PDF(Pubmed)
Abstract：
OBJECTIVE: How do transcriptomics vary in haploid human androgenote embryos at single cell level in the first four cell cycles of embryo development?
CONCLUSIONS: Gene expression peaks at the fourth cell cycle, however some androcytes exhibit unique transcriptional behaviors.
BACKGROUND: The developmental potential of an embryo is determined by the competence of the oocyte and the sperm. However, studies of the contribution of the paternal genome using pure haploid androgenotes are very scarce.
METHODS: This study was performed analyzing the single-cell transcriptomic sequencing of 38 androcytes obtained from 10 androgenote bioconstructs previously produced in vitro (de Castro et al., 2023). These results were analyzed through different bioinformatics software such as g: Profiler, GSEA, Cytoscape, and Reactome.
METHODS: Single cell sequencing was used to obtain the transcriptomic profiles of the different androcytes. The results obtained were compared between the different cycles studied using the DESeq2 program and functional enrichment pathways using g: Profiler, Cytoscape, and Reactome.
RESULTS: A wave of paternally driven transcriptomic activation was found during the third-cell cycle, with 1128 upregulated and 225 downregulated genes and the fourth-cell cycle, with 1373 upregulated and 286 downregulated genes, compared to first-cell cycle androcytes. Differentially expressed routes related to cell differentiation, DNA-binding transcription, RNA biosynthesis and RNA polymerase II transcription regulatory complex, and cell death were found in the third and fourth with respect to the first-cell cycle. Conversely, in the fourth cell cycle, 153 downregulated and 332 upregulated genes were found compared with third cell cycle, associated with differentially expressed processes related to E-box binding and zinc finger protein 652 (ZNF652) transcription factor. Further, significant overexpression of LEUTX, PRAMEF1, DUXA, RFPL4A, TRIM43, and ZNF675 found in androgenotes, compared to biparental embryos, highlights the paternal contributions to zygote genome activation.
METHODS: All raw sequencing data are available through the Gene Expression Omnibus (GEO) under accessions number: GSE216501.
CONCLUSIONS: Extrapolation of biological events from uniparental constructs to biparental embryos should be done with caution. Maternal and paternal genomes do not act independently of each other in a natural condition. The absence of one genome may affect gene transcription of the other. In this sense, the haploid condition of the bioconstructs could mask the transcriptomic patterns of the single cells.
CONCLUSIONS: The results obtained demonstrated the level of involvement of the human paternal haploid genome in the early stages of embryo development as well as its evolution at the transcriptomic level, laying the groundwork for the use of these bioconstructs as reliable models to dispel doubts about the genetic role played by the paternal genome in the early cycles of embryo development.
BACKGROUND: This study was funded by Instituto de Salud Carlos III (ISCIII) through the project \'PI22/00924\', co-funded by European Regional Development Fund (ERDF); \'A way to make Europe\'. F.D. was supported by the Spanish Ministry of Economy and Competitiveness through the Miguel Servet program (CPII018/00002). M.J.E. was supported by Instituto de Salud Carlos III (PI19/00577 [M.J.E.]) and FI20/00086. P.dC. was supported by a predoctoral grant for training in research into health (PFIS PI19/00577) from the Instituto de Salud Carlos III. All authors declare having no conflict of interest with regard to this trial.
CONCLUSIONS: Gene expression peaks at the fourth cell cycle, however some androcytes exhibit unique transcriptional behaviors.
BACKGROUND: The developmental potential of an embryo is determined by the competence of the oocyte and the sperm. However, studies of the contribution of the paternal genome using pure haploid androgenotes are very scarce.
METHODS: This study was performed analyzing the single-cell transcriptomic sequencing of 38 androcytes obtained from 10 androgenote bioconstructs previously produced in vitro (de Castro et al., 2023). These results were analyzed through different bioinformatics software such as g: Profiler, GSEA, Cytoscape, and Reactome.
METHODS: Single cell sequencing was used to obtain the transcriptomic profiles of the different androcytes. The results obtained were compared between the different cycles studied using the DESeq2 program and functional enrichment pathways using g: Profiler, Cytoscape, and Reactome.
