Ackerman William E, Rigo Mauricio M, DaSilva-Arnold Sonia C, Do Catherine, Tariq Mariam, Salas Martha, Castano Angelica, Zamudio Stacy, Tycko Benjamin, Illsley Nicholas P
Department of Obstetrics and Gynecology and AI.Health4All Center for Health Equity Using Machine Learning and Artificial Intelligence, University of Illinois College of Medicine, Chicago, USA.
Hackensack Meridian Health Center for Discovery and Innovation, Nutley, NJ.
bioRxiv. 2024 Jul 4:2024.07.02.601748. doi: 10.1101/2024.07.02.601748.
The phenotype of human placental extravillous trophoblast (EVT) at the end of pregnancy reflects both first trimester differentiation from villous cytotrophoblast (CTB) and later gestational changes, including loss of proliferative and invasive capacity. Invasion abnormalities are central to two major placental pathologies, preeclampsia and placenta accreta spectrum, so characterization of the corresponding normal processes is crucial. In this report, our gene expression analysis, using purified human CTB and EVT cells, highlights an epithelial-mesenchymal transition (EMT) mechanism underlying CTB-EVT differentiation and provides a trophoblast-specific EMT signature. In parallel, DNA methylation profiling shows that CTB cells, already hypomethylated relative to non-trophoblast cell lineages, show further genome-wide hypomethylation in the transition to EVT. However, a small subgroup of genes undergoes gains of methylation (GOM) in their regulatory regions or gene bodies, associated with differential mRNA expression (DE). Prominent in this GOM-DE group are genes involved in the EMT, including multiple canonical EMT markers and the EMT-linked transcription factor , for which we demonstrate a functional role in modulating the migratory and invasive capacities of JEG3 trophoblast cells. This analysis of DE associated with locus-specific GOM, together with functional studies of an important GOM-DE gene, highlights epigenetically regulated genes and pathways acting in human EVT differentiation and invasion, with implications for obstetric disorders in which these processes are dysregulated.
妊娠末期人胎盘绒毛外滋养层细胞（EVT）的表型既反映了孕早期从绒毛细胞滋养层细胞（CTB）分化而来的情况，也反映了后期妊娠变化，包括增殖和侵袭能力的丧失。侵袭异常是两种主要胎盘病理状态（先兆子痫和胎盘植入谱系疾病）的核心，因此对相应正常过程的特征描述至关重要。在本报告中，我们使用纯化的人CTB和EVT细胞进行基因表达分析，突出了CTB-EVT分化背后的上皮-间质转化（EMT）机制，并提供了滋养层细胞特异性的EMT特征。同时，DNA甲基化谱分析表明，相对于非滋养层细胞谱系，CTB细胞已经处于低甲基化状态，在向EVT转变过程中表现出全基因组范围的进一步低甲基化。然而，一小部分基因在其调控区域或基因体内发生甲基化增加（GOM），与mRNA表达差异（DE）相关。在这个GOM-DE组中突出的是参与EMT的基因，包括多个经典的EMT标志物和与EMT相关的转录因子，我们证明其在调节JEG3滋养层细胞的迁移和侵袭能力方面具有功能作用。这种与位点特异性GOM相关的DE分析，以及对一个重要的GOM-DE基因的功能研究，突出了在人EVT分化和侵袭中起作用的表观遗传调控基因和途径，对这些过程失调的产科疾病具有启示意义。
