Azoospermic Men Research Articles

P-712 Transcriptomic Prediction of Spermatogenesis in the Ejaculates of Nonobstructive Azoospermic Men and Genomic Characterization of their Embryo Developmental Competence

Abstract Study question Can the genomic assessment of ejaculates from azoospermic men predict successful sperm extraction at testicular biopsy (TESE) and provide information on their reproductive potential? Summary answer RNAseq on the ejaculates of NOA men identified transcriptomic predictors of successful TESE outcome, while DNAseq evidenced gene-related mechanistics linked to reproductive performance. What is known already The most puzzling form of azoospermia is the testicular type. The chances of successful surgical sperm retrieval for these cases can be unpredictable, even when a benign histopathology has been obtained. The ability to identify individuals with complete loss of spermatogenesis at the seminiferous tubule level would help eliminate anesthesia and surgical risks, as well as financial and emotional distress. We propose to noninvasively assess the ejaculates from these men to predict successful TESE sperm retrieval. We utilize DNAseq to assess the impact of germline mutations on the embryo developmental competence of the surgically retrieved gametes. Study design, size, duration Over 4 years, we recruited men with normal peripheral karyotypes and azoospermia (n = 23), confirmed by absence of spermatozoa in extensive semen analyses. We performed epigenetic assessments on their ejaculates to compare transcriptomic profiles according to whether spermatozoa were successfully retrieved after TESE ((+)Sperm,n=11), or not ((-)Sperm,n=12). We then performed DNAseq (IRB 1006101085) on the testicular spermatozoa retrieved, to assess their ability to generate a pregnancy (fertile) or not (infertile), while controlling for maternal age. Participants/materials, setting, methods RNA was isolated from seminal fluid for epigenetic assessment (21,855 genes). A > 1 absolute log2fold change and P<0.0005 were considered significant. Transcriptomic profiles were compared between the (-)Sperm and (+)Sperm cohorts. For genetic assessment, surgically retrieved spermatozoal DNA from the (+)Sperm and obstructive azoospermic (OA) control (n = 19) cohorts were extracted and amplified. Gene mutation profiles, detected by CLC Genomic Server 9.0, were assessed for the (+)Sperm cohort according to ICSI outcomes, compared to the OA-control. Main results and the role of chance Transcriptomic analyses on seminal plasma from the 23 NOA men identified 8 imbalanced genes, involved in spermatogenesis (TMCO5B,C10orf62,SMKR1,SPZ1), sperm function (NEU1,TPTE2), and testis development (TRPC1,IGSF11-AS1). TPTE2 was almost completely expressed in 81.8%(9/11) of the (+)Sperm cohort (paternal age, 37.1±6yrs), while IGSF11-AS1 was underexpressed in all 12 (-)Sperm men (paternal age, 34.3±5yrs). Both genes are implicated in spermatogenic defects and are normally highly expressed in testis. Interestingly, NEU1, localized on the sperm acrosome and crucial for sperm capacitation, was exclusively underexpressed in (-)Sperm while overexpressed in (+)Sperm cohorts. Genomic analyses of spermatozoal DNA from the (+)Sperm cohort was compared to a matched control of OA patients (paternal age, 36.8±7yrs). Spermatozoa from the study cohort displayed mutations on genes involved in RNA transcription (POLR2L), apoptosis (AP5M1), and spermiogenic functions (AP1S2,AP1G2,APOE), while these genes were unaffected in the OA-control. When we compared sperm genetic profiles of the successful TESE cohort (maternal age, 36.8±2yrs, paternal age, 37.1±6yrs) according to ICSI outcome, we found that spermatozoa from the 8 fertile patients, all with term pregnancies, displayed consistent mild mutations on MPIG6B (stem cell lineage differentiation) while gametes from their 3 infertile counterparts carried severe mutations on genes (MBD5,CCAR1,PMEPA1,POLK,REC8,REPIN1,MAPRE3,ARL4C) involved in early embryonic development, and therefore failed to achieve embryo implantation. Limitations, reasons for caution Although we predicted, by transcriptomic analyses of ejaculates from NOA men, the presence of spermatozoa and characterized their embryo developmental competence through genetic profiling, these preliminary results need validation in a larger study cohort. Moreover, although controlled for, a confounding female factor cannot be undoubtfully excluded. Wider implications of the findings Epigenetic assessment of seminal fluid from NOA men can noninvasively identify biomarkers capable of predicting complete loss of spermatogenesis, sparing these patients from unnecessary surgery. Gene profiling can identify gametes capable of supporting term pregnancies, laying the groundwork for precision medicine in male infertility screening. Trial registration number n/a

Molecular genetic analysis of 1,980cases of male infertility.