RESULTS: A wave of paternally driven transcriptomic activation was found during the third-cell cycle, with 1128 upregulated and 225 downregulated genes and the fourth-cell cycle, with 1373 upregulated and 286 downregulated genes, compared to first-cell cycle androcytes. Differentially expressed routes related to cell differentiation, DNA-binding transcription, RNA biosynthesis and RNA polymerase II transcription regulatory complex, and cell death were found in the third and fourth with respect to the first-cell cycle. Conversely, in the fourth cell cycle, 153 downregulated and 332 upregulated genes were found compared with third cell cycle, associated with differentially expressed processes related to E-box binding and zinc finger protein 652 (ZNF652) transcription factor. Further, significant overexpression of LEUTX, PRAMEF1, DUXA, RFPL4A, TRIM43, and ZNF675 found in androgenotes, compared to biparental embryos, highlights the paternal contributions to zygote genome activation.
METHODS: All raw sequencing data are available through the Gene Expression Omnibus (GEO) under accessions number: GSE216501.
CONCLUSIONS: Extrapolation of biological events from uniparental constructs to biparental embryos should be done with caution. Maternal and paternal genomes do not act independently of each other in a natural condition. The absence of one genome may affect gene transcription of the other. In this sense, the haploid condition of the bioconstructs could mask the transcriptomic patterns of the single cells.
CONCLUSIONS: The results obtained demonstrated the level of involvement of the human paternal haploid genome in the early stages of embryo development as well as its evolution at the transcriptomic level, laying the groundwork for the use of these bioconstructs as reliable models to dispel doubts about the genetic role played by the paternal genome in the early cycles of embryo development.
BACKGROUND: This study was funded by Instituto de Salud Carlos III (ISCIII) through the project \'PI22/00924\', co-funded by European Regional Development Fund (ERDF); \'A way to make Europe\'. F.D. was supported by the Spanish Ministry of Economy and Competitiveness through the Miguel Servet program (CPII018/00002). M.J.E. was supported by Instituto de Salud Carlos III (PI19/00577 [M.J.E.]) and FI20/00086. P.dC. was supported by a predoctoral grant for training in research into health (PFIS PI19/00577) from the Instituto de Salud Carlos III. All authors declare having no conflict of interest with regard to this trial.
