin compaction positively correlated with sperm- motility, concentration, viability and with transcript levels of PRKAG2 and CATSPER B.ConclusionChIP-seq analysis of TH2B revealed a putative role of TH2B in sperm function and embryo development. Altered expression of TH2B associated genes in infertile individuals with sperm chromatin compaction defects indicates involvement of TH2B in transcriptional regulation of these genes in post meiotic male germ cells. This altered transcriptome may be a consequence or cause of abnormal nuclear remodeling during spermiogenesis.

To study of apoptotic gene expression through hypothalamus-pituitary-gonadal axis in

To investigate the apoptotic gene expression of Bcl-2, Cytochrome C, Caspase and procaspase with male hormone profile in infertile subjects for their relationship to sperm quality and cell death parameters. We undertook gene expression on a total of 400 individuals, including 100 fertile donors as controls and three subgroups of infertile men, normozoospermic (idiopathic unexplained; n=100), oligozoospermic (n=100) and asthenozoospermic (n=100). These participants were selected from Departments of Urology, King George’s Medical University, Lucknow, India. We used ELISA, quantitative real time PCR (qPCR) with lightCycler Fast Start DNA PLUS SybrGreen kit for IL-6 and TNF alpha, Bcl-2, Cytochrome C, Caspase and procaspase mRNA and their relation to male fertility. We found decreased sperm motion kinetics and altered male hormone profile was associated with decreased Bcl-2 and procaspase expression and increased of cytochrome c expression was significantly increased in the oligozoospermic and asthenozoospermic infertile subjects compared to healthy fertile subjects. Male hormones, Bcl-2, Cytochrome C and Caspase gene expression were altered in men with impaired fertility possibly via their associations with sperm count, motility and morphology.

Cytokines in the blood and semen of infertile patients.

Cytokines have been important mediators of the immunity and can be involved in numerous processes in the male genital tract including acting as immunomodulatory elements within the male gonad.The aims of this study were: 1) to detect pro- and anti-inflammatory cytokine levels in the control group and subgroups of infertile men; and 2) to set up the practical recommendations concerning determination of cytokine levels for the male infertility diagnosis.Observations were performed in a group of 82 men: healthy controls (n = 27) and infertile patients (n = 55). The male infertility group was further subdivided into patients with: varicocele (n = 22), idiopathic infertility (n = 13) and partners of couples with recurrent spontaneous abortion (RSA; n = 20). Semen analysis was determined following WHO criteria. The cytokine interleukin 1β (IL-1β), IL-6, IL-10, IL-18; tumor necrosis factor α (TNF-α), interferon g (IFN-g) and transforming growth factor β1 (TGF-β1) contents in serum and seminal plasma were determined by quantitative ELISA.An interesting marker of male infertility appears to be TGF-β1 (blood) significantly elevated in idiopathically infertile males and in the RSA group. Besides elevated TGF-β1 in a group of idiopathic infertility significantly elevated IL-10, IL-18, IFN-g (blood) and statistically decreased IL-1β while increased IFN-g were revealed in seminal plasma compared to healthy controls.We may postulate novel cytokine micropatterns for patients with different background of infertility. Therefore, circulating cytokines: IL-1β, IL-10, IL-18, TGF-β1, IFN-g and IL-1β, IFN-g and TGF-β1 in seminal plasma should be extended in evaluation of specific types of male infertility.

Testicular biopsy: clinical practice and interpretation

Testicular biopsy was considered the cornerstone of male infertility diagnosis for many years in men with unexplained infertility and azoospermia. Recent guidelines for male infertility have limited the indications for a diagnostic testicular biopsy to the confirmation of obstructive azoospermia in men with normal size testes and normal reproductive hormones. Nowadays, testicular biopsies are mainly performed for sperm harvesting in men with non-obstructive azoospermia, to be used for intracytoplasmic sperm injection. Testicular biopsy is also performed in men with risk factors for testicular malignancy. In a subgroup of infertile men, there is an increased risk for carcinoma in situ of the testis, especially in men with a history of cryptorchidism and testicular malignancy and in men with testicular atrophy. Ultrasonographic abnormalities, such as testicular microlithiasis, inhomogeneous parenchyma and lesions of the testes, further increase the risk of carcinoma in situ (CIS) in these men. For an accurate histological classification, proper tissue handling, fixation, preparation of the specimen and evaluation are needed. A standardized approach to testicular biopsy is recommended. In addition, approaches to the detection of CIS of the testis testicular immunohistochemistry are mandatory. In this mini-review, we describe the current indications for testicular biopsies in the diagnosis and management of male infertility.

