Parveen Arshiya, Mishra Suman, Srivastava Medha, Chaudhary Dharmendra K, Kapoor Deepa, Gupta Amrit, Tiwari Swasti
Department of Molecular Medicine & Biotechnology, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, India.
General Hospital, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, India.
Front Med (Lausanne). 2021 Dec 24;8:758971. doi: 10.3389/fmed.2021.758971. eCollection 2021.
Analysis of placental genes could unravel maternal-fetal complications. However, inaccessibility to placental tissue during early pregnancy has limited this effort. We tested if exosomes (Exo) released by human placenta in the maternal circulation harbor crucial placental genes. Placental alkaline phosphate positive exosomes (ExoPLAP) were enriched from maternal blood collected at the following gestational weeks; 6-8th (T1), 12-14th (T2), 20-24th (T3), and 28th-32nd (T4). Nanotracking analysis, electron microscopy, dynamic light scattering, and immunoblotting were used for characterization. We used microarray for transcriptome and quantitative PCR (qPCR) for gene analysis in ExoPLAP. Physical characterization and presence of CD63 and CD9 proteins confirmed the successful ExoPLAP enrichment. Four of the selected 36 placental genes did not amplify in ExoPLAP, while 32 showed regulations ( = 3-8/time point). Most genes in ExoPLAP showed significantly lower expression at T2-T4, relative to T1 ( < 0.05), such as , and . In contrast, genes, such as , and , had significantly higher expression at T2-T4 relative to T1. Unbiased gene profiling by microarray also confirmed expression of above genes in ExoPLAP-transcriptome. In addition, repeated measure ANOVA showed a significant change in the ExoPLAP transcriptome from T2 to T4 ( = 5/time point). Placental alkaline phosphate positive exosomes transcriptome changed with gestational age advancement in healthy women. The transcriptome expressed crucial placental genes involved in early embryonic development, such as actin cytoskeleton organization, appropriate cell positioning, DNA replication, and B-cell regulation for protecting mammalian fetuses from rejection. Thus, ExoPLAP in maternal blood could be a promising source to study the placental genes regulation for non-invasive monitoring of placental health.
胎盘基因分析有助于揭示母胎并发症。然而，孕早期难以获取胎盘组织限制了这一研究进展。我们检测了母体循环中人类胎盘释放的外泌体（Exo）是否携带关键胎盘基因。从以下孕周采集的母血中富集胎盘碱性磷酸酶阳性外泌体（ExoPLAP）：第6 - 8周（T1）、第12 - 14周（T2）、第20 - 24周（T3）以及第28 - 32周（T4）。采用纳米追踪分析、电子显微镜、动态光散射和免疫印迹进行表征。我们使用微阵列进行转录组分析，并通过定量PCR（qPCR）对ExoPLAP中的基因进行分析。CD63和CD9蛋白的物理表征及存在情况证实了ExoPLAP的成功富集。在所选的36个胎盘基因中，有4个在ExoPLAP中未扩增，而32个呈现出调控变化（每个时间点变化3 - 8倍）。与T1相比，ExoPLAP中的大多数基因在T2 - T4时表达显著降低（P < 0.05），如[具体基因1]、[具体基因2]和[具体基因3]。相反，[具体基因4]、[具体基因5]和[具体基因6]等基因在T2 - T4时相对于T1表达显著升高。微阵列的无偏基因谱分析也证实了上述基因在ExoPLAP转录组中的表达。此外，重复测量方差分析显示从T2到T4，ExoPLAP转录组有显著变化（每个时间点变化5倍）。健康女性中胎盘碱性磷酸酶阳性外泌体转录组随孕周进展而变化。该转录组表达了参与早期胚胎发育的关键胎盘基因，如肌动蛋白细胞骨架组织、适当的细胞定位、DNA复制以及B细胞调节，以保护哺乳动物胎儿免受排斥。因此，母血中的ExoPLAP可能是研究胎盘基因调控以无创监测胎盘健康的一个有前景的来源。
