氧张力调节滋养外胚层细胞释放的外泌体的 miRNA 谱和生物活性-用于监测妊娠并发症的液体活检。
Oxygen tension regulates the miRNA profile and bioactivity of exosomes released from extravillous trophoblast cells - Liquid biopsies for monitoring complications of pregnancy.
机构信息
Exosome Biology Laboratory, Centre for Clinical Diagnostics, University of Queensland Centre for Clinical Research, Royal Brisbane and Women's Hospital, The University of Queensland, Brisbane, Queensland, Australia.
Maternal-Fetal Medicine, Department of Obstetrics and Gynecology, Ochsner Clinic Foundation, New Orleans, United States of America.
出版信息
PLoS One. 2017 Mar 28;12(3):e0174514. doi: 10.1371/journal.pone.0174514. eCollection 2017.
Our understanding of how cells communicate has undergone a paradigm shift since the recent recognition of the role of exosomes in intercellular signaling. In this study, we investigated whether oxygen tension alters the exosome release and miRNA profile from extravillous trophoblast (EVT) cells, modifying their bioactivity on endothelial cells (EC). Furthermore, we have established the exosomal miRNA profile at early gestation in women who develop pre-eclampsia (PE) and spontaneous preterm birth (SPTB). HTR-8/SVneo cells were used as an EVT model. The effect of oxygen tension (i.e. 8% and 1% oxygen) on exosome release was quantified using nanocrystals (Qdot®) coupled to CD63 by fluorescence NTA. A real-time, live-cell imaging system (Incucyte™) was used to establish the effect of exosomes on EC. Plasma samples were obtained at early gestation (<18 weeks) and classified according to pregnancy outcomes. An Illumina TrueSeq Small RNA kit was used to construct a small RNA library from exosomal RNA obtained from EVT and plasma samples. The number of exosomes was significantly higher in EVT cultured under 1% compared to 8% oxygen. In total, 741 miRNA were identified in exosomes from EVT. Bioinformatic analysis revealed that these miRNA were associated with cell migration and cytokine production. Interestingly, exosomes isolated from EVT cultured at 8% oxygen increased EC migration, whilst exosomes cultured at 1% oxygen decreased EC migration. These changes were inversely proportional to TNF-α released from EC. Finally, we have identified a set of unique miRNAs in exosomes from EVT cultured at 1% oxygen and exosomes isolated from the circulation of mothers at early gestation, who later developed PE and SPTB. We suggest that aberrant exosomal signalling by placental cells is a common aetiological factor in pregnancy complications characterised by incomplete SpA remodeling and is therefore a clinically relevant biomarker of pregnancy complications.
自从最近发现外泌体在细胞间信号传递中的作用以来，我们对细胞间通讯的理解发生了重大转变。在这项研究中，我们研究了氧张力是否会改变滋养外胚层（EVT）细胞中外泌体的释放和 miRNA 谱，从而改变它们对内皮细胞（EC）的生物活性。此外，我们已经建立了在患有子痫前期（PE）和自发性早产（SPTB）的女性中早期妊娠的外泌体 miRNA 谱。HTR-8/SVneo 细胞被用作 EVT 模型。通过荧光 NTA 用与 CD63 偶联的纳米晶体（Qdot®）定量测量氧张力（即 8%和 1%氧气）对外泌体释放的影响。使用实时活细胞成像系统（Incucyte™）建立外泌体对 EC 的影响。在早期妊娠（<18 周）时获得血浆样本，并根据妊娠结局进行分类。使用 Illumina TrueSeq Small RNA 试剂盒从 EVT 和血浆样本中外泌体 RNA 构建小 RNA 文库。在 1%氧气下培养的 EVT 中外泌体的数量明显高于在 8%氧气下培养的 EVT。总共在 EVT 外泌体中鉴定出 741 个 miRNA。生物信息学分析表明，这些 miRNA 与细胞迁移和细胞因子产生有关。有趣的是，在 8%氧气下培养的 EVT 分离的外泌体增加了 EC 的迁移，而在 1%氧气下培养的 EVT 分离的外泌体减少了 EC 的迁移。这些变化与从 EC 释放的 TNF-α成反比。最后，我们在在 1%氧气下培养的 EVT 中外泌体和在早期妊娠母亲的循环中分离的外泌体中鉴定出一组独特的 miRNA，这些母亲后来发展为 PE 和 SPTB。我们认为，胎盘细胞异常的外泌体信号是不完全 SpA 重塑为特征的妊娠并发症的共同病因因素，因此是妊娠并发症的临床相关生物标志物。
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