The Ronald O. Perelman and Claudia Cohen Center for Reproductive Medicine, Weill Cornell Medicine, 1305 York Avenue, Y720, New York, NY, 10021, USA.
J Assist Reprod Genet. 2021 Aug;38(8):2073-2086. doi: 10.1007/s10815-021-02191-4. Epub 2021 Apr 20.
To characterize, by specific biomarkers and nucleic acid sequencing, the structural and genomic sperm characteristics of partial (PG) and complete globozoospermic (CG) men in order to identify the best reproductive treatment.
We assessed spermatozoa from 14 consenting men ultrastructurally, as well as for histone content, sperm chromatin integrity, and sperm aneuploidy. Additional genomic, transcriptomic, and proteomic evaluations were carried out to further characterize the CG cohort. The presence of oocyte-activating sperm cytosolic factor (OASCF) was measured by a phospholipase C zeta (PLCζ) immunofluorescence assay. Couples were treated in subsequent cycles either by conventional ICSI or by ICSI with assisted gamete treatment (AGT) using calcium ionophore (Ionomycin, 19657, Sigma-Aldrich, Saint Louis, MO, USA).
Ultrastructural assessment confirmed complete acrosome deficiency in all spermatozoa from CG men. Histone content, sperm chromatin integrity, and sperm aneuploidy did not differ significantly between the PG (n = 4) and CG (n = 10) cohorts. PLCζ assessment indicated a positive presence of OASCF in 4 PG couples, who underwent subsequent ICSI cycles that yielded a 36.1% (43/119) fertilization with a 50% (2/4) clinical pregnancy and delivery rate. PLCζ assessment failed to detect OASCF for 8 CG patients who underwent 9 subsequent ICSI cycles with AGT, yielding a remarkable improvement of fertilization (39/97; 40.2%) (P = 0.00001). Embryo implantation (6/21; 28.6%) and clinical pregnancies (5/7; 71.4%) were also enhanced, resulting in 4 deliveries. Gene mutations (DPY19L2, SPATA16, PICK1) were identified in spermatozoa from CG patients. Additionally, CG patients unable to sustain a term pregnancy had gene mutations involved in zygote development (NLRP5) and postnatal development (BSX). CG patients who successfully sustained a pregnancy had a mutation (PIWIL1) related to sperm phenotype. PLCZ1 was both mutated and underexpressed in these CG patients, regardless of reproductive outcome.
Sperm bioassays and genomic studies can be used to characterize this gamete's capacity to support embryonic development and to tailor treatments maximizing reproductive outcome.
通过特定的生物标志物和核酸测序，对部分（PG）和完全球形精子（CG）男性的结构和基因组精子特征进行描述，以确定最佳的生殖治疗方法。
我们对 14 名同意的男性的精子进行了超微结构评估，以及组蛋白含量、精子染色质完整性和精子非整倍体检测。对 CG 队列进行了额外的基因组、转录组和蛋白质组评估，以进一步描述 CG 队列。通过 PLCζ 免疫荧光测定测量卵母细胞激活精子胞质因子 (OASCF) 的存在。用钙离子载体（离子霉素，19657，Sigma-Aldrich，圣路易斯，密苏里州，美国）处理夫妇，随后在常规 ICSI 或 ICSI 辅助配子处理（AGT）中进行后续周期的治疗。
超微结构评估证实所有 CG 男性的精子完全缺乏顶体。PG（n=4）和 CG（n=10）队列之间的组蛋白含量、精子染色质完整性和精子非整倍体没有显著差异。PLCζ 评估表明，在 4 对 PG 夫妇中存在阳性的 OASCF，他们随后进行了 ICSI 周期，受精率为 36.1%（43/119），临床妊娠率和分娩率为 50%（2/4）。PLCζ 评估未能检测到 8 名 CG 患者的 OASCF，他们随后进行了 9 次 ICSI 周期的 AGT，受精率显著提高（39/97；40.2%）（P=0.00001）。胚胎着床（6/21；28.6%）和临床妊娠（5/7；71.4%）也得到了增强，导致 4 例分娩。在 CG 患者的精子中发现了基因突变（DPY19L2、SPATA16、PICK1）。此外，无法维持足月妊娠的 CG 患者的基因突变为合子发育（NLRP5）和产后发育（BSX）所涉及。成功维持妊娠的 CG 患者的精子表型相关基因突变（PIWIL1）。无论生殖结果如何，这些 CG 患者的 PLCZ1 均发生突变和低表达。
精子生物测定和基因组研究可用于描述配子支持胚胎发育的能力，并针对最大限度提高生殖结果的治疗方法进行调整。
