Multi-omics of murine respiratory melioidosis

项目摘要 类鼻疽是一种致命的感染，通常由吸入、接种或摄入革兰氏阴性杆菌引起。 兼性胞内病原体和1级选择剂类鼻疽伯克霍尔德氏菌（Bps）。BPS最近 从美国南部的土壤中分离出来。全世界有16.5万例类鼻疽， 估计每年发生; 85，000（52%）这些患者死亡。肺炎存在于超过50%的 类鼻疽病例和死亡风险增加一倍以上。然而，为了开发新的靶向治疗方法， 需要更深入地了解宿主防御的肺和全身机制。我们的团队 结合了人类和实验类鼻疽病，肺宿主防御，败血症， 生物信息学我们已经建立了一个强大的Bps肺炎小鼠模型， 疾病表型同时，我们对一个大的前瞻性研究进行了无偏的多组学分析。 泰国东北部感染住院患者队列，目的是分类类鼻疽病例， 了解宿主对Bps的反应我们确定了不同的转录和代谢谱 与其他感染患者相比，与类鼻疽相关，并在每个组学中建立了强大的分类器 预测人类类鼻疽死亡。然而，为了全面研究机械 基础需要一个易于处理的实验模型，与人类疾病具有足够的可比性。 此外，为了研究呼吸性类鼻疽的致死性，需要对肺组织取样。我们假设 将比较多组学方法应用于患有呼吸性类鼻疽病的小鼠和人类， 对肺部宿主防御机制的重要见解，这些机制未能遏制感染并有助于 严重的结果和B）建立实验鼠模型与人感染的可比性。我们 提出以下具体目标：（1）。定义系统性宿主的多组学特征的时间轨迹 小鼠呼吸道类鼻疽的防御并识别代表宿主成功或失败的扰动 防御2)定义小鼠呼吸道类鼻疽中肺细胞特异性转录组学变化并鉴定 与肺宿主防御成功或失败相关的信号。3)识别共享的多组学 鼠和人呼吸道类鼻疽之间的特征。这些研究的结果将产生丰富的 关于全身和肺部宿主对鼠呼吸道类鼻疽反应的数据概要， 为失败的异质性和关键生物学途径提供了新的和全面的见解。 类鼻疽肺炎的宿主反应表型将这些发现与现有的人类 类鼻疽病数据将有助于确定进一步研究的临床相关目标，同时 提供了关于动物模型在再现人类中的优点和局限性的基本信息， 在多组学水平上的感染。

PROJECT SUMMARY Melioidosis is an often-fatal infection caused by inhalation, inoculation, or ingestion of the Gram-negative facultative intracellular pathogen and Tier 1 select agent Burkholderia pseudomallei (Bps). Bps has recently been isolated from the soil in the southern United States. Worldwide, 165,000 cases of melioidosis are estimated to occur each year; 85,000 (52%) of these patients die. Pneumonia is present in over 50% of melioidosis cases and more than doubles the risk of death. Yet, to develop novel, targeted therapeutics necessitates a deeper understanding of pulmonary and systemic mechanisms of host defense. Our team combines expertise in human and experimental melioidosis, pulmonary host defense, sepsis, and bioinformatics. We have developed a robust murine model of Bps pneumonia displaying mild and severe disease phenotypes. In parallel we have performed unbiased multi-omics analyses on a large prospective cohort of hospitalized patients with infection in NE Thailand with the goal of classifying melioidosis cases and understanding the host response to Bps. We identified distinct transcriptional and metabolomic profiles associated with melioidosis compared to other infected patients, and have built robust classifiers in each omics domain to predict death in human melioidosis. However, to comprehensively investigate mechanistic underpinnings requires a tractable experimental model with sufficient comparability to human disease. Moreover, to study the lethality of respiratory melioidosis requires sampling of lung tissue. We hypothesize that applying a comparative multi-omic approach to mice and humans with respiratory melioidosis will both a) yield critical insights into the pulmonary host defense mechanisms that fail to contain the infection and contribute to severe outcomes and b) establish comparability of the experimental murine model with human infection. We submit the following specific aims: 1). Define the temporal trajectory of multi-omic features of systemic host defense in murine respiratory melioidosis and identify perturbations representing success or failure of host defense. 2) Define lung cell-specific transcriptomic changes in murine respiratory melioidosis and identify signals that are associated with success or failure of pulmonary host defense. 3) Identify shared multi-omic signatures between murine and human respiratory melioidosis. The results of these studies will generate a rich compendium of data about the systemic and pulmonary host response to murine respiratory melioidosis and provide novel and comprehensive insights into the heterogeneity and key biological pathways underlying failed host response phenotypes of melioidosis pneumonia. Intersecting these findings with existing human melioidosis data will help to define clinically relevant targets for further investigation while simultaneously providing essential information about advantages and limitations of the animal model in recapitulating human infection at the multi-omic level.