摘要：
目的：在胚胎发育的前四个细胞周期中，单倍体人类雄激素胚胎在单细胞水平上的转录组学是如何变化的？
结论：基因表达在第四个细胞周期达到峰值，然而，一些雄激素表现出独特的转录行为。
背景：胚胎的发育潜力取决于卵母细胞和精子的能力。然而,使用纯单倍体雄激素对父本基因组的贡献的研究非常缺乏。
方法：本研究分析了从先前在体外产生的10个雄激素生物构建体获得的38个雄激素的单细胞转录组测序（deCastro等人。,2023年）。这些结果通过不同的生物信息学软件进行分析，如g：Profiler，GSEA,Cytoscape,和Reactome。
方法:使用单细胞测序来获得不同雄性腺细胞的转录组概况。在使用DESeq2程序研究的不同循环和使用g：Profiler的功能富集途径之间比较了获得的结果，Cytoscape,和Reactome。
结果：在第三个细胞周期中发现了父系驱动的转录组激活波，1128个上调和225个下调的基因和第四个细胞周期，有1373个上调的基因和286个下调的基因，与第一个细胞周期的雄激素相比。与细胞分化相关的差异表达途径，DNA结合转录，RNA生物合成和RNA聚合酶II转录调控复合物，在第一个细胞周期的第三个和第四个发现了细胞死亡。相反，在第四个细胞周期中，发现153个下调基因和332个上调基因与第三个细胞周期相比，与E盒结合和锌指蛋白652（ZNF652）转录因子相关的差异表达过程相关。Further,LEUTX的显著过表达，PRAMEF1,DUXA,RFPL4A,在雄激素中发现的TRIM43和ZNF675，与双亲胚胎相比，强调了父系对合子基因组激活的贡献。
方法:所有原始测序数据可通过基因表达综合(GEO)获得,材料号:GSE216501。
结论：从单亲构建体到双亲胚胎的生物学事件外推应该谨慎。母亲和父亲的基因组在自然条件下并不相互独立。一个基因组的缺失可能影响另一个基因组的基因转录。在这个意义上,生物构建体的单倍体条件可以掩盖单细胞的转录组模式。
结论：获得的结果表明，人类父系单倍体基因组参与胚胎发育的早期阶段以及转录组水平的进化，为使用这些生物构建体作为可靠的模型奠定基础，以消除对父系基因组在胚胎发育早期周期中所起的遗传作用的怀疑。
背景：这项研究由SaludCarlosIII研究所（ISCIII）通过项目“PI22/00924”资助，由欧洲区域发展基金(ERDF)共同资助；“一种制造欧洲的方式”。F.D.通过MiguelServet计划（CPII018/00002）得到西班牙经济和竞争力部的支持。M.J.E.得到了SaludCarlosIII研究所（PI19/00577[M.J.E.]）和FI20/00086的支持。P.dC.得到了SaludCarlosIII研究所的博士前资助，用于健康研究培训（PFISPI19/00577）。所有作者都声明与该试验没有利益冲突。
结论：基因表达在第四个细胞周期达到峰值，然而，一些雄激素表现出独特的转录行为。
背景：胚胎的发育潜力取决于卵母细胞和精子的能力。然而,使用纯单倍体雄激素对父本基因组的贡献的研究非常缺乏。
方法：本研究分析了从先前在体外产生的10个雄激素生物构建体获得的38个雄激素的单细胞转录组测序（deCastro等人。,2023年）。这些结果通过不同的生物信息学软件进行分析，如g：Profiler，GSEA,Cytoscape,和Reactome。
方法:使用单细胞测序来获得不同雄性腺细胞的转录组概况。在使用DESeq2程序研究的不同循环和使用g：Profiler的功能富集途径之间比较了获得的结果，Cytoscape,和Reactome。
结果：在第三个细胞周期中发现了父系驱动的转录组激活波，1128个上调和225个下调的基因和第四个细胞周期，有1373个上调的基因和286个下调的基因，与第一个细胞周期的雄激素相比。与细胞分化相关的差异表达途径，DNA结合转录，RNA生物合成和RNA聚合酶II转录调控复合物，在第一个细胞周期的第三个和第四个发现了细胞死亡。相反，在第四个细胞周期中，发现153个下调基因和332个上调基因与第三个细胞周期相比，与E盒结合和锌指蛋白652（ZNF652）转录因子相关的差异表达过程相关。Further,LEUTX的显著过表达，PRAMEF1,DUXA,RFPL4A,在雄激素中发现的TRIM43和ZNF675，与双亲胚胎相比，强调了父系对合子基因组激活的贡献。
方法:所有原始测序数据可通过基因表达综合(GEO)获得,材料号:GSE216501。
结论：从单亲构建体到双亲胚胎的生物学事件外推应该谨慎。母亲和父亲的基因组在自然条件下并不相互独立。一个基因组的缺失可能影响另一个基因组的基因转录。在这个意义上,生物构建体的单倍体条件可以掩盖单细胞的转录组模式。
结论：获得的结果表明，人类父系单倍体基因组参与胚胎发育的早期阶段以及转录组水平的进化，为使用这些生物构建体作为可靠的模型奠定基础，以消除对父系基因组在胚胎发育早期周期中所起的遗传作用的怀疑。
背景：这项研究由SaludCarlosIII研究所（ISCIII）通过项目“PI22/00924”资助，由欧洲区域发展基金(ERDF)共同资助；“一种制造欧洲的方式”。F.D.通过MiguelServet计划（CPII018/00002）得到西班牙经济和竞争力部的支持。M.J.E.得到了SaludCarlosIII研究所（PI19/00577[M.J.E.]）和FI20/00086的支持。P.dC.得到了SaludCarlosIII研究所的博士前资助，用于健康研究培训（PFISPI19/00577）。所有作者都声明与该试验没有利益冲突。