The prevalence of chromosomal abnormalities in subgroups of infertile men

The prevalence of chromosomal abnormalities is assumed to be higher in infertile men and inversely correlated with sperm concentration. Although guidelines advise karyotyping infertile men, karyotyping is costly, therefore it would be of benefit to identify men with the highest risk of chromosomal abnormalities, possibly by using parameters other than sperm concentration. The aim of this study was to evaluate several clinical parameters in azoospermic and non-azoospermic men, in order to assess the prevalence of chromosomal abnormalities in different subgroups of infertile men. In a retrospective cohort of 1223 azoospermic men and men eligible for ICSI treatment, we studied sperm parameters, hormone levels and medical history for an association with chromosomal abnormalities. The prevalence of chromosomal abnormalities in the cohort was 3.1%. No association was found between chromosomal abnormalities and sperm volume, concentration, progressive motility or total motile sperm count. Azoospermia was significantly associated with the presence of a chromosomal abnormality [15.2%, odds ratio (OR) 7.70, P < 0.001]. High gonadotrophin levels were also associated with an increased prevalence of chromosomal abnormalities (OR 2.96, P = 0.013). Azoospermic men with a positive andrologic history had a lower prevalence of chromosomal abnormalities than azoospermic men with an uneventful history (OR 0.28, P = 0.047). In non-azoospermic men, we found that none of the studied variables were associated with the prevalence of chromosomal abnormalities. We show that the highest prevalence of chromosomal abnormalities is found in hypergonadotrophic azoospermic men with an uneventful andrologic history.

Profiling spermatogenic failure in adult testes bearing Sox9-deficient Sertoli cells identifies genes involved in feminization, inflammation and stress

BackgroundSox9 (Sry box containing gene 9) is a DNA-binding transcription factor involved in chondrocyte development and sex determination. The protein's absence in testicular Sertoli nurse cells has been shown to disrupt testicular function in adults but little is known at the genome-wide level about molecular events concomitant with testicular break-down.MethodsTo determine the genome-wide effect on mRNA concentrations triggered by the absence of Sox9 in Sertoli cells we analysed adult testicular tissue from wild-type versus mutant mice with high-density oligonucleotide microarrays and integrated the output of this experiment with regulatory motif predictions and protein-protein network data.ResultsWe report the genome-wide mRNA signature of adult testes lacking Sox9 in Sertoli cells before and after the onset of late spermatogenic failure as compared to fertile controls. The GeneChip data integrated with evolutionarily conserved Sox9 DNA binding motifs and regulatory network data identified genes involved in feminization, stress response and inflammation.ConclusionsOur results extend previous observations that genes required for female gonadogenesis are up-regulated in the absence of Sox9 in fetal Sertoli cells to the adult stage. Importantly, we identify gene networks involved in immunological processes and stress response which is reminiscent of a phenomenon occurring in a sub-group of infertile men. This suggests mice lacking Sox9 in their Sertoli cells to be a potentially useful model for adult human testicular failure.

Inhibin and steroid response to testicular stimulation with pure FSH (Metrodin) in infertile men with unilateral cryptorchidism.

