1例PLCZ1复合杂合突变致IVF多原核受精及ICSI受精低下  

A case report on compound heterozygous mutations in PLCZ1 causing multiple pronuclei after IVF and poor fertilization after ICSI

作　　者：吴彤华 陈春媚 孙青 徐芬 莫美兰 WU Tonghua;CHEN Chunmei;SUN Qing;XU Fen;MO Meilan(Clinical Laboratory Center,Shenzhen Zhongshan Obstetrics&Gynecology Hospital,Formerly Shenzhen Zhongshan Urology Hospital,Shenzhen Zhongshan Institute for Reproductive Medicine and Genetics,Shenzhen Key Laboratory of Reproductive Immunology for Peri-implantation,Guangdong Engineering Technology Research Center of Reproductive Immunology for Peri-implantation,Shenzhen,Guangdong 518045,China;Reproductive Medicine Center,Shenzhen Zhongshan Obstetrics&Gynecology Hospital,Formerly Shenzhen Zhongshan Urology Hospital,Shenzhen,Guangdong 518045,China)

机构地区：[1]深圳中山妇产医院(原深圳中山泌尿外科医院)检验中心/深圳中山生殖与遗传研究所/深圳市围着床期生殖免疫重点实验室/广东省围着床期生殖免疫工程技术研究中心,广东深圳518045 [2]深圳中山妇产医院(原深圳中山泌尿外科医院)生殖医学中心,广东深圳518045

出　　处：《中国优生与遗传杂志》2024年第5期1034-1039,共6页Chinese Journal of Birth Health & Heredity

摘　　要：目的探讨一例继发不孕不育夫妻体外受精(IVF)中观察到大部分受精卵存在多原核(MPN)受精的遗传病因。方法该病例夫妻双方全血DNA样本行全外显子组测序。男方及其父母外周血DNA样本行磷脂酶Cζ1(PLCZ1)突变位点Sanger测序家系验证。成熟卵子在卵胞质内单精子注射(ICSI)后,行卵母细胞辅助激活(AOA)。结果女方未检出与MPN或受精障碍表型相关的变异。男方检出与受精障碍表型相关的1个致病性及1个疑似致病性变异,呈复合杂合,为PLCZ1(NM_033123.3)[c.588C>A/p.Cys196*];[c.590G>A/p.Arg197His]。患者配偶第二周期行拮抗剂方案促排卵,一半卵子行ICSI,正常受精率为12.50%(1/8),受精卵受精后凋亡;另一半卵子行ICSI+AOA,正常受精率为22.22%(2/9),形成4枚低等级卵裂胚。移植后未孕,囊胚培养失败。第三周期配偶行拮抗剂方案加生长激素促排卵,全部成熟卵子均行ICSI+AOA,正常受精率为44.44%(4/9),均形成优质胚胎。新鲜移植后单胎活产。这两个ICSI周期的受精卵均无MPN表型。结论PLCZ1基因缺陷的男性除受精失败和早期胚胎发育阻滞外,IVF时可呈现多精受精所致的多原核表型。采用ICSI+AOA的治疗策略可有效改善精子PLCZ1缺陷病例的受精情况及妊娠结局。Objective To investigate the genetic cause of multiple pronuclei(MPN) observed in most fertilized oocytes after in vitro fertilization(IVF) of a secondary infertile couple. Methods Whole-exome sequencing was performed on the peripheral blood DNA samples of this couple. Sanger sequencing confirmed the phospholipase C zeta 1(PLCZ1) variants on the peripheral blood DNA samples of the male and his parents. The mature oocytes underwent intracytoplasmic sperm injection(ICSI) followed by assisted oocyte activation(AOA). Results No mutations associated with MPN or fertilization failure phenotype was identified from the female patient of this case. One pathogenic and 1 likely pathogenic compound heterozygous variants related to fertilization failure phenotype were detected in the male partner, which were PLCZ1(NM_033123.3)[c.588C>A/p.Cys196~*];[c.590G>A/p.Arg197His]. Female partner was stimulated according to an antagonist protocol in the second cycle. Half oocytes were performed ICSI, and the 2PN fertilization rate was 12.50%(1/8). The only one fertilized oocyte was degenerated. The other half of the oocytes received ICSI-AOA treatment, and the 2PN fertilization rate was 22.22%(2/9) with four low-grade cleavage embryos formation. No pregnancy was achieved after embryo transfer, and no blastocyst development obtained after subsequently culture. In the third cycle, female partner underwent an antagonist protocol co-treatment within growth hormone, and ICSI+AOA was performed on all the mature oocytes obtained. The 2PN fertilization rate was 44.44%(4/9), and all embryos were good quality. A singleton live birth was achieved after fresh embryos transfer. No MPN phenotype was observed in all the fertilized zygotes from these two ICSI cycles. Conclusion In addition to fertilization failure and early embryo developmental arrest, men with PLCZ1 deficiency may present MPN genotype due to polyspermy during IVF. ICSI+AOA treatment strategy can effectively improve the fertilization and clinical outcome of spermic PLCZ1 defective case

关 键 词：PLCZ1 多原核 受精障碍 卵母细胞辅助激活 卵胞质内单精子注射 

分 类 号：R714.8[医药卫生—妇产科学]