Pro: Should Varicocele Be Repaired in Azoospermic Infertile Men?

The role of varicocele repair (VR) in infertile men with non-obstructive azoospermia (NOA) and varicocele is controversial in the current guidelines, despite available studies. This study aims to assess the impact of VR on testicular sperm retrieval, sperm recovery from the ejaculate, and clinical pregnancy rates in infertile men with NOA and clinical varicocele through a systematic review and meta-analysis (SRMA) of controlled studies. A systematic literature search was conducted using the Scopus and PubMed databases up to November 2023. Among the 1,847 articles retrieved, five observational controlled studies comparing reproductive outcomes between infertile men with NOA and clinical varicocele who underwent VR, and a control group that received no treatment, met the inclusion criteria for this SRMA. The selected studies included 269 men with NOA who underwent VR before the testicular sperm extraction (TESE) procedure and 364 men who did not undergo VR. The pooled estimate demonstrated a significantly higher odds ratio (OR) of 2.17 (95% confidence interval [95% CI]: 1.17-4.01, p=0.01) for surgical sperm retrieval in the VR group. VR significantly increased the likelihood of sperm appearance in the ejaculate, with an OR of 7.8 (95% CI: 3.59-16.94, p<0.001). Besides, VR provided a significantly greater clinical pregnancy rate with intracytoplasmic sperm injection (ICSI) compared to non-operated men (OR: 2.18, 95% CI: 1.03-4.60; p=0.04). This is the first SRMA, consisting of only controlled studies, to demonstrate that VR performed prior to TESE in men with NOA significantly improves sperm production as reflected in the spontaneous appearance of sperm in the semen and higher odds of surgical sperm retrieval and clinical pregnancy compared with non-operated men. Thus, these findings highlight the potentially beneficial impact of VR in men with NOA and clinical varicocele.

The effect of varicocele repair on male fertility remains controversial. Not all men with varicoceles will experience subfertility. While most men with abnormal semen parameters will experience some degree of improvement after a varicocele repair, not all men undergoing varicocele repair will have an improvement in their fertility after varicocele repair. We performed a systematic review of the literature, looking at factors that may predict for an improvement in reproductive outcomes after varicocele repair. This would be useful to target efforts directed at these patient populations. The literature supports that men with higher-grade varicoceles will have larger improvements in their semen parameters. In addition, a higher baseline sperm concentration or total motile sperm count (TMC) is predictive of a larger post-repair TMC and also seems to correlate with pregnancy outcomes. With respect to reproductive outcomes, higher baseline sperm concentration does seem to predict for natural pregnancy or assisted reproductive technology (ART) pregnancy rates after varicocele repair. In addition, varicocele repair does seem to reduce the need for more invasive modalities of ART. In accordance with the American Urological Association (AUA) and American Society for Reproductive Medicine (ASRM) guidelines, repair of subclinical varicoceles does not seem to prognostically improve outcomes. Varicocele repair does seem to restore sperm to the ejaculate in 20–40% of men with nonobstructive azoospermia, but the factors predicting for this are still being determined. The prognostic role of hormones, patient age, body mass index, pre-repair imaging characteristics, DNA fragmentation, and histology is still being determined. The prognostic value of genetics will likely play an increasing role in the future in predicting which males will respond favorably to varicocele repair; however, data looking specifically at its prognostic role in patients undergoing varicocelectomy is still lacking at this time.

Objective. To determine the histopathological differences after varicocele repair in testicular tissue in males with nonobstructive azoospermia. Methods. Between 2009 and 2014, 45 men with complete azoospermia and palpable varicocele, presenting with primary infertility of at least 1 year, undergoing varicocele repair at our institution were selected for the study. A standard systematic testicular 6-core Tru-Cut biopsy was performed during varicocele repair. Other biopsies were obtained from each testicle of all patients at the time of microscopic sperm extraction procedure. Results. Nineteen patients were selected for the study. Testicular biopsy specimens were classified as Sertoli cell only on preoperative histopathological analysis in 14 patients. After varicocele repair, focal spermatogenesis (n = 3) and late maturation arrest (n = 2) were found in these patients. Average Johnsen score was significantly increased after varicocelectomy (P = 0.003). Motile sperm was found in one patient on postoperative semen analyses and in 10 more patients in the microscopic sperm extraction procedure. Preoperative high serum follicle stimulating hormone level and venous reflux were significantly and negatively correlated with the increase in average Johnsen score (P < 0.05). Conclusions. Our findings suggest significant improvement in testicular histology after varicocele repair.

