摘要
目的分析体外受精-胚胎移植(IVF-ET)治疗后不同结局患者的血浆游离DNA(cfDNA)核小体印迹和差异基因表达谱,探讨影响妊娠结局及胚胎植入的差异基因的生物进程及细胞组分。方法收集2021年5月至2021年10月在广东医科大学顺德妇女儿童医院生殖中心的IVF-ET病例,根据胚胎植入后的结局分为失败组(n=5)和成功组(n=9),按胚胎植入前1天、植入后14天和植入后28天三个时间点分别检测患者血浆游离DNA,通过高通量测序对胚胎植入后不同结局和不同时间点的cfDNA核小体印迹和差异基因表达谱进行分析。结果血浆cfDNA核小体印迹中高表达基因的覆盖深度低于低表达基因,同一结局不同时间和同一时间不同结局的cfDNA核小体印迹的基因表达差异有统计学意义(P<0.05)。基于基因本体(GO)进行功能富集和信号通路分析发现,失败组在不同时间点的差异基因主要与免疫系统变化、发育调节和有机化合物的代谢相关,而成功组的与细胞发育和有机化合物的代谢相关。失败组差异表达基因在植入后14天的有机化合物代谢过程中表达下调,在植入后28天的免疫相关的生物过程表达上调。结论基于血浆游离DNA核小体印迹的差异基因表达分析可有效筛选和富集到胚胎植入后不同结局在不同时间点之间的差异表达基因和相关生物信号通路,为改善IVF-ET结局提供理论依据,利于研究和临床参考。
Objective To analyze the plasma cell-free DNA(cfDNA)nucleosome footprints and differential gene expression profiles of patients with different outcomes after In Vitro fertilization-embryo transfer(IVF-ET)treatment,and explore the biological processes and cellular components of differential genes that affect pregnancy outcomes and embryo implantation.Methods IVF-ET cases were collected in the Reproductive Center of Shunde Women and Children's Hospital of Guangdong Medical University from May 2021 to October 2021.They were divided into the failed group(n=5)and the pregnancy group(n=9)based on the outcome of embryo implantation.Plasma cfDNA was measured at three time:1 day before embryo implantation,14 days after embryo implantation,and 28 days after embryo implantation,analyzing cfDNA nucleosome footprints and differential gene expression profiles at different outcomes and different time points after embryo implantation by high-throughput sequencing.Results The coverage depth of high expression genes in plasma cfDNA nucleosome footprint was lower than that of low expression genes.The gene expression differences in cfDNA nucleosome footprint at different times of the same outcome and at different outcomes of the same time were statistically significant(P<0.05).Functional enrichment and signal pathway analysis based on Gene Ontology(GO)revealed that the differential genes in the failed group at different time points were mainly related to immune system changes,developmental regulation,and organic compound metabolism,while those in the pregnancy group were related to cell development and organic compound metabolism.The expression of genes in the failed group was down-regulated during the organic compound metabolism process in the 14 days after transfer,while the immune related biological processes expression was up-regulated at the 28 days after transfer.Conclusions Differential gene expression analysis based on plasma cell-free DNA nucleosome footprints can effectively screen and enrich differentially expressed genes
作者
梅凤琦
蔡晶
何珮清
刘彦慧
MEI Fengqi;CAI Jing;HE Peiqing;LIU Yanhui(Reproductive Medicine Center of Shunde Women and Children's Hospital of Guangdong Medical University,Foshan,Guangdong,China,528300;Medical Genetics Laboratory of Dongguan Maternal and Child Health Care Hospital,Dongguan,Guangdong,China,523057;Reproductive Medicine Center of Shenzhen Luohu People's Hospital,Shenzhen,Guangdong,China,51800)
出处
《分子诊断与治疗杂志》
2023年第12期2206-2210,共5页
Journal of Molecular Diagnostics and Therapy
