Prediction of Posttreatment Spermatogenesis in Patients With Testicular Cancer by Flow Cytometric Sperm Chromatin Structure Assay (original) (raw)

Sperm integrity pre- and post-chemotherapy in men with testicular germ cell cancer

Human Reproduction, 2006

BACKGROUND: While (partial) recovery of spermatogenesis, observed by means of standard semen analysis, has been seen in testicular cancer patients after chemotherapy with cisplatin, sperm genomic integrity and its implication for the patient's fertility are poorly understood. METHODS: Semen and serum from 22 patients treated for testicular cancer were analysed pre-and post-chemotherapy. Besides routine semen analysis, sperm samples were evaluated by computerized karyometric image analysis (CKIA), chromomycin-A3 assay (CMA3 , chromatin condensation) and TdT-mediated dUTP nick-end labelling assay (TUNEL, DNA damage). Serum FSH, LH and testosterone concentrations were measured. RESULTS: Ejaculate volume decreased post-chemotherapy (P < 0.05). External sperm characteristics (CKIA morphometry) and sperm counts did not deteriorate after chemotherapy. An improvement in DNA condensation was assessed after chemotherapy (37 versus 50% and 47.5 versus 63.7% for CMA3 and CKIA respectively; both P<0.005); yet a high percentage of TUNEL-positive sperm was found in the samples (21 versus 25% for pre-and post-chemotherapy samples respectively). These values were significantly higher than those of a convenience sample of normozoospermic males attending pre-IVF screening. Serum FSH and LH (IU/l) increased after chemotherapy compared with pretreatment levels (8.1 versus 16.7 and 4.5 vs 6.8; both P < 0.05, respectively). CONCLUSIONS: Despite the improvement in sperm chromatin packaging after chemotherapy, an abnormally high percentage of DNA-damaged sperm was found in these samples. As sperm quality does not reach normal levels after treatment, it remains difficult to outline the best strategy and guidance concerning fertility potential of testicular cancer patients.

Effect of chemo- or radiotherapy on sperm parameters of testicular cancer patients

Human Reproduction, 2006

BACKGROUND: The aims of our study were to investigate the short-and long-term effects of chemo-or radiotherapy on spermatogenesis in patients with testicular cancer and to establish any correlation between pre-therapy sperm parameters, histotype and treatment type/intensity and the progress of spermatogenesis during the post-therapy period. METHODS: We evaluated 166 patients affected by testicular cancer, who cryobanked about 1 month after the removal of the cancerous testis and before beginning chemo-(CH group; n = 71) or radiotherapy (RT group; n = 95). RESULTS: For the CH group, there was a statistically significant decrease in sperm parameters, which was most significant 3 months after the end of chemotherapy. For the RT group, this decrease was most relevant 6 months after the end of radiotherapy. Two years after therapy, 3% of the CH group and 6% of the RT group remained azoospermic. To evaluate whether spermatogenesis recovery is a function of baseline semen quality, we divided each group into two subgroups by pre-therapy total sperm count (A, <40 ´ 10 6 /ejaculate; B, ³40 ´ 10 6 /ejaculate). At t 24 , subgroup A of both the CH and RT groups showed improved sperm parameters over the baseline, whereas subgroup B for both CH and RT groups showed a return of sperm parameters to those of baseline values. CONCLUSIONS: In conclusion, the recovery of spermatogenesis after chemo-or radiotherapy in our group of testicular cancer patients was not a function of pre-therapy sperm parameter quality. Cryopreservation of sperm before performing such therapy is therefore imperative.

Absence of chromosomal instability in spermatozoa of men affected by testicular cancer

Human Reproduction, 1999

Since cancer treatment using antineoplasic drugs and ionizing radiation has a negative effect on the function of the gonads, testicular cancer patients are offered the opportunity to cryopreserve their semen samples before the beginning of therapy. For this reason it would be of interest to know whether there is chromosome instability in their spermatozoa prior to any treatment. Using the interspecific human-hamster fertilization system, we have analysed a total of 340 chromosome complements from spermatozoa of control donors and 320 chromosome complements from testicular cancer patients. There were no significant differences in the frequencies of chromosomal aberrations between controls and cancer patients (9.7 and 10.3% respectively; P ϭ 0.4921). Our results indicate that spermatozoa from untreated testicular cancer patients do not show an increased chromosomal instability as compared to control donors.

Effects of the Chemotherapy Cocktail Used to Treat Testicular Cancer on Sperm Chromatin Integrity

Journal of Andrology, 2006

The incidence of testicular cancer has increased dramatically over the past 50 years. Advances in treatment, which include the coadministration of bleomycin, etoposide, and cisplatinum (BEP), have brought the cure rate to over 90%. After treatment, most patients go through a temporary period of azoo/ oligozoospermia. Although the sperm concentration in approximately 80% of the patients returns to at least 10 million/mL, little is known about the integrity of the chromatin of their germ cells. Using an animal model, we assessed DNA integrity in the spermatozoa of male rats treated for 3, 6 or 9 weeks with BEP at doses, adjusted for surface area, equivalent to 0X, 1/3X, 2/3X, or 1X of the human dose. We did not observe any difference in protamination content, as assessed by the chromomycin A3 (CMA3) assay. After 9 weeks of 1X treatment, the susceptibility of DNA to denaturation evaluated by the sperm chromatin structure assay (SCSAH)/acridine orange assay (AO) was increased, as well as the number of single and double DNA strand breaks measured by the TUNEL and COMET assays. Parameters obtained from the AO and TUNEL assays were highly correlated with the motility of the spermatozoa, suggesting that conventional sperm analysis parameters can serve as a good indicator of chromatin integrity and vice versa. Correlation studies also suggested that the parameters obtained with the different assays do not overlap, but complement each other. Thus, BEP treatment altered spermatozoal chromatin quality, and these alterations may impact adversely on progeny outcome.

