Outcomes Of ICSI/PGD And Relative Analysis In Patients With Klinefelter Syndrome And Globozoospermia

Section 1Embryo and pregnancy outcomes of ICSI-PGD with ejaculated spermatozoa in 14 patients with Klinefelter syndromeBackground Klinefelter syndrome (KS) is characterized of azoospermia, and oligoasthenozoospermia in minority patients, the technique of intracytoplasmic sperm injection(ICSI) with testicular sperm extraction(TESE) or ejaculated sperm helps many KS patients father their own babies. With high proportion of abnormal spermatozoa and embryos, the application of preimplantation genetic diagnosis (PGD) with fluorescent in situ hybridization (FISH) technology is recommended by many scholars to increase pregnancy rate and reduce the risk of birth defects. But with more and more healthy babies born through ICSI without PGD, along with the fact that the majority of spermatozoa of KS patients are normal, some researchers began to query the necessity of the application of PGD in KS patients.Objective This study aims to analyze the embryo and pregnancy outcomes of ICSI-PGD with ejaculated spermatozoa in KS patients and investigate the necessity of PGD for them.Materials and Methods From 2012 to 2015,14 KS patients were treated with ICSI-PGD with ejaculated spermatozoa. The pregnancy and embryo results of 15 cycles were analyzed and compared with the outcomes of ICSI with or without PGD in KS patients in literature.Outcomes1. In 115 embryos with explicit detection outcomes, there were 57 with XX, more than those with XY (43), and 15 with abnormal sex chromosomal signals (13%).2. In 16 embryo transfer cycles (1.54±0.72 embryos per transfer),14 implanted embryos (50% per transferred embryo) and 11 clinical pregnancies (68.75% per transfer) were obtained with births of 3 healthy boys and 8 healthy girls,3 ongoing clinical pregnancies.3. The proportion of embryos with sex chromosomal abnormalities is comparable with those (13.3%,17.9% respectively) in KS patients detected by PGD in literature (P=0.959, P=0.723 respectively), but much higher than that (3.4%) in infertile population under 35. However, there is no statistically significant difference in implantation rate and clinical pregnancy rate between ICSI without and with PGD in KS patients.Conclusions The impact of higher rate of abnormal embryos on pregnancy outcomes is rather limited and the application of PGD still plays a role considering the risk of miscarriage. More cases and follow-ups are needed to evaluate the benefits and potential risks of all ART methods for KS patients.Section 2Fertilization and pregnancy outcomes of ICSI in 6 patients with Globozoospermia and its relationship with DPY19L2Background Globozoospermia is one kind of severe teratozoospermia which occurs in less than 1% of infertile men. The fertilization rate and pregnancy rate are extremely low with conventional intracytoplasmic sperm injection (ICSI) treatment in globozoospermic patients (GP) due to the deletion or decrease of PCLξ which is a sperm-specific important factor for ovarian activation. The application of assisted ovarian activation (AOA) can facilitate ovarian activation and fertilization. And sperm with acrosomal residue can be selected during ICSI in some patients with motile sperm organellar morphology examination (MSOME) or transmission electron microscopy (TEM) or other techniques with high resolution, then IMSI was performed for these patients and live births were achieved. However, no studies were reported on the association between different genotypes and the outcomes of different ART methods.Objective This study aims to investigate the effect of different ART methods (ICSI or ICSI+AOA or IMSI) on pregnancy outcomes and the association between different genotypes of DPY19L2 and outcomes of different ART methods.Materials and Methods DPY19L2 gene sequencing was conducted in 6 GPs recruited in our hospital and their clinical data were collected. And the reproductive outcomes were compared according to different ART methods or genotypes of DPY19L2, including fertilization rate, cleavage rate, rate of D3 high-quality embryos and clinical pregnancy rate.Outcomes1. Mutations of DPY19L2 occurred in 4 out of 6 patients (66.7%),3 with complete deletion of the gene and 1 with a point mutation (c.869G>A).2. For patients with total globozoospermia, regardless of the presence of mutation of DPY19L2, conventional ICSI can’t achieve fertilization at all. And the fertilization rate is 44.4% with ICSI+AOA, while the rate of high-quality embryos is 62.5%. For patients with acrosomal residue found during ICSI, IMSI was performed and the fertilization rate is 20% and the rate of high-quality embryos is 66.7%. Patients with partial globozoospermia had the fertilization rate 62.5% and embryonic development. But no clinical pregnancy was obtained for all these patients.3. For patients with total globozoospermia and the point mutation, fertilization rate with ICSI+AOA and IMSI is 87.5% and 33.3% respectively; while those with DPY19L2 completely deleted had a rather low fertilization rate (0%,11.1%).Conclusions The application of AOA can largely improve the fertilization rate. And outcomes of ART have an association with different genotypes and phenotypes. Regardless of the use of AOA, ART outcomes are evidently better in patients with DPY19L2 point mutations or partial globozoospermia than those with total types and DPY19L2 deleted.