The present study aimed to investigate the occurrence of chromosomal karyotype abnormalities and azoospermia factor (AZF) microdeletion on the long arm of the Y chromosome (Yq) in infertile men, and to determine their association with infertility to ultimately improve clinical outcomes in these patients. A total of 1,980 azoospermic and oligospermic men from the outpatient department of the Fujian Maternity and Child Health Hospital (Fuzhou, China) were recruited between January 2016 and December 2019. Peripheral blood was used for karyotype analysis; AZF microdeletion analysis of the Yq was performed using capillary electrophoresis. Among the 1,980 patients, 178 had chromosomal abnormalities (9.0%; 178/1,980), of whom 98 had an abnormal number of chromosomes. Among the abnormal karyotypes, the most common was 47, XXY (80/178; 44.9%). AZF microdeletion on the Yq occurred at a rate of 10.66% (211/1,980); the most common type was the AZFb/c deletion (sY1192; 140/211; 66.4%). The present findings showed that karyotype abnormalities and AZF gene microdeletion are important drivers of male infertility. Specifically, men with Yqh- and del(Y)(q11) had a higher risk of AZF microdeletion. These results suggested that patient treatment could be personalized based on routine molecular genetic analysis, which could further alleviate the economic and emotional burden of undergoing redundant or ineffective treatments.

Repeat Microdissection Testicular Sperm Extraction in Azoospermic Men with Nonmosaic Klinefelter Syndrome

Introduction. To investigate the predictive factors for successful repeat microdissection testicular sperm extraction attempts in patients with Klinefelter syndrome. Methods. A total of 28 azoospermic men with nonmosaic Klinefelter syndrome who have received microdissection testicular sperm extraction twice with successful initial microdissection testicular sperm extraction attempts in our institute were studied. Outcome variables (age, serum follicle-stimulating hormone, luteinizing hormone, testosterone, prolactin, and estradiol) of azoospermic men with nonmosaic Klinefelter syndrome and a successful 2nd surgical sperm retrieval attempt (group A) were compared to those with an unsuccessful 2nd sperm retrieval attempt (group B). Results. Twenty-one patients (75%) had successful sperm recovery at the 2nd microdissection testicular sperm extraction attempt. The mean testosterone level at baseline and before the 1st microdissection testicular sperm extraction attempt was higher in group A than in group B (2.7 vs. 0.9 ng/mL, p < 0.01 , and 3.9 vs. 1.1 ng/mL, p = 0.02 ). Receiver operating characteristic curve analysis identified the threshold baseline testosterone concentration (1.5 ng/mL) of patients with Klinefelter syndrome in predicting successful 2nd sperm retrieval attempts and revealed positive and negative predictive values of 94.44% and 60%, respectively. Conclusion. Azoospermic men with Klinefelter syndrome presenting with low testosterone levels and successful sperm recovery during the first microdissection testicular sperm extraction procedure are unlikely to retrieve sperm on the 2nd microdissection testicular sperm extraction attempt. Hence, these patients should be properly counseled before sperm retrieval.

Generation of Haploid Spermatids on Silk Fibroin-Alginate-Laminin-Based Porous 3D Scaffolds.

In vitro production of sperm is a desirable idea for fertility preservation in azoospermic men and prepubertal boys suffering from cancer. In this study, a biocompatible porous scaffold based on a triad mixture of silk fibroin (SF), alginate (Alg), and laminin (LM) is developed to facilitate the differentiation of mouse spermatogonia stem cells (SSCs). Following SF extraction, the content is analyzed by SDS-PAGE and stable porous 3D scaffolds are successfully prepared by merely Alg, SF, and a combination of Alg-SF, or Alg-SF-LM through freeze-drying. Then, the biomimetic scaffolds are characterized regarding the structural and biological properties, water absorption capacity, biocompatibility, biodegradability, and mechanical behavior. Neonatal mice testicular cells are seeded on three-dimensional scaffolds and their differentiation efficiency is evaluated using real-time PCR, flow cytometry, immunohistochemistry. Blend matrices showed uniform porous microstructures with interconnected networks, which maintained long-term stability and mechanical properties better than homogenous structures. Molecular analysis of the cells after 21 days of culture showed that the expression of differentiation-related proteins in cells that are developed in composite scaffolds is significantly higher than in other groups. The application of a composite system can lead to the differentiation of SSCs, paving the way for a novel infertility treatment landscape in the future.

