Multi-organ omic model for sepsis therapeutic development

摘要 脓毒症-3被定义为威胁生命的器官功能障碍，由机体对 一种感染。脓毒症的早期特征是内皮细胞的异常激活，这启动了一种 通过释放各种介质导致炎症信号事件的级联，导致白细胞黏附， 如果不加控制，会导致迁移、组织损伤和多器官功能障碍综合症(MODS)。到目前为止， 脓毒症的治疗方法在很大程度上是支持性的，但没有具体的方法。 可用于保护内皮细胞功能障碍的药物治疗。最近所有的败血症药物 在啮齿动物身上开发的药物在临床试验中都失败了，这在很大程度上是因为物种和 不同表型的内皮细胞在功能、形态和基因组上表现出异质性 表情模式。因此，需要利用组学最新发展的新方法来 研究这种异质性如何影响脓毒症的治疗反应。在这个项目中，我将聘用OMIC 并在硅胶模型中研究特定蛋白靶标在脓毒症进展中的作用。我们的团队已经 确定蛋白激酶C-增量(PKCδ)是炎症反应的关键调节因子，我将使用我的 确定蛋白激酶Cδ在人肺、肝和肾脏炎症信号转导中作用的模型 脓毒症的血管内皮细胞。总体而言，这些研究将有助于确定可用药/治疗靶点，然后 使用我们的新型微生理学分析(MPA)进行了实验验证。我假设我的求婚 在我们的MPA中，全面的蛋白质组学模型和相应的验证实验将 对蛋白质靶标在预测人类生理反应中的作用提供独特的见解 在败血症和正常情况下。我们的长期目标是开发一种方法来合理地设计 治疗败血症的治疗学。我的研究的具体目标是：1)创造一个全面的，在计算机上 预测败血症和正常情况下人类生理反应的蛋白质组学模型和2)用途 一种新的MPA来验证目标1中预测的蛋白质靶标的作用。这些协同研究将集中在 内皮细胞异质性在脓毒症中的作用以及蛋白激酶Cδ和其他蛋白靶标在脓毒症中的作用 人感染脓毒症模型和微血管内皮细胞反应的调控 脓毒症患者和健康受试者的内皮细胞、白细胞和血浆增加可译性 在治疗发展方面。

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.