PDE11A gene polymorphism in testicular cancer: sperm parameters and hormonal profile.

PURPOSE: Testicular germ cell tumours (TGCTs) is the most common malignancy among young adult males. The etiology is multifactorial and both environmental and genetic factors play an important role in the origin and development of TGCT. Genetic susceptibility may result from the interaction of multiple common and low-penetrance genetic variants and one of the main candidate genes is PDE11A. Many PDE11A polymorphisms were found responsible for a reduced PDE activity in TGCT patients, who often also display impaired hormone and sperm profile. The aim of this study was to investigate testicular function and PDE11A sequence in testicular cancer cases. METHODS: Semen analysis was performed in 116 patients with unilateral and bilateral sporadic TGCTs and in 120 cancer-free controls. We also investigated hormone profile and PDE11A polymorphisms using peripheral blood samples. RESULTS: Our data revealed that TGCT patients showed lower testosterone levels, higher gonadotropins levels and worse semen quality than controls, although the mean and the medians of sperm parameters are within the reference limits. PDE11A sequencing detected ten polymorphisms not yet associated with TGCTs before. Among these, G223A in homozygosity and A288G in heterozygosity were significantly associated with a lower risk of testicular tumour and they displayed a positive correlation with total sperm number. CONCLUSIONS: Our findings highlight the key role of PDE11A in testis and suggest the presence of an underlying complex and fine molecular mechanism which controls testis-specific gene expression and susceptibility to testicular cancer.

In vitro effects of myo-inositol on normokinetic human semen samples with nonlinear motility.

OBJECTIVE: As myo-inositol (MI) deficiency has been associated with impaired sperm quality, we aimed to assess its effects on sperm kinetics objectively, using a computer-aided sperm analysis (CASA) system. PATIENTS AND METHODS: We evaluated 59 normokinetic semen samples with nonlinear progressive motility before and after incubation with a solution of MI. The samples were collected from healthy subjects aged 20-40 years who were attending our Laboratory of Seminology for fertility screening. RESULTS: We found a significant increase in linear progressive motility (28.2% ± 10.8 vs. 30.9% ± 11.0, T0 vs. T1 respectively; p <0.001) and a significant reduction in nonlinear progressive motility (21.0% ± 9.9 vs. 18.1% ± 10.2, T0 vs. T1 respectively; p <0.001) after incubation with MI. CASA analysis revealed a significant increase in curvilinear velocity (VCL) (65.0 ± 19.0 vs. 67.9 ± 20.4 µm/s, T0 vs. T1 respectively; p = 0.049). Overall, there was an increase in VCL in 42/59 samples (about 70%), mainly from non-smokers. CONCLUSIONS: These results suggest that MI has a positive in vitro effect on semen samples, but confirmation is needed through further studies taking into account factors capable of modulating MI response, such as smoking and obesity.

Molecular study of human sperm RNA: Ropporin and CABYR in asthenozoospermia.

BACKGROUND: Sperm motility is an essential aspect of human fertility. Sperm contain an abundance of transcripts, thought to be remnants of mRNA, which comprise a genetic fingerprint and can be considered a historic record of gene expression during spermatogenesis. The aberrant expression of numerous genes has been found to contribute to impaired sperm motility; these include ROPN1 (rhophilin associated tail protein 1), which encodes a component of the fibrous sheath of the mammalian sperm flagella, and CABYR (calcium-binding tyrosine-(Y)-phosphorylation-regulated protein), which plays an important role in calcium activation and modulation. The aim of this study was to investigate ROPN1 and CABYR gene co-expression in asthenozoospermic semen samples in comparison with normozoospermic samples. METHODS: We studied 120 semen samples (60 normozoospermic and 60 asthenozoospermic) from Caucasian patients attending our centre for an andrological check-up. Total RNA was extracted from purified spermatozoa with RNeasy mini kit. ROPN1 and CABYR mRNA expression was analysed using RT-qPCR. Continuous variables were described as means ± standard deviations. RESULTS: ROPN1 and CABYR mRNA were simultaneously downregulated in asthenozoospermic in comparison with normozoospermic samples. There was also a positive correlation between total progressive motility and ROPN1 and CABYR gene expression and between total motile sperm number and ROPN1 and CABYR gene expression. CONCLUSIONS: The results demonstrated downregulation of both ROPN1 and CABYR in asthenozoospermic samples and importantly, a positive correlation between the expression of the two genes, suggesting that ROPN1 and CABYR co-expression is a prerequisite for normal flagellar function and sperm motility.

Varicocele and semen quality: a retrospective case-control study of 4230 patients from a single centre.

PURPOSE: The aim of the present study is to assess impairment of spermatogenesis induced by varicocele in, to our knowledge, the largest single-centre caseload available to date. MATERIALS AND METHODS: We conducted a retrospective study on 4230 consecutive patients attending our Department for andrological outpatient assessment and preconception check-ups between 2011 and 2014. A total of 2113 patients had varicocele (Group V), while the remaining 2117 were selected as the control group (Group C). All patients were divided into age classes (<17, 18-28, 29-39 and ≥40 years), and Group V patients were classified as "low" (I-II) or "high" (III-IV) grade. RESULTS: Varicocele patients had a higher mean height than controls, as well as lower BMI. There was also a statistically significant reduction in the concentration/mL and the total sperm number in Group V against Group C. When stratified by age, values for all semen parameters were significantly worse in the older than in the younger age classes in both Group V and Group C, except for concentration/mL and total sperm number in the 29-39 and ≥40 age classes in both groups. A multivariable logistic regression analysis showed that factors independently predicting the presence of varicocele were older age, higher BMI and smoking for more than 10 years. CONCLUSIONS: Varicocele patients show worse semen parameters compared to controls, although their values were still within WHO reference limits. Semen quality is further worsened by increased age, grade and chronic smoking.

Androgen receptor polymorphisms and testicular cancer risk.

Testicular cancer (TC) is currently the most common malignant solid tumour in Caucasian males aged 15-39 years. Epidemiological evidence suggests that its onset may be due to an imbalance in the action of steroidal sex hormones and their receptors. A faulty androgen receptor signalling pathway can, in fact, cause various male reproductive disorders. The androgen receptor (AR) gene has two polymorphic segments consisting of CAG and GGC repeats. The length of CAG repeats has been shown to affect the regulation of AR activity. In our study, we used fragment analysis to evaluate the AR gene repeats of 302 TC patients and 322 controls, to establish if there is any association between repeat number and TC. This study of the largest Italian caseload investigated to date highlighted three particularly significant aspects. First, a CAG repeat number of ≥25 may be considered a risk factor for the onset of TC, given its greater frequency in patients in comparison with controls. This difference became significant for the non-seminoma group. Second, men with CAG repeats below 21 or above 24 were found to have a, respectively, 50 and 76% higher risk of TC than those with CAG 21-24, suggesting that these too can be considered a risk factor for TC. Finally, stage II patients were more likely to have a CAG repeat number <21 or >24 than stage I patients.