Characterization of sperm chromatin quality in testicular cancer and Hodgkin's lymphoma patients prior to chemotherapy

Human Reproduction, 2008

BACKGROUND: Although the incidences of testicular cancer and Hodgkin's lymphoma have increased in young men over the past decade, combination chemotherapy has improved survival. As fertility is of importance to these patients, characterization of sperm chromatin structure is needed. We assessed sperm chromatin in testicular cancer and Hodgkin's lymphoma patients prior to chemotherapy, in comparison with control community and idiopathic infertile volunteers. METHODS: DNA damage was assessed with the sperm chromatin structure assay (SCSA), terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and comet assays; reactive thiols (SH) and DNA compaction were determined with the monobromobimane (mBBr) and chromomycin A3 (CMA3) assays, respectively. RESULTS: Both testicular cancer (37%) and Hodgkin's lymphoma (81%) patients had normospermic samples with increased DNA damage, compared with controls. Cancer patients also had higher reactive thiols and CMA3 staining, indicating low DNA compaction. CONCLUSIONS: Sperm DNA integrity and compaction were affected in testicular cancer and Hodgkin's lymphoma patients prior to chemotherapy. Although SCSA, TUNEL and comet assays all detected DNA damage, the latter was optimal for use in cancer patients. A combination of the comet assay with tests that evaluate sperm DNA compaction, such as flow cytometry-based CMA3 and mBBr assays, is a reliable strategy to characterize sperm chromatin quality in cancer patients at the time of sperm banking.

Testicular cancer and sperm DNA damage: short- and long-term effects of antineoplastic treatment

Andrology, 2014

The aim of this study was to investigate sperm DNA damage induced by chemo-and radiotherapy in patients with testicular cancer to provide data on the extent and persistence of nuclear damage that might affect individual reproductive potential. We evaluated pre-and post-antineoplastic treatment sperm DNA integrity, expressed as DNA Fragmentation Index (DFI), in a large caseload of testicular cancer patients by sperm chromatin structure assay. The mean total DFI for all patients at T0 was 18.0 AE 12.5%. Sperm chromatin profile was markedly impaired at T3 (27.7 AE 17.4%) and T6 (23.2 AE 15.3%), improving considerably at T12 and T24 (14.0 AE 8.9% and 14.4 AE 10.3%). After chemotherapy, we found a marked increase in DFI at T3 and T6 and a significant reduction at T12 and T24 in comparison with the baseline. In contrast, DFI increased at T3 and T6 after radiotherapy but the subsequent reduction was far less marked, reaching baseline values at T12 and T24. Finally, post-treatment DNA damage was not age or histotype dependent, but was more marked in the advanced stage of cancer. In this study, we showed that the chromatin profile may be affected in the months immediately following the end of the treatment, improving after 12-24 months. Our results thus indicate that post-treatment DNA damage is influenced both by the type and intensity of the therapy and by the pathological and clinical stage of the disease.

Impact of chemotherapy and radiotherapy for testicular germ cell tumors on spermatogenesis and sperm DNA: a multicenter prospective study from the CECOS network

Fertility and Sterility, 2013

Objective: To determine the consequences of adjuvant testicular germ cell tumor treatment (TGCT) on sperm characteristics and sperm DNA, and to evaluate the predictors of sperm recovery. Design: Multicenter prospective longitudinal study of patients analyzed before treatment and after 3, 6, 12, and 24 months. Setting: University hospitals. Patient(s): One hundred twenty-nine volunteer TGCT patients and a control group of 257 fertile men. Intervention(s): Routine semen analyses, sperm DNA, and chromatin assessments. Main Outcome Measure(s): Comparisons of mean sperm characteristics before and after treatment, with sperm recovery analyzed by the Kaplan-Meier method. Result(s): The quantitative and qualitative sperm characteristics decreased after treatment, with lowest values at 3 and 6 months and with variations according to treatment type. The mean total sperm count recovered to pretreatment values at 12 months after treatment after two or fewer bleomycin, etoposide, and cisplatin (BEP) cycles, but not after radiotherapy or more than two BEP cycles. Only the treatment modalities and pretreatment sperm production were related to recovery of the World Health Organization reference sperm values. An increased proportion of patients had elevated high sperm DNA stainability at 6 months after radiotherapy.

The impact of testicular carcinoma and its treatment on sperm DNA integrity

Cancer, 2004

BACKGROUND. In patients with testicular germ cell carcinoma (TGCC), spermatogenesis and fertility are impaired. Intracytoplasmic sperm injection has improved their possibility of fatherhood, but might also impose a risk of transmitting DNA defects to the offspring. The aim of the current study was to evaluate the impact of chemotherapy and irradiation on sperm DNA integrity.