Preservation of fertility in patients with testicular germ cell tumors: oncological, legal, and religious aspects

Survival rates for cancer patients continue to steadily increase due to improvement of effectiveness of current treatments. However, despite significant oncological results, one should not forget about the quality of life of this cohort of patients, in particular those who undergo gonadotoxic cancer therapy with development of premature ovarian failure in women and azoospermia in men. Preservation of fertility in both women and men with cancer is currently possible and should be integrated at all levels of cancer care. In this regard, the main purpose of this review is to consider the topic of fertility in men treated for cancer in the context of various aspects of human life.

Prolactin and spermatogenesis: new lights on the interplay between prolactin and sperm parameters.

To clarify the relationship between one the most gender-specific hormone, i.e. prolactin (PRL), and semen parameters in men. A retrospective, observational, cohort, real-world study was carried out, enrolling all men performing a semen analysis and PRL examination from 2010 to 2022. For each patient, the first semen analys was extracted, associated to PRL, total testosterone (TT), follicle stimulating hormone (FSH) and luteinizing hormone (LH). Hyperprolactinaemia (>35 ng/mL) was excluded. 1211 subjects were included. PRL serum levels were lower in normozoospermia compared to azoospermia (p = 0.002) and altered semen parameters (p = 0.048) groups. TT serum levels were not different among groups (p = 0.122). Excluding azoospermic men, PRL serum levels were lower in normozoospermic patients, when compared to other groups of semen alterations. An inverse correlation was detected between PRL and sperm concentration. Considering normozospermic subjects, PRL was directly related to both non-progressive sperm motility (p = 0.014) and normal sperm morphology (p = 0.040). Subdiving the cohort in quartiles according to PRL distribution, the highest motilities were observed in the second PRL quartile (8.30-11.10 ng/mL) and asthenozoospermia was significantly predicted by FSH (p < 0.001) and second PRL quartile (p = 0.045). The PRL-spermatogenesis connection seems to be mild, although low-normal PRL levels are associated with the best spermatogenetic profile. PRL serum levels could mirror the immunoregulatory status within the testis, suggesting that there is a sort of 'PRL optimal window' reflecting an efficent spermatogenesis. Alternatively, men with good semen parameters might have a higher central dopaminergic tone resulting in low PRL levels.

Unilateral Versus Bilateral Vasoepididymal Anastomosis for Idiopathic Obstructive Azoospermia: A Randomised Controlled Trial

BackgroundMen with idiopathic obstructive azoospermia (OA) are candidates for surgical reconstruction with a vasoepididymal anastomosis (VEA) performed on one or both testis. There are no randomised trials comparing the success of unilateral versus bilateral VEA. ObjectiveWe conducted a randomised trial to compare the two surgical options. Design, setting, and participantsBetween April 2017 and March 2022, men with infertility due to idiopathic OA were randomised to a unilateral (group 1) or bilateral (group 2) VEA in an ethics committee–approved clinical trial, registered with the Clinical Trials Registry. Outcome measurements and statistical analysisThe primary outcome was successful surgery, defined as appearance of sperm in the ejaculate, evaluated at 3 mo intervals after surgery. Additional outcomes were pregnancy rates and complications between the two groups. Men with successful surgery were compared with those without patency to identify the predictors of success. Results and limitationsFifty-four men fulfilled the criteria and 52 who completed follow-up were included in the analysis. The overall patency rate was 36.5% (19/52 individuals). This was higher in men with bilateral surgery (12/26 patients, 46%) than in those with unilateral surgery (7/26 patients, 27%) but was not statistically significant (p = 0.1). The overall pregnancy rate with ejaculated sperm was significantly higher in the bilateral surgery group (4 vs 0, p = 0.037), while the spontaneous conception rate was higher but not statistically significant (3 vs 0, p = 0.074). The complication rates in the two groups were similar (p = 0.7), and all complications were Clavien-Dindo grade 1. Although bilateral surgery and presence of sperm in epididymal fluid were higher in men with patency, these were not statistically significant. ConclusionsA bilateral VEA was associated with higher patency and spontaneous pregnancy rates than unilateral surgery, but the results were not statistically significant. However, the overall pregnancy rate with ejaculated sperm, spontaneous and assisted, was significantly higher in the bilateral surgery group. Patient summaryIn this study, we compared between unilateral and bilateral reconstructive surgery in azoospermic men and found better overall success with bilateral surgery. However, these results were not statistically significant.

