The Role Of Subclinical Infection And Cytokines In Prete

亚临床感染和细胞因子在 Prete 中的作用

• 批准号: 7334071
• 负责人: ROBERTO ROMERO
• 金额: --
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7334071
• 关键词: Role; Subclinical; Infection; Cytokines; Prete

Premature birth is the leading cause of perinatal mortality and morbidity worldwide. The Perinatology Research Branch has defined preterm labor as a syndrome and determined that at least 25% of preterm neonates are born to women with sub-clinical intrauterine infection. The Branch has also provided evidence that many premature neonates are critically ill before birth and proposed that, in the context of intrauterine infection, the onset of premature labor has survival value. The goal of this project is to understand the pathophysiology of premature labor and delivery. This year, our research focused on the following studies: 1. Gene expression signature of spontaneous term labor without histologic chorioamnionitis. Human parturition involves a common pathway manifested clinically by uterine contractions, cervical ripening and chorioamniotic membrane/decidual activation, culminating in membrane rupture. Previous studies have linked parturition to an inflammatory process. Investigators at the Branch analyzed both the transcriptome in the chorioamniotic membranes and maternal blood in order to identify the biological processes in normal spontaneous labor using an unbiased genome-wide approach. Transcriptional profiles for chorioamniotic membranes and blood were generated from patients at term with no labor and patients at term in labor. All placentas were subjected to histologic examination, and a criterion for inclusion was the absence of histologic chorioamnionitis. An increased expression of multiple chemokines and transcripts associated with neutrophil and monocyte recruitment was observed among patients in term labor. The results of this study indicate that labor induces gene expression changes consistent with localized inflammation despite the absence of histological chorioamnionitis. 2. Uterine transcriptomes of bacteria-induced and ovariectomy-induced preterm labor (PTL) in mice are characterized by differential expression of arachidonate metabolism genes. PTL is a syndrome with multiple etiologies. In this study, the Branch focused on two well established mechanisms of PTL: intrauterine infection and the requirement of progesterone for pregnancy maintenance. To investigate the role of intrauterine infection and progesterone in human PTL, investigators analyzed the uterine transcriptome of mice using two models: the first was the induction of PTL by intrauterine infection with Escherichia coli, and the second was the induction of PTL by ovariectomy and progesterone withdrawal. Quantitative reverse transcriptase?polymerase chain reaction measurements demonstrated that bacteria-induced PTL substantially increased the expression of genes involved in prostaglandin synthesis. In contrast, ovariectomy-induced PTL increased the expression of genes involved in lipoxin, leukotriene, and hydroxyeicosatetraenoic acid synthesis. The results of this study indicate that bacteria-induced and ovariectomy-induced PTL each express a different balance of genes that are involved in prostaglandin synthesis and the synthesis of lipoxins, leukotrienes, and HETEs. This balance may be important to PTL and is amenable to experimental investigation. 