Placental extracellular vesicles-associated microRNA-519c mediates endotoxin adaptation in pregnancy.
Placental extracellular vesicles-associated microRNA-519c mediates endotoxin adaptation in pregnancy.
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DOI：
10.1016/j.ajog.2021.06.075
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发表时间：
2021-12
影响因子：
9.8
通讯作者：
Hanna N
中科院分区：
文献类型：
--
作者：
Tiozzo C;Bustoros M;Lin X;Manzano De Mejia C;Gurzenda E;Chavez M;Hanna I;Aguiari P;Perin L;Hanna N
妊娠对母胎免疫界面是一项独特的挑战，需要在免疫抑制（这对维持半同种异体胎儿至关重要）和促炎宿主防御（以保护母胎界面免受入侵生物的侵害）之间取得平衡。对反复炎症刺激的适应（内毒素耐受）对于防止因母体对孕期常见的轻度/中度感染产生过度炎症反应而导致的炎症诱导早产可能至关重要。然而，维持对反复感染耐受的确切机制尚未完全清楚。微小RNA（miRNA）在妊娠中起重要作用，有几种miRNA与妊娠组织功能以及病理妊娠状况有关。miRNA - 519c是C19MC簇的成员，是一种主要在胎盘中表达的人类特异性miRNA。然而，其在妊娠中的作用在很大程度上是未知的。
为了探究“内毒素耐受”失败在炎症介导的早产中常见的过度炎症反应发病机制中的作用。在本研究中，我们研究了胎盘特异性miRNA - 519c作为母胎界面内毒素耐受的关键调节因子的作用。
利用胎盘外植体培养系统，对足月和孕中期胎盘样本用脂多糖（LPS）进行处理。24小时后，收集条件培养基进行分析，并将胎盘外植体再次暴露于重复剂量的LPS中3天。分析上清液中的炎症标志物、细胞外囊泡（EVs）的存在以及微小RNA。为了研究微小RNA可能的作用机制，我们使用微小RNA模拟物和磷酸二酯酶3B（PDE3B）小干扰RNA（siRNA）转染评估了参与肿瘤坏死因子 - α（TNF - α）产生的PDE3B途径。最后，我们分析了来自不同孕周以及受炎症相关妊娠影响的女性的人胎盘样本。
我们的数据显示，人胎盘反复暴露于内毒素刺激会诱导一种耐受表型，其特征是TNF - α降低和白细胞介素 - 10（IL - 10）水平上调。这种反应是由胎盘细胞外囊泡内包裹的胎盘特异性miRNA - 519c介导的。LPS处理增加了对外泌体四跨膜蛋白标志物（即CD9、CD63和CD81）呈阳性的细胞外囊泡，这些囊泡主要由滋养层细胞分泌。用miR - 519c模拟物转染的原代人滋养层细胞中PDE3B降低。而通过siRNA转染使PDE3B缺失会导致TNF - α产生减少。这些数据支持了miRNA - 519c的抗炎作用是通过下调磷酸二酯酶3B途径，从而抑制TNF - α产生这一假设。此外，来自正常妊娠和炎症相关妊娠的人胎盘表明，胎盘miRNA - 519c水平降低与孕期感染诱导的炎症病变有关。
我们确定了miRNA - 519c（一种人胎盘特异性miRNA）是母胎界面上与感染诱导早产相关的免疫适应的新型调节因子。我们的研究可为未来探索miRNA - 519c作为感染诱导早产的生物标志物的潜在用途的实验提供基础。
胎盘特异性miRNA - 519c在人类妊娠的内毒素耐受中起关键作用，并提示其可能用作宫内炎症疾病的生物标志物。
Pregnancy represents a unique challenge for the maternal-fetal immune interface, requiring a balance between immunosuppression, which is essential for the maintenance of a semi-allogeneic fetus, and pro-inflammatory host defense to protect the maternal-fetal interface from invading organisms. Adaptation to repeated inflammatory stimuli (endotoxin tolerance) may be critical in preventing inflammation-induced preterm birth resulting from exaggerated maternal inflammatory responses to mild/moderate infections that are common during pregnancy. However, the exact mechanisms contributing to the maintenance of tolerance to repeated infections are not completely understood. miRNAs play important roles in pregnancy, with several miRNAs implicated in gestational tissue function, as well as in pathologic pregnancy conditions. miRNA-519c, a member of the C19MC cluster, is a human-specific miRNA mainly expressed in the placenta. However, its role in pregnancy is largely unknown. To explore the role of “endotoxin tolerance” failure in the pathogenesis of an exaggerated inflammatory response often seen in inflammation-mediated preterm birth. In this study, we investigated the role of miRNA-519c, a placenta-specific miRNA, as a key regulator of endotoxin tolerance at