Anti-Mullerian Hormone Is a Significant Marker for Male Infertility

To evaluate serum anti-Mullerian hormone (AMH) in men with normal, reduced sperm concentration and in azoospermic men as a possible clinical marker of male factor infertility. Prospective study Private clinic of Dr. Yasir Al-Wattar and Al-Batool Teaching Hospital in Mosul. This study was conducted on one hundred male subjects aged less than 50 years for the period from July to November 2011, infertile men were classified according to their sperm count into oligospermics (n=28) and azoospermics(n=47), twenty five men were normal (n=25).Serum concentrations of FSH and Testosterone were measured using ELISA, serum Anti-Mullerian hormone concentrations were measured using AMH/MIS enzyme linked immunosorbant assay kit. Significant differences in serum AMH, FSH and Testosterone concentrations were found between normal, oligospermic &amp; azoospermic men. AMH negatively correlated with FSH in oligospermic and azoospermic men and positively with testosterone in azoospermics and negatively with testosterone in oligospermics. Anti-Mullerian hormone should be evaluated in patients with oligospermia and azoospermia, it may serve as a marker of male infertility.

New Insights into the Role of INSL-3 in the Development of Cryptorchidism

Cryptorchidism, defined as the failure of at least one or both testicles to descend into the scrotal pouches, is the most frequent (1.6–9% at birth, 1/20 males at birth) congenital anomaly encountered in newborn males, resulting in one of the most frequent causes of non-obstructive azoospermia in men. Similar to other congenital malformations, cryptorchidism is thought to be caused by endocrine and genetic factors, combined with maternal and environmental influences. The etiology of cryptorchidism is unknown, as it involves complex mechanisms aiming to control the testicular development and descent from their initial intra-abdominal location in scrotal pouches. The implication of insulin-like 3 (INSL-3) associated with its receptor (LGR8) is critical. Genetic analysis discloses functionally deleterious mutations in INSL3 and GREAT/LGR8 genes. In this literature review, we discuss and analyze the implication of INSL3 and the INSL3/LGR8 mutation in the occurrence of cryptorchidism in both human and animal models.

Testicular Apoptosis and Proliferation in Relation to Body Mass Index and m-TESE Success in Nonobstructive Azoospermic Men

Male infertility occurs due to the disruption of the balance of proliferation and apoptosis in the testicular tissue. It is important to show the effect of increased body mass index, which is one of the factors that disrupts this balance of apoptosis and proliferation, not only at the blood hormone level but also at the tissue level. For this reason, the present study is aimed at demonstrating the relation between body mass index and cell turnover in the testis using immunohistochemical methods. In the present study, patients were grouped as normal, overweight and obese, and as m-TESE positive and negative. The sperm retrieval rate with microsurgical testicular sperm extraction (m-TESE) was 41.67%. Histological diagnosis of testicular tissues was made with hematoxylin and eosin and Masson’s trichrome staining. Apoptosis and proliferation in the testicular tissue were demonstrated by TUNEL and PCNA immunohistochemical methods, respectively. It was concluded that BMI had no significant effect on reproductive hormone profile (FSH, LH, and testosterone), m-TESE success, apoptosis, and proliferation in testicular tissue in nonobstructive azoospermic men. In addition TUNEL positivity and proliferative index was found to be significantly correlated with testicular histology and m-TESE outcome.