The Effect of Prior Varicocelectomy in Patients With Nonobstructive Azoospermia on Intracytoplasmic Sperm Injection Outcomes: A Retrospective Pilot Study

Varicoceles can be a source of elevated seminal oxidative stress (OS) and sperm DNA fragmentation (SDF). However, it remains unclear whether varicocele repair (VR) could reduce these parameters. This systematic review and meta-analysis (SRMA) aims to investigate the impact of VR on SDF and seminal malondialdehyde (MDA). A literature search was performed in Scopus, PubMed, Ovid, Embase, and Cochrane databases. This SRMA included randomized controlled trials and observational studies reporting the pre- and postoperative levels of SDF and seminal OS in infertile men with clinical varicocele that underwent VR. Subgroup analyses included techniques of VR and SDF testing. The effect size was expressed as standardized mean difference (SMD). Out of 1,632 abstracts assessed for eligibility, 29 studies with 1,491 infertile men were included. The analysis showed a significant reduction in SDF after VR, compared to preoperative values (SMD -1.125, 95% confidence interval [CI] -1.410, -0.840; p<0.0001) with high inter-study heterogeneity (I²=90.965%). Reduction in SDF was evident with microsurgical technique and non-microsurgical inguinal approaches (SMD -1.014, 95% CI -1.263, -0.765; p<0.0001, and SMD -1.495, 95% CI -2.116, -0.873; p<0.0001), respectively. Reduction in SDF was significant irrespective of testing was done by sperm chromatin dispersion (SMD -2.197, 95% CI -3.187, -1.207; p<0.0001), sperm chromatin structure assay (SMD -0.857, 95% CI -1.156, -0.559; p<0.0001) or TUNEL (SMD -1.599, 95% CI -2.478, -0.719; p<0.0001). A significant decrease in seminal MDA levels was observed following VR (SMD -2.450, 95% CI -3.903 to -0.997, p=0.001) with high inter-study heterogeneity (I²=93.7%). Using pre- and post-intervention data, this SRMA indicates a significant reduction in SDF and seminal MDA levels in infertile men with clinical varicocele treated with VR. These findings may have important implications for the future management of this selected group of infertile patients.

Reply: Always important—statistical justification for pooling heterogeneous studies?

Background: Male infertility is a major health issue worldwide. Y chromosome microdeletions are well-characterized genetic causes of male infertility. The association of partial AZFc deletions (gr/gr, b2/b3, and b1/b3) with male infertility is not well confirmed in diverse populations. The purpose of the present study was to investigate the frequency of partial AZFc deletions and their association with male infertility in a population from Northwestern China. Methods: Multiplex polymerase chain reaction was used to detect partial AZFc deletions in 228 infertile patients. We analyzed 141 cases of azoospermia (AS), 87 cases of oligozoospermia (OS), and 200 fertile controls. Results: Our data showed that the frequency of a b2/b3 deletion in infertile men, men with AS, men with OS, and controls was 3.51, 2.13, 5.75, and 0.00%, respectively. The frequency of this deletion was significantly different between the infertile group and the control group (3.51 vs. 0.00%, respectively, p = 0.021) and between the OS group and the control group (5.75 vs. 0.00%, respectively, p = 0.003). The frequency of a gr/gr deletion in each group was 11.84, 9.22, 16.09, and 7.50%, respectively. The frequency of a gr/gr deletion was significantly different between the OS group and the control group (16.09 vs. 7.50%, respectively, p = 0.026) but not between the infertile group and the control group (11.84 vs. 7.50%, p = 0.132) or the AS group and the control group (9.22 vs. 7.50%, p = 0.569). The frequency of a b1/b3 deletion was 0.44, 0.71, 0.00, and 3.00%, respectively. For this deletion, there was no significant difference between the infertile (0.44 vs. 3.00%, p = 0.089), AS (0.71 vs. 3.00%, p = 0.276), and OS groups (0.00 vs. 3.00%, p = 0.236) and the control group. Conclusions: Our results suggest that the b2/b3 deletion might be associated with male infertility and that the gr/gr deletion might be associated with spermatogenic failure in men with OS in Northwestern China (Ningxia).