the maternal-fetal interface. Using a placental explants culture system, samples from term and 2nd-trimester placentas were treated with LPS. After 24 hours, the conditioned media was collected for analysis, and the placental explants were re-exposed to repeated doses of LPS for 3 days. The supernatant was analyzed for inflammatory markers, presence of extracellular vesicles (EVs) and microRNAs. To study the possible mechanism of action of the microRNAs, we evaluated the phosphodiesterase 3 B (PDE3B) pathway involved in TNF-α production using a miRNAs mimic and PDE3B siRNA transfection. Finally, we analyzed human placental samples from different gestational ages and from women affected by inflammation-associated pregnancies. Our data showed that repeated exposure of the human placenta to endotoxin challenges induced a tolerant phenotype characterized by decreased TNF-α and upregulated IL-10 levels. This reaction was mediated by the placenta-specific miRNA-519c packaged within placental EVs. LPS treatment increased the EVs that were positive for the exosome tetraspanin markers, namely CD9, CD63, and CD81, and secreted primarily by trophoblasts. Primary human trophoblast cells transfected with miR-519c mimic decreased PDE3B. While lack of PDE3B, achieved by siRNA transfection, resulted in a decreased TNF-α production. These data supported the hypothesis that the anti-inflammatory action of miRNA-519c was mediated by a downregulation of the phosphodiesterase 3 B pathway, leading to inhibition of TNF-α production. Furthermore, human placentas from normal and inflammation-associated pregnancies demonstrated that decreased placental miRNA-519c level was linked to infection-induced inflammatory pathologies during pregnancy. We identified miRNA-519c, a human placenta-specific miRNA, as a novel regulator of immune adaptation associated with infection-induced preterm birth at the maternal-fetal interface. Our study can serve as a basis for future experiments to explore the potential use of miRNA-519c as a biomarker for infection-induced preterm birth. Placenta-specific miRNA-519c plays a critical role in endotoxin tolerance in human pregnancy and suggest its possible use as a biomarker for intrauterine inflammatory disorders.
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DOI：
10.1111/aji.12623
发表时间：
2017-03
期刊：
American journal of reproductive immunology (New York, N.Y. : 1989)
影响因子：
--
作者：
Anders AP;Gaddy JA;Doster RS;Aronoff DM
通讯作者：
Aronoff DM
影响因子：
3.2
作者：
Gisslen, Tate;Hillman, Noah H.;Kallapur, Suhas G.
通讯作者：
Kallapur, Suhas G.
DOI：
10.1111/j.1600-0897.2011.00995.x
发表时间：
2011-10-01
影响因子：
3.6
作者：
Atay, Safinur;Gercel-Taylor, Cicek;Taylor, Douglas D.
通讯作者：
Taylor, Douglas D.
影响因子：
14.9
作者：
Bortolin-Cavaillé ML;Dance M;Weber M;Cavaillé J
通讯作者：
Cavaillé J
影响因子：
3.8
作者：
Dumont, Tina M. F.;Mouillet, Jean-Francois;Sadovsky, Yoel
通讯作者：
Sadovsky, Yoel