MP30-08 EVALUATING THE CHANGES IN SEMEN PARAMETERS OVER TIME FOR MEN WITH AZOOSPERMIA FACTOR C (AZFc) MICRODELETIONS

You have accessJournal of UrologyCME1 Apr 2023MP30-08 EVALUATING THE CHANGES IN SEMEN PARAMETERS OVER TIME FOR MEN WITH AZOOSPERMIA FACTOR C (AZFc) MICRODELETIONS Jessica Marinaro, Jonathan Gal, Anna Mielnik, and Peter Schlegel Jessica MarinaroJessica Marinaro More articles by this author , Jonathan GalJonathan Gal More articles by this author , Anna MielnikAnna Mielnik More articles by this author , and Peter SchlegelPeter Schlegel More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003258.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Men with complete azoospermia factor C (AZFc) microdeletions may have variable degrees of spermatogenic failure. While some are persistently azoospermic, most intermittently or consistently have sperm in the ejaculate. It has been proposed that these men may experience a temporal decline in semen quality with age. The objective of this study was to assess changes in semen parameters over time among men with complete AZFc deletions. METHODS: We retrospectively reviewed 163 men with complete AZFc deletions who were tested at our center from 4/1996 to 7/2022. Of these, 41 had ≥2 semen analyses performed (SA). ANOVA tests were used to compare mean demographic values between azoospermic, intermittently azoospermic, and non-azoospermic men. Wilcoxon matched-pairs signed rank tests were used to compare initial and final semen analyses. RESULTS: In total, 179 SAs were reviewed. The mean number of SAs per patient was 4.4 ± 2.2. The mean time between each SA was 189.6 ± 420.0 days, and the mean time between the first and last SA was 638.9 ± 850.9 days. Of 41 men, 13 (32%) were azoospermic, 9 (22%) intermittently azoospermic, and 19 (46%) were non-azoospermic. Of the non-azoospermic men, most (n=14, 73.7%) had severe oligospermia (<1 million/mL).Of 9 men with intermittent azoospermia, most had an increase in sperm concentration (n=8, 89%), total sperm count (TSC) (n=8, 89%), and total motile sperm count (TMSC) (n=6, 67%) between the first and last SA. Similarly, most non-azoospermic men also had an increase in sperm concentration (n=12, 63%), TSC (n=11, 58%), and TMSC (n=11, 58%). In comparing semen parameters, men with intermittent azoospermia had an increase in sperm concentration (p=0.0078) and TSC (p=0.0078) between the first and last SA, but no change in TMSC. There was no difference in concentration, TSC, or TMSC over time for non-azoospermic men.While non-azoospermic men were older (mean: 37.0 +/- 5.6 years; p=0.004) versus intermittently azoospermic (mean: 33.4 +/- 3.3 years) or azoospermic men (mean: 30.3 +/- 5.6 years), there were no differences in baseline testosterone (p=0.24), baseline FSH (p=0.31), or testis size (p=0.17 right, p=0.50 left). CONCLUSIONS: In this study, there was no significant decline in sperm concentration, TSC, or TMSC over time for men with complete AZFc deletions; rather, sperm number tended to increase over time. These data suggest that men with AZFc deletions are not at substantive risk of decline in sperm production and do not need to take aggressive early measures for fertility preservation. Source of Funding: Authors JM and JG are supported in part by the Frederick J. and Theresa Dow Wallace Fund of the New York Community Trust. © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e393 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jessica Marinaro More articles by this author Jonathan Gal More articles by this author Anna Mielnik More articles by this author Peter Schlegel More articles by this author Expand All Advertisement PDF downloadLoading ...

PD18-06 PREDICTORS OF AZOOSPERMIA IN MEN WITH TESTIS CANCER

You have accessJournal of UrologyCME1 Apr 2023PD18-06 PREDICTORS OF AZOOSPERMIA IN MEN WITH TESTIS CANCER Luis F. Novaes, Richard S. Matulewicz, Joel Sheinfeld, John P. Mulhall, and Jose M. Flores Luis F. NovaesLuis F. Novaes More articles by this author , Richard S. MatulewiczRichard S. Matulewicz More articles by this author , Joel SheinfeldJoel Sheinfeld More articles by this author , John P. MulhallJohn P. Mulhall More articles by this author , and Jose M. FloresJose M. Flores More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003273.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Testis cancer (TC) is the most common cancer in young men. Many men with TC are also found to be sub-fertile at the time of diagnosis. Standard treatment modalities often negatively affect reproductive potential, making fertility a vital consideration in this population. We aimed to evaluate predictors of azoospermia in the pre-radical orchiectomy (RO) semen analysis (SA) in men with TC to inform patient counseling and guide fertility preservation. METHODS: Our analytic cohort included (i) men treated with RO for testicular cancer at our institution from January 2010 to August 2022 who had a (ii) SA, and (iii) fertility endocrine profile available prior to surgery. SA evaluated sperm concentration categorized as: azoospermia (0 M/ml), severe oligospermia (<5 M/ml), oligospermia (5-15 M/ml), normospermia (>15 M/ml). Parameters were reported as frequencies or median (IQR). Patient demographics, total testosterone (T), FSH levels, and testis ultrasound (TUS) parameters were also collected. TUS parameters analyzed were testis volume of each testis, tumor volume, number of tumors, and presence of microcalcifications. Multivariable analysis (MVA) was used to explore factors associated with azoospermia. Factors included in the model were testis volume of the side affected by tumor, tumor volume, FSH levels, and the presence of microcalcifications. RESULTS: A total of 97 men met inclusion criteria. Median age was 30 (25, 35) years. Median SA volume was 3 (1.5, 4) mL, median concentration 13 (0.3, 40) M/ml; 15% had azoospermia, 20% severe oligospermia, 19% oligospermia, 46% normospermia. Median total T was 393 (319, 529) ng/dL; 19% had T level ≤300 ng/dL. Median FSH was 18 (3, 32) IU/mL. The median testis volume was 15 (10, 23) cc, the median tumor volume was 4 (1, 18) cc. 96% of tumors were unilateral, and 29% had microcalcifications. In men with testis volume of the side affected by the tumor ≤10cc the rate of azoospermia was 21%, versus 13% in men with testis volume >10 cc. On MVA, testis volume of the side affected by the tumor was the only factor statistically associated with azoospermia. CONCLUSIONS: For men with testis cancer, lower testis volume of the side affected by the tumor is independently associated with azoospermia. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e504 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Luis F. Novaes More articles by this author Richard S. Matulewicz More articles by this author Joel Sheinfeld More articles by this author John P. Mulhall More articles by this author Jose M. Flores More articles by this author Expand All Advertisement PDF downloadLoading ...

