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PGT-A后不同发育时间和评级囊胚的移植临床结局及转录组特征分析

Analysis of clinical outcomes and transcriptome characteristics of blastocysts with different developmental time and grades of PGT-A embryo transfer cycles
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摘要 目的探讨行胚胎植入前遗传学非整倍体检测(preimplantation genetic testing for aneuploidies,PGT-A)后不同发育时间和评级囊胚移植的临床结局,并对其转录组特征进行比较分析。方法回顾性队列研究分析2017年1月至2021年12月期间在山西省妇幼保健院生殖医学中心行PGT-A后选择整倍体囊胚移植的患者临床资料,共295个移植周期,分别按照囊胚发育时间[第5日(day 5,D5)组和第6日(day 6,D6)组]及囊胚评级(高质量组和一般质量组)进行分组,比较各组囊胚移植临床结局。通过比较来自GEO和ENA数据平台不同发育时间和评级的囊胚单细胞转录组数据(scRNA-seq),分析不同组间的转录组水平差异。结果①D5组和D6组男女方年龄、不孕类型、不孕年限、体质量指数(body mass index,BMI)、卵泡刺激素(follicle-stimulating hormone,FSH)、黄体生成素(luteinizing hormone,LH)、雌二醇和获卵数差异均无统计学意义(均P>0.05),两组间MⅡ卵率[86.35%(2051/2375)比82.71%(1770/2140),P=0.001]、囊胚形成率[68.08%(725/1065)比62.14%(540/869),P=0.006]、着床率[72.78%(115/158)比52.55%(72/137),P<0.001]、临床妊娠率[56.33%(89/158)比43.80%(60/137),P=0.032]和活产率[53.80%(85/158)比40.87%(56/137),P=0.027]相比较,D5组都显著高于D6组,两组间流产率、早产率、男性比例和出生体质量差异均没有统计学意义(均P>0.05);②高质量组和一般质量组男女方年龄、不孕类型、不孕年限、BMI、FSH、LH、雌二醇和获卵数差异均没有统计学意义(均P>0.05),两组间MⅡ卵率[87.06%(1251/1437)比83.50%(2570/3078),P=0.002]、囊胚形成率[73.38%(499/680)比61.08%(766/1254),P<0.001]、着床率[77.90%(74/95)比56.50%(113/200),P<0.001]、临床妊娠率[61.05%(58/95)比45.50%(91/200),P=0.013]和活产率[56.84%(54/95)比43.50%(87/200),P=0.032]相比较,高质量组都显著高于一般质量组,两组间流产率、早产率、新生儿男性比例和出生体质量差异均没有统计学意义(均P>0.05);③基于GEO和ENA数据平台的scRNA-seq数据,挖掘D5和D6囊胚以及高质量和一般质量囊胚内细胞团(inner cell mass,ICM)和外滋养层(trophectoderm,TE)的差异表达基因(differentially expressed genes,DEGs)。经过KEGG富集分析,显示D5组较D6组ICM/TE上调的DEGs显著富集在285/288个信号通路;高质量组较一般质量组ICM/TE上调的DEGs显著富集在207/3个信号通路。结论发育D5囊胚比发育D6囊胚,高质量囊胚比一般质量囊胚都具有较好的着床和继续临床妊娠能力。对不同发育时间和评级的囊胚进行转录组水平分析比较,显示转录组特征具有显著差异。囊胚转录组水平的分析,对囊胚着床及继续临床妊娠能力具有预测价值。 Objective To explore the clinical outcome of blastocysts with different developmental time and grades of preimplantation genetic testing for aneuploidies(PGT-A)embryo transfer cycles,and to compare and analyze their transcriptome characteristics.Methods The clinical data of patients with euploid blastocyst transplantation selected by PGT-A in Center for Reproductive Medicine of Women Health Center of Shanxi from January 2017 to December 2021 were retrospectively analyzed.A total of 295 transplantation cycles were divided into groups according to the day of embryo blastulation[day 5(D5)group and day 6(D6)group]and blastocyst grade(good-quality group and fair-quality group)and their clinical outcomes were compared.By comparing the single-cell RNA sequencing(scRNA-seq)data of blastocysts of different developmental time and grades from GEO and ENA data platforms,the transcriptome level differences among different groups were analyzed.Results 1)There were no significant differences in age of male and female,type of infertility,infertility duration,body mass index(BMI),follicle-stimulating hormone(FSH),luteinizing hormone(LH),estradiol and number of oocytes retrieved