3. The transcriptome of the uterine cervix before and after spontaneous term parturition. Cervical ripening is one of the components of the common pathway for parturition. Therefore, determining the expression profile of genes involved in this process is crucial for the understanding of mechanisms leading to premature cervical ripening and preterm delivery. This study provided an unbiased and comprehensive description of the changes in the cervical transcriptome before and after spontaneous term labor. The transcriptome of cervical tissue from patients at term not in labor and after spontaneous labor were characterized using Affymetrix microarrays. Results of this study revealed that the cervical transcriptome of term patients who underwent labor is dramatically different from that of patients without labor, with 1192 differentially expressed genes. Among the up-regulated genes were those involved in neutrophil chemotaxis, apoptosis, extracellular matrix regulation, and steroid metabolism. Genes involved in neutrophil chemotaxis were up-regulated in specimens from women after spontaneous labor, and an increased expression of IL-8, IL-6, and scular endothelial growth factor was confirmed by real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Toll-like receptor (TLR)-3 and TLR-5 showed decreased gene expression in patients after spontaneous labor in the microarray analysis and this was confirmed by qRT-PCR. 4. A rapid bedside test for the prediction of preterm delivery (PTD). Intra-amniotic infection (IAI) and inflammation have been causally linked to preterm labor (PTL), PTD and fetal injury. Intra-amniotic inflammation can be detected by performing an amniocentesis and measuring cytokines (e.g. IL-6 and MMP-8), white blood cell count, and glucose concentrations in the amniotic fluid. It has been previously demonstrated that elevated MMP-8 concentrations in the amniotic fluid of patients with PTL and intact membranes can diagnose intra-amniotic inflammation with 95% sensitivity and 93% specificity. This year, the Branch conducted a study to investigate the diagnostic indices, predictive values, efficiency and likelihood ratios of the MMP-8 PTD Check test for the detection of IAI, inflammation, spontaneous PTD and severe neonatal morbidity among patients with increased uterine contractions and intact membranes. The MMP-8 PTD Check Test (SK Pharma Co, Ltd, Kyunggi-do, Korea) is a rapid bedside test (configured similarly to a rapid pregnancy test) designed to provide a quick bedside assessment for elevated concentrations of MMP-in a small amount of amniotic fluid. A positive MMP-8 rapid test correctly predicted 70% of the spontaneous PTDs within 48 hrs, and 94% of those occurring within 7 days and 14 days of the amniocentesis. The efficiency of a positive MMP-8 rapid test in the identification of IAI and inflammation was 94% (311/331) and 93% (308/331), respectively. We concluded that this rapid test gives clinicians a fast and accurate assessment of the inflammatory status of the amniotic cavity, and allows for better identification of patients at risk for impending PTD. Follow-up clinical trials are required to determine the role of the MMP-8 rapid test for the identification of inflammation at the time of amniocentesis, as well as determine whether, based on the rapid test results, treatment with antibiotics and/or anti-inflammatory agents may improve pregnancy outcome. 