PD18-02 DESCRIBING PATTERNS OF FAMILIAL CANCER RISK IN SUBFERTILE MEN USING POPULATION PEDIGREE DATA

You have accessJournal of UrologyCME1 Apr 2023PD18-02 DESCRIBING PATTERNS OF FAMILIAL CANCER RISK IN SUBFERTILE MEN USING POPULATION PEDIGREE DATA Joemy Ramsay, Joshua Horns, Benjamin Emery, Kenneth Aston, and James Hotaling Joemy RamsayJoemy Ramsay More articles by this author , Joshua HornsJoshua Horns More articles by this author , Benjamin EmeryBenjamin Emery More articles by this author , Kenneth AstonKenneth Aston More articles by this author , and James HotalingJames Hotaling More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003273.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Subfertile men and their relatives show increased risk for certain cancers including testis, pediatric leukemia/lymphoma, thyroid, and prostate. We assessed risk for multiple cancers simultaneously to identify distinct familial multicancer patterns (FMPs) in azoospermic and oligozoospermic men. METHODS: We identified azoospermic (N=809) and oligozoospermic (N=669) men in the Subfertility, Health and Assisted Reproduction (SHARE) cohort and their family members out to third-degree relatives (azoo: N=27,148; oligo: N=23,626). Cancer cases were identified using the Utah Cancer Registry, a population-based SEER registry with statewide cancer records from 1966. Families with ≥10 individuals who lived in Utah for ≥1 year 1966-2017 were included in the analysis (azoo: N=426 families, 21,361 individuals; oligo: N=360 families, 18,818 individuals). The famcluster R package was used to cluster families on familial standardized incidence ratios (FSIRs). FSIRs were calculated across 34 cancer types and a weighted Gower distance with k-mediods clustering (k=2:20) was used to identify FMPs. k was selected using silhouette plots and consensus clusters with 1,000 bootstrap replicates. FMPs were assessed separately for azoospermia and oligospermia families. RESULTS: 13 FMPs were identified in azoospermia families and 12 FMPs in oligozoospermia families. Distinct overall patterns of familial cancer were observed between azoospermia and oligozoospermia families (Table 1). 12 out of 13 FMPs (36% of families) showed significantly increased risk for ≥1 cancer in azoospermia families and 11 out of 12 FMPs (40% of the families) showed significantly increased cancer risk in oligozoospermia families (Figure 1). CONCLUSIONS: This analysis described FMPs for subfertile men, with 36-40% of families showing elevated risk for ≥1 cancer. FMPs were unique between azoospermia and oligozoospermia families, suggesting heterogeneity in familial cancer risk by both type of subfertility and within subfertility type. Source of Funding: GEMS: Genomic approach to connecting Elevated germline Mutation rates with male infertility and Somatic health NICHD: R01 HD106112 © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e502 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Joemy Ramsay More articles by this author Joshua Horns More articles by this author Benjamin Emery More articles by this author Kenneth Aston More articles by this author James Hotaling More articles by this author Expand All Advertisement PDF downloadLoading ...

The effect of azoospermia factor microdeletions on intracytoplasmic sperm injection results in azoospermia patients.

Y chromosome abnormalities are common in male patients with severe oligo-azoospermia. In studies with karyotype analysis and cytogenetic methods, the importance of the Y chromosome in spermatogenesis has been well understood. Deletions in the azoospermia factor (AZF) localized at the distal end of the Y chromosome adversely affect the spermatogenesis process. Our objective was to determine the frequency of AZF microdeletion in azoospermia patients who underwent microTESE. In this retrospective cohort study, 806 azoospermic men attending the In Vitro Fertilization (IVF) Center for infertility treatment between 2010 and 2022 were included. AZF deletion screening was conducted in all patients included in the study. Azoospermic patients with and without Y microdeletion were matched with the female's age, cause of infertility, number of oocytes retrieved and number of metaphase II (MII) oocytes produced and compared. The primary outcome was the live birth rate (LBR). Pregnancy rate (PR) and clinical pregnancy rates (CPR) were secondary outcomes. We detected Y microdeletion in 55 (6.82%) of 806 infertile azoospermic men and 35 of them included in the study. Although the required gonadotropin dose and the total number of retrieved oocytes were similar, clinical pregnancy rates and live birth rates were found to be significantly lower in the microdeletion patient group (21.6% vs. 43%, p<0.05; and 18.9% vs. 36%, p<0.05, respectively). Poor sperm quality in AZF microdeletion patients complicates the selection of appropriate sperm for ICSI. Therefore, it leads to a decrease in embryonic development, fertilization and pregnancy results. In order to select the best sperm for the use in ICSI procedure in this patient population, intracytoplasmic morphologically selected sperm injection (IMSI) method can be preferred to improve the cycle outcomes.