between D5 and D6 groups(all P>0.05).The MⅡoocyte rate[86.35%(2051/2375)vs.82.71%(1770/2140),P=0.001],blastocyst formation rate[68.08%(725/1065)vs.62.14%(540/869),P=0.006],implantation rate[72.78%(115/158)vs.52.55%(72/137),P<0.001],clinical pregnancy rate[56.33%(89/158)vs.43.80%(60/137),P=0.032]and live birth rate[53.80%(85/158)vs.40.87%(56/137),P=0.027]in D5 group were significantly higher than those in D6 group,and the results of miscarriage rate,preterm birth rate,proportion of male and birth weight between the two groups were not statistically significant.2)There were no significant differences in age of male and female,type of infertility,infertility duration,BMI,FSH,LH,estradiol and number of oocytes retrieved between good-quality and fair-quality groups(all P>0.05).The MⅡoocyte rate[87.06%(1251/1437)vs.83.50%(2570/3078),P=0.002],blastocyst formation rate[73.38%(499/680)vs.61.08%(766/1254),P<0.001],implantation rate[77.90%(74/95)vs.56.50%(113/200),P<0.001],clinical pregnancy rate[61.05%(58/95)vs.45.50%(91/200),P=0.013]and live birth rate[56.84%(54/95)vs.43.50%(87/200),P=0.032]in good-quality group were significantly higher than those in fair-quality group,and the results of miscarriage rate,preterm birth rate,proportion of male and birth weight between the two groups were not statistically significant.3)Based on the scRNA-seq data from GEO and ENA data platforms,we mined differentially expressed genes(DEGs)in the inner cell mass(ICM)and trophectoderm(TE)of D5 and D6 blastocysts,good-quality blastocysts and fair-quality blastocysts.Compared with D6 group,KEGG enrichment analysis showed that DEGs up-regulated of ICM/TE in D5 group were significantly enriched in 285/288 signaling pathways.DEGs up-regulated of ICM/TE were significantly enriched in 207/3 signaling pathways in the good-quality group compared with the fair-quality group.Conclusion In terms of implantation and clinical pregnancy ability,D5 blastocysts were better than D6 blastocysts,and good-quality blastocysts were better than fair-quality blastocysts.Transcriptome level analysis of blastocysts with different developmental time and grades showed significant differences in transcriptome characteristics.The analysis of blastocyst transcriptome level has predictive value for blastocyst implantation and clinical pregnancy ability.
作者 张磊 党诗莹 许素铭 张志平 范俊梅 张栋栋 武学清 Zhang Lei;Dang Shiying;Xu Suming;Zhang Zhiping;Fan Junmei;Zhang Dongdong;Wu Xueqing(Center of Reproductive Medicine,Children's Hospital of Shanxi,Women Health Center of Shanxi,Taiyuan 030000,China;Institutes of Biomedical Sciences,Shanxi University,Taiyuan 030000,China)
出处 《中华生殖与避孕杂志》 CAS CSCD 北大核心 2022年第11期1114-1120,共7页 Chinese Journal of Reproduction and Contraception
关键词 转录组 胚胎植入前遗传学非整倍体检测 囊胚发育时间 囊胚评级 临床结局 Transcriptome Preimplantation genetic testing for aneuploidies Day of embryo blastulation Blastocyst grade Clinical outcomes
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