Abstract

Background:

Abnormal placental development may result in adverse pregnancy outcomes and metabolic diseases in adulthood; however, it remains unknown whether and how xenobiotics affect human placentation.

Objectives:

This study aimed to screen and identify placentation-disrupting chemicals in commonly used organophosphate flame retardants (OPFRs) and, if identified, to investigate potential adverse effects on placentation in relation to adverse pregnancy outcomes and metabolic disorder in offspring in mice.

Methods:

We devised a high-throughput immunofluorescence screening assay based on human trophoblast organoids and used it to screen OPFRs that inhibit the proliferation of organoids. One identified chemical was assessed for its effects on placentation by evaluating villous cytotrophoblasts, syncytiotrophoblasts, and extravillous trophoblasts using immunofluorescence and a mitochondrial stress test after 2 d of exposure. A 10-d exposure study was further performed to observe the dynamic effect of the OPFR on the structure of the organoids. RNA-sequencing and western blotting experiments were performed to explore the associated pathways, and a potential binding protein was identified by immunoprecipitation and in vitro kinase activity assays. Animal studies were performed to determine whether the findings in organoids could be replicated in mice and to observe adverse pregnancy outcomes.

Results:

The proliferation of organoids exposed to three aryl-OPFRs was significantly lower than the proliferation of control organoids. Further analysis demonstrated that one such chemical, 2-ethylhexyl-diphenyl phosphate (EHDPP), disrupted placentation in organoids. Mechanistically, EHDPP interfered with insulin-like growth factor 1 receptor (IGF1R) to inhibit aerobic respiration. Mice exposed to EHDPP at a physiological human concentrations exhibited immature and mature placental disorders, which correlated with fetal growth restriction, implantation failure, stillbirth, and impaired glucose tolerance.

Conclusions:

The human trophoblast organoid model showed that the commonly used OPFRs disrupted placentation via IGF1R, indicating that its use may contribute to adverse pregnancy outcomes and metabolic disorders in offspring. https://doi.org/10.1289/EHP10273

中文翻译：

---

在人类滋养层类器官模型中筛选具有胎盘破坏作用的有机磷阻燃剂和小鼠不良妊娠结果的表征

摘要

背景：

胎盘发育异常可能导致成年期不良妊娠结局和代谢疾病；然而，目前尚不清楚外源性物质是否以及如何影响人类胎盘。

目标：

本研究旨在筛选和鉴定常用有机磷酸酯阻燃剂 (OPFR) 中的破坏胎盘的化学物质，如果确定，则调查与不良妊娠结局和小鼠后代代谢紊乱相关的对胎盘的潜在不利影响。

方法：

我们设计了一种基于人类滋养层类器官的高通量免疫荧光筛选试验，并用它来筛选抑制类器官增殖的 OPFR。通过在暴露 2 天后使用免疫荧光和线粒体压力测试评估绒毛细胞滋养细胞、合胞滋养细胞和绒毛外滋养细胞，评估了一种确定的化学物质对胎盘形成的影响。进一步进行了 10 天的暴露研究，以观察 OPFR 对类器官结构的动态影响。进行 RNA 测序和蛋白质印迹实验以探索相关途径，并通过免疫沉淀和体外鉴定潜在的结合蛋白激酶活性测定。进行动物研究以确定类器官中的发现是否可以在小鼠中复制并观察不良妊娠结果。

结果：

暴露于三种芳基-OPFRs 的类器官的增殖显着低于对照类器官的增殖。进一步的分析表明，其中一种化学物质 2-乙基己基二苯基磷酸酯 (EHDPP) 会破坏类器官中的胎盘形成。从机制上讲，EHDPP 干扰胰岛素样生长因子 1 受体 (IGF1R) 以抑制有氧呼吸。暴露于人体生理浓度 EHDPP 的小鼠表现出未成熟和成熟的胎盘疾病，这与胎儿生长受限、植入失败、死产和葡萄糖耐量受损有关。

结论：

人类滋养层类器官模型显示，常用的 OPFRs 通过 IGF1R 破坏胎盘，表明其使用可能导致不良妊娠结局和后代代谢紊乱。https://doi.org/10.1289/EHP10273