Touch print smear of testicular tissue with thionine stain for intraoperative diagnosis in azoospermic men.

To compare the diagnostic value of testicular tissue touch print smear (TPS) conducted on azoospermic patients with results obtained from histopathology and in vitro fertility (IVF) lab findings. Microdissection testicular sperm extraction was performed on a group of 148 azoospermic patients and testicular samples obtained intraoperatively. Using TPS, the samples were smeared onto a sterile slide, followed with staining using thionine. The testis tissue bulk samples were also transferred to the IVF lab, and determinations of sperm presence or absence obtained from IVF lab tests were compared with the TPS sample results. Needle testis biopsy was separately performed on a group of 360 azoospermic patients, and results of pathohistology review on the biopsies were further compared with determinations of spermatogenesis stage obtained from TPS for those patients. When compared with IVF lab results, TPS was found to have 100% (126/126) positive predictive value and 95.5% (25/26) negative predictive value for predicting sperm presence or absence, respectively. Furthermore, TPS was further found to have a 93.6% correlation (337 of 360 biopsies) with results of histological diagnoses performed by needle biopsy. Results from histology and TPS for the detection of sperm presence were concordant in 96.1% (346/360) of biopsies. Diagnosis of SCO by TPS shows the highest correlation with histopathology (98.6%), followed by complete spermatogenesis (97.5%), early maturation arrest (78.9%), and late maturation arrest (27.3%). The results support the continued use of TPS in testicular tissue analysis for more rapid assessment of spermatogenesis and for detection of spermatozoa in azoospermic subjects.

Testicular impairment in Primary Adrenal Insufficiency caused by Nicotinamide Nucleotide Transhydrogenase (NNT) deficiency - a case report: implication of oxidative stress and importance of fertility preservation

IntroductionNicotinamide nucleotide transhydrogenase (NNT) gene deficiency has recently been shown to be involved in Primary Adrenal Insufficiency (PAI). NNT encodes an inner mitochondrial membrane protein that produces large amounts of NADPH. NADPH is used in several biosynthesis pathways and the oxidoreduction of free radicals by the glutathione and thioredoxin systems in mitochondria. Patients with PAI due to NNT deficiency may also exhibit extra-adrenal manifestations, usually including gonadal impairment.Case reportWe present the case of a 35-year-old patient referred to our center for primary infertility with non-obstructive azoospermia, in a context of PAI and obesity. PAI genetic exploration carried out at the age of thirty revealed NNT deficiency due to the presence of two deleterious mutations (one on each allele) in the NNT gene. Scrotal ultrasound revealed a right Testicular Adrenal Rest Tumor (TART). Intensification of glucocorticoid therapy over the course of 8 months failed to reduce the TART volume or improve sperm production and endocrine function. No spermatozoa were found after surgical exploration of both testes, and subsequent histopathological analysis revealed bilateral Sertoli cell-only syndrome. A retrospective review of the hypothalamic-pituitary-gonadic axis hormonal assessment over 20 years showed progressive impairment of testicular function, accelerated during adulthood, leading to hypergonadotropic hypogonadism and non-obstructive azoospermia when the patient reached his thirties, while the PAI remained controlled over the same period.ConclusionThis case report provides, for the first time, direct evidence of complete germ line loss in an azoospermic man with NNT deficiency. Additional data further support the hypothesis of a determinant role of oxidative cellular damage due to reactive oxygen species (ROS) imbalance in the severe gonadal impairment observed in this NNT-deficient patient. Early and regular evaluation of gonadal function should be performed in patients with PAI, especially with NNT deficiency, as soon as the patients reach puberty. Fertility preservation options should then be provided in early adulthood for these patients.

The MAEL expression in mitochondria of human spermatozoa and the association with asthenozoospermia.

