Laboratory of Stem Cell & Cancer Epigenetic Research, School of Dentistry, UCLA, Los Angeles, California, United States of America.
CSUN-UCLA Stem Cell Research Bridge Program, Department of Biology, California State University at Northridge, Northridge, California, United States of America.
PLoS One. 2020 Nov 16;15(11):e0242276. doi: 10.1371/journal.pone.0242276. eCollection 2020.
Alcohol (ethanol, EtOH) consumption during pregnancy can result in fetal alcohol spectrum disorders (FASDs), which are characterized by prenatal and postnatal growth restriction and craniofacial dysmorphology. Recently, cell-derived extracellular vesicles, including exosomes and microvesicles containing several species of RNAs (exRNAs), have emerged as a mechanism of cell-to-cell communication. However, EtOH's effects on the biogenesis and function of non-coding exRNAs during fetal development have not been explored. Therefore, we studied the effects of maternal EtOH exposure on the composition of exosomal RNAs in the amniotic fluid (AF) using rat fetal alcohol exposure (FAE) model. Through RNA-Seq analysis we identified and verified AF exosomal miRNAs with differential expression levels specifically associated with maternal EtOH exposure. Uptake of purified FAE AF exosomes by rBMSCs resulted in significant alteration of molecular markers associated with osteogenic differentiation of rBMSCs. We also determined putative functional roles for AF exosomal miRNAs (miR-199a-3p, miR-214-3p and let-7g) that are dysregulated by FAE in osteogenic differentiation of rBMSCs. Our results demonstrate that FAE alters AF exosomal miRNAs and that exosomal transfer of dysregulated miRNAs has significant molecular effects on stem cell regulation and differentiation. Our results further suggest the usefulness of assessing molecular alterations in AF exRNAs to study the mechanisms of FAE teratogenesis that should be further investigated by using an in vivo model.
怀孕期间饮酒(乙醇,EtOH)可导致胎儿酒精谱系障碍(FASD),其特征为产前和产后生长受限以及颅面畸形。最近,细胞衍生的细胞外囊泡,包括含有几种 RNA(exRNA)的外泌体和微泡,已成为细胞间通讯的一种机制。然而,EtOH 对胎儿发育中非编码 exRNA 的生物发生和功能的影响尚未得到探索。因此,我们使用大鼠胎儿酒精暴露(FAE)模型研究了母体 EtOH 暴露对羊水(AF)中细胞外囊泡 RNA 组成的影响。通过 RNA-Seq 分析,我们鉴定并验证了与母体 EtOH 暴露特异性相关的具有差异表达水平的 AF 细胞外体 miRNAs。纯化的 FAE AF 细胞外体被 rBMSCs 摄取,导致与 rBMSCs 成骨分化相关的分子标志物发生显著改变。我们还确定了由 FAE 失调的 AF 细胞外体 miRNAs(miR-199a-3p、miR-214-3p 和 let-7g)在 rBMSCs 成骨分化中的潜在功能作用。我们的结果表明,FAE 改变了 AF 细胞外体 miRNAs,并且失调的 miRNAs 的细胞外体转移对干细胞调节和分化具有显著的分子影响。我们的结果进一步表明,评估 AF exRNAs 中的分子改变用于研究 FAE 致畸的机制是有用的,应进一步通过使用体内模型进行研究。