Static measurements of immunoreactive inhibin have proven of little relevance in the diagnosis of testicular disorders. Dynamic evaluation of the inhibin secretory reserve might detect a specific Sertoli cell defect in a subgroup of infertile men. We compared the response of inhibin and steroids to an intravenous injection of pure FSH (Metrodin, Serono, 300 IU) in 13 infertile men with unilateral cryptorchidism to that in eight normal fertile men. Blood was aspirated before, 24, 48, and 72 h after the FSH injection. Two subgroups of patients with unilateral cryptorchidism were detected: those who responded by secreting inhibin in a pattern similar to normal men (seven patients), and those who responded poorly or not at all (six patients). The presumed cause of this difference is a defect of Sertoli cell reserve function due to a combination of insults to the testes, and not to cryptorchidism itself. The difference in response to FSH cannot be predicted from semen analysis nor from static hormone measurements. Overall, inhibin levels correlated significantly with the serum concentrations of FSH (r = -0.36, P < 0.05), testosterone (r = 0.37, P < 0.05), and 17-hydroxyprogesterone (r = 0.66, P < 0.001). It is concluded that, in infertile men with unilateral cryptorchidism, Stimulation of Sertoli cells by FSH can identify a subgroup of patients with Sertoli cell malfunction involving inhibin synthesis.

The present and future state of hormonal treatment for male infertility.

Although male factors contribute to over half of all cases of infertility, most infertile men are described as 'idiopathic oligo/asthenozoospermic' rather than diagnosed precisely; hence, specific medical treatment is not possible. One uncommon but treatable cause of male infertility is gonadotrophin deficiency in which gonadotrophin replacement therapy is highly effective at inducing spermatogenesis and fertility. Hormonal therapy is a logical approach for empirical drug therapy given the fundamental role of hormonal regulation in spermatogenesis. However, treatment with GnRH analogues, gonadotrophins, androgens, anti-estrogens, aromatase inhibitors, growth hormone- and prolactin-suppressing drugs is ineffective in unselected infertile men. Prolonged high-dose glucocorticoid therapy for sperm autoimmunity may improve pregnancy rates modestly, but the risks are generally unacceptable compared with IVF or ICSI. For these reasons, modern reproductive technologies, notably ICSI/IVF, have become the de-facto standard empirical treatment of male infertility, despite involving significant though infrequent risks to the fetus and mother. There remains a potential for hormonal methods to improve sperm quality or ultrastructure in subgroups of infertile men more responsive to hormonal manipulation or using novel protein or gene-targeted therapies or biochemical approaches based on post-hormonal receptor mechanisms that stimulate spermatogenesis. How such novel hormonal methods will develop in conjunction with improved ICSI/IVF or cloning technologies, and the potential role of adjunctive hormonal therapy remains to be clarified.

Generation of reactive oxygen species in subgroups of infertile men

The capacity to generate reactive oxygen species (ROS), both basally and after stimulation with the calcium ionophore A23187, was examined in the motile fraction of sperm isolated after swim-up from the semen of 10 naturally fertile men and three groups of infertile patients. The latter included: (1) men with a non-bacterial inflammation of the genital tract (n = 10); (2) men unable to impregnate their partners during an intra-uterine insemination programme (IUI) (n = 8) and their matched controls (n = 6); and (3) men with hypogonadotrophic hypogonadism (HH) who remained infertile after induction of spermatogenesis with gonadotrophin or gonadotrophin-releasing hormone therapy (n = 3) and their matched controls (n = 3). The levels of ROS production were elevated in the sperm of some infertile men with inflammation of the genital tract compared to those found in 10 naturally fertile men. In addition, sperm from those patients who remained infertile after an IUI programme produced higher amounts of ROS compared to their control group who became fertile. Similarly, the production of ROS by sperm from three patients with HH who remained infertile was significantly higher than those of the three men who became fertile. These data suggest that an excessive production of ROS by sperm may explain some cases of idiopathic male infertility.

LDH Isoenzymes in Semen of Infertile Men

LDH isoenzymes (LDH1, LDH2, LDH3, LDH4, LDH5, and LDHx) were evaluated in 68 infertile men and in a control of 10 fertile men. No statistically significant difference was found in the value of the 5 isoenzymes (LDH 1-5) in the different subgroups of infertile men and in the comparison of the infertile men with the control group. LDHx was detected in the infertile men, regardless of the cause of infertility, as long as the sperm concentration was greater than or equal to 16 million/ml, LDHx was not detected not only in all azoospermic men but, also, in those infertile men whose sperm count was less than or equal to 15 million/ml. There is a positive relationship (p less than 0.001) between LDHx and sperm concentration, but there is no such relationship between LDHx and % motility.