Multi-organ omic model for sepsis therapeutic development

摘要 脓毒症-3被定义为由身体对脓毒症的宿主反应失调引起的危及生命的器官功能障碍。 感染脓毒症的早期特征是内皮细胞的活化失调，其启动了一种新的免疫调节机制。 通过释放各种介质，导致白细胞粘附， 迁移、组织损伤和多器官功能障碍综合征（MODS）。到目前为止， 治疗脓毒症的治疗方法在很大程度上是支持性的，但没有特异性的 现有的保护内皮细胞功能障碍的药物疗法。最近所有的败血症药物 在啮齿类动物中开发的药物在临床试验中失败了，这在很大程度上是因为物种和 内皮细胞的不同表型在功能、形态和组学方面表现出异质性 表达模式因此，需要利用组学的最新发展的新方法， 研究这种异质性如何影响脓毒症治疗的反应。在这个项目中，我将采用omic 和计算机模拟模型，以研究特定蛋白质靶点在脓毒症进展中的作用。我们集团 我发现蛋白激酶C-δ（PKCδ）是炎症反应的关键调节因子，我将用我的 模型以确定PKCδ在人肺、肝和肾中炎症信号传导进展中的作用 内皮细胞在脓毒症中的作用总体而言，这些研究将有助于确定可药物化/治疗目标，然后将 实验验证使用我们的新的微生理测定（MPA）。我假设我的提议 全面的，在计算机蛋白质组学模型和相应的验证实验，在我们的MPA将 对蛋白质靶点在预测人类生理反应中的作用提供了独特的见解 在败血症和正常情况下。我们的长期目标是开发一种方法， 用于治疗脓毒症的治疗剂。我的研究的具体目标是1）创建一个全面的，在硅片 蛋白质组学模型，以预测人类在脓毒症和正常条件下的生理反应，以及2）使用 一种新的MPA，以验证目标1中预测的蛋白质靶点的作用。这些协同研究将侧重于 内皮细胞异质性在脓毒症中的作用，以及PKCδ和其他蛋白靶点在脓毒症中的作用。 使用计算机模拟模型和使用人MPA调节脓毒症中的内皮细胞应答 内皮细胞以及来自脓毒症患者和健康受试者的白细胞和血浆，以增加可翻译性 在治疗发展中。

ABSTRACT Sepsis-3 is defined as life-threatening organ dysfunction caused by the body’s dysregulated host response to an infection. An early feature of sepsis is the dysregulated activation of endothelial cells, which initiates a cascade of inflammatory signaling events by releasing various mediators, leading to leukocyte adhesion, migration, tissue damage and multiple organ dysfunction syndrome (MODS) if left uncontrolled. To date, therapeutic approaches for the treatment of sepsis are largely supportive, but there are no specific pharmacologic therapies available that protect from endothelial cell dysfunction. All sepsis drugs recently developed in rodents have failed in clinical trials, in large part because of the differences in species and the diverse phenotypes of endothelial cells demonstrating heterogeneity in function, morphology and omic expression patterns. Novel methods leveraging recent developments in omics are therefore needed to investigate how this heterogeneity impacts response to therapeutics in sepsis. In this project, I will employ omic and in silico models to investigate the role of specific protein targets in sepsis progression. Our group has identified Protein Kinase C-Delta (PKCδ) as a critical regulator of the inflammatory response, and I will use my model to determine the role of PKCδ in the progression of inflammatory signaling in human lung, liver and kidney endothelium in sepsis. Overall, these studies will help identify druggable/therapeutic targets that will then be experimentally validated using our novel microphysiological assay (MPA). I hypothesize that my proposed comprehensive, in silico proteomics model and corresponding validation experiments in our MPA will provide unique insight on the role of protein targets in predicting physiological responses in humans under septic and normal conditions. Our long-term goal is to develop a methodology to rationally design therapeutics for treating sepsis. The specific aims of my study are to 1) Create a comprehensive, in silico proteomics model to predict physiological responses in humans under septic and normal conditions and 2) Use a novel MPA to validate the role of the protein targets as predicted in Aim 1. These synergistic studies will focus on the role of the endothelial cell heterogeneity in sepsis, and the role of PKCδ and other protein targets in regulating the endothelial cell response in sepsis using an in silico model and an MPA employing human endothelial cells, and leukocytes and plasma from septic patients and healthy subjects to increase translatability in therapeutic development.