The maelstrom spermatogenic transposon silencer (MAEL) function in postmeiotic germ cells remains unclear, and its protein localization in human testis and spermatozoa awaits determination. This study aims to clarify the MAEL expression in human spermatogenesis and to explore its role in sperm function. Twenty-seven asthenozoospermic men, 40 normozoospermic controls, and three obstructive azoospermic men were enrolled. The transcripts of MAEL in the seminiferous epithelium and MAEL downstream targets were identified by bioinformatics analysis. MAEL protein expression in human testis and ejaculated sperms were examined by immunohistochemical and immunogold staining, respectively. The roles of MAEL in mitochondria function were investigated by siRNA knockdown in human H358 cells. The association between MAEL protein levels and clinical sperm features was evaluated. Abundant MAEL was expressed in spermatid and spermatozoa of the human testis. Remarkably, MAEL was located in the mitochondria of ejaculated sperm, and bioinformatics analysis identified GPX4 and UBL4B as MAEL's downstream targets. Knockdown of MAEL sabotaged mitochondria function and reduced adenosine triphosphate (ATP) production in H358 cells. MAEL, GPX4, and UBL4B expression levels were significantly decreased in asthenozoospermic sperms than in controls. The MAEL protein levels were positively correlated with GPX4 and UBL4B in human sperm. Total motile sperm count (TMSC) was positively correlated with protein levels of MAEL, GPX4, and UBL4B in ejaculated sperms. We highlight prominent MAEL expression in the intratesticular spermatid and the mitochondria of ejaculated spermatozoa. MAEL directly binds to GPX4 and UBL4B, and loss of MAEL induces mitochondrial dysfunction. MAEL-mitochondrial function-motility relationship might advance our understanding of the causes of asthenozoospermia.

Chromosomal Abnormalities in Infertile Greek Men: A Single Institution’s Experience

Chromosomal abnormalities represent a significant genetic cause of male infertility because they impair spermatogenesis. The objective of the current study was to determine the prevalence and distribution of chromosomal abnormalities in Greek men with infertility. Four hundred eighty-eight infertile men (27 azoospermic, 168 with oligospermia -98 mild, 57 moderate, 13 severe- and 293 with normospermia) undergoing In &lt;em&gt;Vitro &lt;/em&gt;Fertilization (IVF) between 2016-2022 were enrolled in the study. Thirty-eight fertile men were also studied. Chromosomal analysis of peripheral blood lymphocytes was performed using standard cytogenetic techniques. 21/488 (4.3%) of men tested had an abnormal karyotype; 13 (2.7%) had sex chromosome abnormalities and 8 (1.6%) had autosomal ones. No chromosomal aberration was detected in the control group. The frequency of chromosomal alterations was significantly higher in azoospermic men than in men with oligospermia and normospermia (37% vs 4.2% and 1.4% respectively, p &lt; 0.05). Moreover, in men with oligospermia, cytogenetic abnormalities were more common in the severe group (7.7%) followed by the moderate (5.25%) and the mild group (3%). The results of the study are by the literature. Karyotyping is suggestive especially in oligospermic /azoospermic men and before proceeding to IVF. The advent of high throughput sequencing technologies and genome-wide association studies will contribute to discovering novel promising genetic factors involved in male infertility.

MicroRNAs expression in semen and testis of azoospermic men.

MicroRNAs are involved in the regulation of spermatogenesis, are detected in semen and may be useful as molecular markers for predicting residual complete spermatogenesis in azoospermic men. To study the biomarker potential of microRNAs that are detected in semen and testicular tissue. MicroRNA profiles were analyzed in semen fractions of normozoospermic (n=3) and azoospermic (n=6) men by small RNA deep sequencing. Specific microRNAs were further analyzed by reverse transcription and quantitative polymerase chain reaction in eight testicular samples and 46 semen supernatants. The semen supernatant samples included 18 normozoospermic and 28 azoospermic men with various pathologies. The sequenced microRNA profiles of semen supernatant fraction samples were distinct from the other fractions. Significant expression differences were observed between the semen supernatant of normozoospermic and azoospermic men. Further analysis by reverse transcription and quantitative polymerase chain reaction revealed that expression of miR-202-3p was considerably reduced (undetectable in most samples) in the azoospermic semen supernatants. The expression of miR-202-3p was significantly lower in the azoospermic specimens than in the normozoospermic specimens and a trend was observed for miR-629-5p (p=0.03 and 0.06, respectively). Differences in expression levels in the semen supernatant were observed among the various pathologies but not to a level of significance, possibly because of the small subgroups. miRNA-370-3p was significantly higher in semen supernatant samples from azoospermic men without sperm cells in testis (p=0.05). In testes, the three microRNAs were expressed at higher levels in the obstructive and spermatocyte maturation arrest pathologies than in mixed atrophy and Sertoli cell only. miR-202-3p was detected in all testicular samples. MicroRNA expression profiles in semen were distinguishable between azoospermic and normozoospermic men. The microRNA profile also diverged among azoospermic men subdivided according to their testicular pathologies. The levels of specific microRNAs in testis and in the semen supernatant were not directly correlated.

Does Clomiphene citrate administration increase success rate of micro TESE in non-obstructive azoospermic men? A randomized controlled trial

Does Clomiphene citrate administration increase success rate of micro TESE in non-obstructive azoospermic men? A randomized controlled trial