5. Phylogenetic analysis reveals the evolution of the mammalian placenta. The placenta is the lifeline for mammalian reproduction and a complex organ that provides clues about natural selection and evolution. Through phylogenetic analysis of molecular and anatomical data, investigators at the Branch presented evidence describing the evolutionary history of the placenta of eutherian mammals?the group that includes all mammal species except marsupials and the egg laying monotremes (for example, the duck billed platypus). In contrast to most theories, the study found that the disc-shaped, hemochorial placenta of many primates, including humans, existed throughout the eutherian lineage from the last common ancestor of placental mammals to the emergence of humans. That intimate contact between fetal and maternal blood was established in the last common ancestor of the crown group of Eutheria gives credence to the hypothesis that successful pregnancy requires appropriate allorecognition and tolerance at the maternal-fetal interface.

早产是全世界围产期死亡率和发病率的主要原因。围产学研究部门将早产定义为一种综合征，并确定至少 25% 的早产新生儿是患有亚临床宫内感染的妇女所生。该分会还提供了许多早产儿出生前病情危重的证据，并提出，在宫内感染的情况下，早产的发生具有生存价值。该项目的目标是了解早产和分娩的病理生理学。今年，我们的研究重点如下： 1. 无组织学绒毛膜羊膜炎的自发足月分娩的基因表达特征。人类分娩涉及一个常见的途径，临床上表现为子宫收缩、宫颈成熟和绒毛膜/蜕膜激活，最终导致胎膜破裂。先前的研究已将分娩与炎症过程联系起来。该分支机构的研究人员分析了绒毛膜羊膜和母血中的转录组，以便使用无偏见的全基因组方法来识别正常自发分娩的生物过程。绒毛膜羊膜和血液的转录谱来自未临产的足月患者和临产的足月患者。所有胎盘均接受组织学检查，纳入标准是不存在组织学绒毛膜羊膜炎。在足月分娩的患者中观察到与中性粒细胞和单核细胞募集相关的多种趋化因子和转录物的表达增加。这项研究的结果表明，尽管不存在组织学绒毛膜羊膜炎，但分娩会诱导与局部炎症一致的基因表达变化。 2.细菌诱导和卵巢切除诱导的小鼠早产（PTL）的子宫转录组的特征是花生四烯酸代谢基因的差异表达。 PTL 是一种具有多种病因的综合征。在这项研究中，该部门重点关注 PTL 的两个成熟机制：宫内感染和维持妊娠所需的黄体酮。为了研究宫内感染和黄体酮在人类 PTL 中的作用，研究人员使用两种模型分析了小鼠的子宫转录组：第一个是宫内感染大肠杆菌诱导 PTL，第二个是卵巢切除和黄体酮诱导 PTL撤回。定量逆转录酶聚合酶链式反应测量表明，细菌诱导的 PTL 显着增加了前列腺素合成相关基因的表达。相反，卵巢切除术诱导的 PTL 增加了参与脂氧素、白三烯和羟基二十碳四烯酸合成的基因的表达。这项研究的结果表明，细菌诱导的和卵巢切除诱导的 PTL 各自表达不同的基因平衡，这些基因参与前列腺素合成以及脂氧素、白三烯和 HETE 的合成。这种平衡对于 PTL 可能很重要，并且适合实验研究。 3.自然足月分娩前后子宫颈的转录组。宫颈成熟是分娩共同途径的组成部分之一。因此，确定参与这一过程的基因的表达谱对于理解导致宫颈早熟和早产的机制至关重要。这项研究对自然足月分娩前后宫颈转录组的变化进行了公正和全面的描述。使用 Affymetrix 微阵列对足月非临产患者和自然临产后患者的宫颈组织转录组进行表征。这项研究结果显示，足月临产患者的宫颈转录组与未临产患者的宫颈转录组存在显着差异，有 1192 个差异表达基因。上调的基因涉及中性粒细胞趋化性、细胞凋亡、细胞外基质调节和类固醇代谢。自然分娩后女性样本中涉及中性粒细胞趋化性的基因上调，实时定量逆转录酶聚合酶链反应 (qRT) 证实 IL-8、IL-6 和血管内皮生长因子表达增加。 -PCR）。在微阵列分析中，Toll 样受体 (TLR)-3​​ 和 TLR-5 显示自然分娩后患者的基因表达下降，并通过 qRT-PCR 证实了这一点。 4. 预测早产 (PTD) 的快速床边测试。羊膜内感染 (IAI) 和炎症与早产 (PTL)、PTD 和胎儿损伤有因果关系。羊膜内炎症可以通过进行羊膜穿刺术并测量羊水中的细胞因子（例如 IL-6 和 MMP-8）、白细胞计数和葡萄糖浓度来检测。先前已证明，PTL 和胎膜完整的患者羊水中 MMP-8 浓度升高可以诊断羊膜内炎症，敏感性为 95%，特异性为 93%。今年，该科开展了一项研究，探讨MMP-8 PTD Check测试的诊断指标、预测值、效率和似然比，以检测子宫收缩增强患者和新生儿中IAI、炎症、自发性PTD和严重新生儿发病率。完整的膜。 MMP-8 PTD 检查测试（韩国京畿道 SK Pharma Co., Ltd）是一种快速床边测试（配置与快速妊娠测试类似），旨在为小样本中 MMP 浓度升高提供快速床边评估。羊水量。 MMP-8 快速检测呈阳性，可正确预测 70% 的 48 小时内自发性 PTD，以及 94% 的羊膜穿刺术后 7 天和 14 天内发生的自发性 PTD。 MMP-8 阳性快速检测鉴定 IAI 和炎症的效率分别为 94% (311/331) 和 93% (308/331)。我们的结论是，这种快速检测可以让临床医生快速准确地评估羊膜腔的炎症状态，并可以更好地识别有即将发生 PTD 风险的患者。需要进行后续临床试验来确定 MMP-8 快速检测在羊膜穿刺术时识别炎症中的作用，并根据快速检测结果确定是否使用抗生素和/或抗炎药进行治疗。炎症剂可能会改善妊娠结局。 5. 系统发育分析揭示了哺乳动物胎盘的进化。胎盘是哺乳动物繁殖的生命线，也是一个复杂的器官，为自然选择和进化提供线索。通过分子和解剖数据的系统发育分析，该分部的研究人员提供了描述真兽类哺乳动物胎盘进化史的证据，真兽类哺乳动物包括除有袋动物和产蛋单孔类动物（例如鸭嘴兽）之外的所有哺乳动物物种。与大多数理论相反，该研究发现，包括人类在内的许多灵长类动物的盘状血绒胎盘存在于从胎盘哺乳动物的最后共同祖先到人类出现的整个真兽谱系中。胎儿和母体血液之间的密切接触是在真兽亚冠群的最后一个共同祖先中建立起来的，这证实了这样的假设：成功怀孕需要在母胎界面处进行适当的同种异体识别和耐受。