Targeted Inhibition of Interleukin-1 beta to Prevent Preterm Birth

项目摘要/摘要 早产仍然是新生儿发病率和死亡率的重要原因。羊膜内感染和 炎症是早期早产的重要诱因，通过以下途径与胎儿损伤有关 羊水和胎儿中的细胞因子和其他炎症介质。这项提议的目标是 一种新型选择性变构白介素1受体(IL-1R)抑制剂的有效性和安全性 (Rytvela)作为预防胎儿损伤和肺结核的产前治疗策略。在怀孕的小鼠身上，我们有 强有力的证据表明，Rytvela是一种有效的抑制早产和胎儿损伤的药物 脂多糖或脂磷壁酸。为了能够将这些发现转化为人类怀孕，我们 建议在怀孕的非人灵长类动物模型上测试Rytvela的疗效并确定其药代动力学 B组链球菌(GBS)是一种临床上重要的细菌，在 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 在所有动物模型中，模型与人类怀孕的相关性最大，并允许正常采样 多次接触不到的隔室(羊水、母体和胎儿血液)，而不会破坏 怀孕了。重新提交的新的初步研究表明：1)通过以下方式抑制早产 Rytvela至少7天(临床显著终点)，2)Rytvela在大鼠体内的药代动力学测定 模型，3)在药物动力学研究中使用血浆中的质谱学来定量Rytvela的能力，以及3) 结合新技术(数字空间剖面图)研究母婴免疫反应 界面。在目标1中，我们将确定Rytvela在我们的NHP模型中的药代动力学，并确定是否抑制IL-1 通过Rytvela延缓GBS诱导的早产的开始。Aim 2将确定Rytvela是否可以抑制IL-1 三种高通量复合体减轻胎盘、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)数字空间剖面图 询问免疫蛋白在离散组织(羊膜、绒毛、蜕膜)中的表达和途径激活 以及3)单细胞RNA-Seq用于转录图谱的调控 胎盘组织中单个细胞内的基因网络。这些实验代表了一个自然发展的过程 在多种动物模型中的初步研究，对于确定IL-1是否是一个可行的分子是必不可少的 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.