Targeted Inhibition of Interleukin-1 beta to Prevent Preterm Birth

项目概要/摘要 早产仍然是新生儿发病率和死亡率的一个重要原因。羊膜内感染， 炎症是早期早产的重要触发因素，其与由炎症介导的胎儿损伤有关。 羊水和胎儿中的细胞因子和其他炎症介质。本提案的目的是 确定新型选择性变构白细胞介素-1受体（IL-1 R）抑制剂的疗效和安全性 （Rytvela）作为产前治疗策略，以预防胎儿损伤和PTB。在怀孕的小鼠中， 强有力的证据表明，Rytvela是一种有效的抑制早产和胎儿损伤诱导的 脂多糖或脂儿茶酸。为了将这些发现转化为人类怀孕，我们 拟在妊娠非人灵长类动物模型中检测Rytvela的疗效并测定其药代动力学 B族链球菌（GBS）是一种临床上重要的细菌， 并可导致早产和新生儿败血症。我们独特的长期插管的非人类灵长类动物 在所有动物模型中，该模型与人类妊娠的相关性最大， 无法进入的隔室（羊水，母亲和胎儿的血液）多次，而不会破坏 怀孕本次重新提交的新的初步研究表明：1）通过 Rytvela至少7天（临床显著终点），2）测定Rytvela在大鼠中的药代动力学 模型，3）使用质谱法在血浆中定量Rytvela用于药代动力学研究的能力，以及3） 采用新技术（数字空间分析）研究母胎免疫反应 接口.在目标1中，我们将在NHP模型中确定Rytvela的药代动力学，并确定IL-1是否受到抑制 通过Rytvela延迟GBS诱导的早产的发生。目的2将确定Rytvela对IL-1的抑制是否可以 使用三种高通量多路复用技术改善胎盘、胎肺和脑的炎性损伤 分析：1）多维流式细胞术定量细胞群，2）数字空间分析， 询问离散组织（羊膜、绒毛膜、蜕膜）中免疫蛋白表达和途径活化 3）单细胞RNA-Seq用于胎盘绒毛膜羊膜的转录组学分析， 胎盘组织中单个细胞内的基因网络。这些实验代表了 在多个动物模型中进行的初步研究，并且对于确定IL-1是否是一种可行的分子是至关重要的。 预防早产和保护胎儿的目标。

PROJECT SUMMARY/ABSTRACT Preterm birth remains a significant cause of neonatal morbidity and mortality. Intra-amniotic infection and inflammation are important triggers for early preterm birth, which is associated with fetal injury mediated by cytokines and other inflammatory mediators in the amniotic fluid and fetus. The objective of this proposal is to establish the efficacy and safety of a novel selective allosteric interleukin-1 receptor (IL-1R) inhibitor (Rytvela) as an antenatal therapeutic strategy to prevent fetal injury and PTB. In pregnant mice, we have strong evidence that Rytvela is a potent suppressor of preterm birth and fetal injury induced by either lipopolysaccharide or lipotechoic acid. To enable translation of these discoveries to human pregnancy, we propose to test the efficacy and determine pharmacokinetics of Rytvela in a pregnant nonhuman primate model of preterm labor induced by Group B Streptococcus (GBS), a clinically important bacterium that colonizes the vagina and can cause preterm labor and neonatal sepsis. Our unique chronically catheterized nonhuman primate model has the greatest relevance to human pregnancy of any animal model and allows sampling in normally inaccessible compartments (amniotic fluid, maternal and fetal blood) multiple times without disrupting the pregnancy. New preliminary studies added to this resubmission demonstrate: 1) inhibition of preterm birth by Rytvela for at least 7 days (clinically significant endpoint), 2) determination of Rytvela pharmacokinetics in a rat model, 3) ability to quantify Rytvela using mass spectrometry in plasma for pharmacokinetics studies, and 3) incorporation of novel technology (Digital Spatial Profiling) to study immune responses at the maternal-fetal interface. In Aim 1, we will determine Rytvela pharmacokinetics in our NHP model and establish if IL-1 inhibition by Rytvela delays the onset of GBS-induced preterm labor. Aim 2 will determine if IL-1 inhibition by Rytvela can ameliorate inflammatory injury to the placenta, fetal lung and brain using three high-throughput multiplexed analyses: 1) multidimensional flow cytometry to quantitate cell populations, 2) Digital Spatial Profiling to interrogate immunologic protein expression and pathway activation in discrete tissues (amnion, chorion, decidua) of the placental chorioamniotic membranes, and 3) single cell RNA-Seq for transcriptomic profiling of regulatory gene networks within single cells in placental tissues. These experiments represent a natural progression of preliminary studies in multiple animal models and are essential to determining whether IL-1 is a viable molecular target for the prevention of preterm birth and fetal protection.