MP01-09 GENE DOSAGE CHANGES IN CRYPTORCHID MEN WITH NONOBSTRUCTIVE AZOOSPERMIA

To analyze the presence of potentially pathogenic variants of 29 candidate genes known to cause spermatogenic failure (SPGF) in patients with non-obstructive azoospermia (NOA) who underwent testicular histology. Forty-eight patients with unexplained NOA referred to the Department of Transfusion Medicine and Transplantation Biology, University Hospital Center Zagreb, Zagreb, Croatia for testicular biopsy. They were divided into three groups: those who had cryptorchidism (n=9), those with varicocele (n=14), and those with idiopathic NOA (n=25). All included patients underwent blood withdrawal for next-generation sequencing (NGS) analysis and gene sequencing. We found a possible genetic cause in 4 patients with idiopathic NOA (16%) and in 2 with cryptorchidism (22%). No pathogenic or possibly pathogenic mutations were identified in patients with varicocele. Variants of undetermined significance (VUS) were found in 11 patients with idiopathic NOA (44%), 3 with cryptorchidism (33%), and 8 patients with varicocele (57%). VUSs of the USP9Y gene were the most frequently as they were found in 14 out of 48 patients (29%). In particular, the VUS USP9Y c.7434+14del was found in 11 patients. They showed varied histological pictures, including Sertoli cell-only syndrome, mixed atrophy, and hypospermatogenesis, regardless of cryptorchidism or varicocele. No direct correlation was found between the gene mutation/variant and the testicular histological picture. Different mutations of the same gene cause various testicular histological pictures. These results suggest that it is not the gene itself but the type of mutation/variation that determines the testicular histology picture. Based on the data presented above, it remains challenging to design a genetic panel with prognostic value for the outcome of testicular sperm extraction in patients with NOA.

The most severe form of male infertility is idiopathic non-obstructive azoospermia (NOA), a complete sperm absence in the ejaculate. We performed exome sequencing in the Croatian infertile brothers with NOA and found a variant in DMRT1 (Doublesex and mab-3 related transcription factor 1) gene that was further assessed by the EMSA assay and molecular dynamic simulations. We additionally screened for DMRT1 mutations in 1940 infertile men diagnosed with spermatogenic failure, 644 normozoospermic controls, and 105 females with primary ovarian insufficiency (POI) recruited to the GEnetics of Male INfertility Initiative (GEMINI) or Estonian Andrology (ESTAND) cohorts. DMRT1 p.Pro74Leu (chr9:g.842059C > T) variant was detected in infertile brothers in the highly conserved position within the DNA binding DM domain of the protein. EMSA assay showed reduced DNA binding of DMRT1P74L and molecular dynamic simulations showed differences in structural and dynamical properties between the wild type protein and DMRT1P74L. Plausible disease-causing DMRT1 variants were only identified in infertile men (13/1940; 0.67%), and none in 639 fertile controls. Burden testing showed an excess of rare deleterious DM domain mutations in the infertility cohort compared to gnomAD v.4.0 population-based controls (Fisher's exact test, p = 1.44 x 10-5). Three rare deleterious variants in DMRT1 were found in 104 cases of POI. The findings of this study strengthen the evidence of DMRT1 variants being a causal factor for male infertility and provide the distribution of likely pathogenic variants across the gene. This is also the first study to suggest that DMRT1 variants may also be linked to POI.

&lt;p&gt;The outcomes of varicocele repair in non-obstructive azoospermic men remain the subject of controversy. Until now, small studies with small number of patients performed make it difficult to assess the efficacy of varicocele surgery in men with non-obstructive azoospermia. This review is performed to evaluate quality of the sperm among non-obstructive azoospermic men after varicocele repair. &lt;/p&gt;

EFFECT OF MICROSURGICAL VARICOCELECTOMY ON NONOBSTRUCTIVE AZOOSPERMIA (NOA) AND SEVERELY OLIGOZOOSPERMIA

Abnormal DNA methylation of the male germ line is proposed as a possible mechanism causing compromised spermatogenesis in some men diagnosed with idiopathic infertility. Previous studies suggested that aberrant DNA methylation of several genes is associated with disruptions in spermatogenesis. However, little information is available on DNA methylation patterns of testis-specific genes in idiopathic male infertility. To investigate the association between DAZ gene methylation patterns and spermatogenic failure, we performed an analysis of methylation patterns in 174 idiopathic infertile patients and 58 fertile controls using bisulfite-modified sequencing. We found that the methylation patterns of CpG island (CGI) in the DAZ gene promoter region were different between somatic cells and spermatic cells in the control group. DAZ gene methylation patterns among groups with different spermatogenic status were the same in somatic cells, completely methylated, and in spermatic cells. The results were concordant, except for the group with azoospermia (AZ) which were completely unmethylated. Our data indicate that the methylation patterns of the DAZ gene are not associated with spermatogenic failure. This suggests that epigenetic modification of DAZ is unlikely to be involved in the etiology of spermatogenic failure.

Etiology of azoospermia in 100 consecutive nonvasectomized men

Report on management of obstructive azoospermia