Humanized lung implant mouse model to study Staphylococcus aureus airway interactions

总结 金黄色葡萄球菌是一种重要的革兰氏阳性人类病原体， 感染了包括肺部在内的多个身体部位它会导致严重的发病率和死亡率， 具有较高的抗生素耐药性发生率，并且是被确定为需要增加的病原体。 研究发现新的疗法。我们研究的长期目标是 了解宿主-病原体之间的相互作用S.金黄色葡萄球菌和宿主免疫系统。是 希望对这种相互作用的更好理解可以导致新的治疗靶向 细菌或调节宿主以阻止这种多药耐药病原体。客观 这项建议的目的是表征人类肺部的先天免疫反应以及 S.金黄色葡萄球菌的气道，并确定遗传因素的重要性， 感染这种方法的基本原理是，需要制定新的目标， 抗生素/抑制剂或疫苗。金黄色。这是由于其高水平的抗生素 在小鼠模型中的耐药性和先前有希望的疫苗候选物未能 转化为有效的人类疫苗。我们的初步数据显示， 为S.金黄色葡萄球菌感染在肺部，让作用 人类特异性毒力因子有待研究和了解的免疫反应 感染我们提出了一种新的人源化小鼠模型， 人类肺组织我们还展示了使用Tn-seq文库来鉴定新的 呼吸道感染的重要因素。这一贡献是重要的，因为它将提供一个 了解人类免疫系统，包括上皮细胞如何对S. aureus，以及S.金黄色葡萄球菌对这种环境和它所采用的因素作出反应， 持续感染。本研究的创新之处在于采用了一种新的感染模型 利用我们在感染的人性化模型上的经验。该模型包含 人类肺组织来更好地复制人类肺。我们将描述这种反应， 在转录、蛋白质和细胞水平上的多个水平。全球理解方法 病原体反应、RNA-seq和Tn-seq将被组合以鉴定新的遗传元件 在发病机制中起重要作用。鉴于目前疫苗和现有药物管道的失败， 在细胞特征毒力以外的人类模型背景下研究基因产物 这些因素可能会导致新的发现。

Summary Staphylococcus aureus in an important Gram positive human pathogen that is capable of infecting numerous bodily sites including the lung. It leads to significant morbidity and mortality, has a high incidence of antibiotic resistance and is a pathogen identified as needing increased research to discovery new therapeutics. The long term goal of our research is to better understand the host-pathogen interaction between S. aureus and the host immune system. It is hoped that an improved understanding of this interaction could lead to novel therapies targeting the bacterium or modulating the host to thwart this multidrug resistant pathogen. The objective of this proposal is to characterize the innate immune response of the human lung as well as the initial response of S. aureus to the airway and identify the genetic factors important for causing infection. The rationale for this approach is that new targets are needed to either develop antibiotics/inhibitors or vaccines against S. aureus. This is due to its high level of antibiotic resistance and previously promising vaccine candidates in murine models have failed to translate to efficacious human vaccines. Our preliminary data shows that humanized mice represent an improved model for the study of S. aureus infection in the lung, allowing the role of human specific virulence factors to be studied and understand the immune response to infection. We present a novel model of humanized mice whereby mice are implanted with human lung tissue. We have also demonstrated the use of a Tn-seq library to identify new factors important for infection in the airway. This contribution is significant as it will provide an understanding of how the human immune system, including the epithelium responds to S. aureus, as well as how S. aureus reacts to this environment and the factors that it employs to sustain infection. The innovation of this research is the utilization of a new model of infection that leverages our experience with humanized models of infection. This model incorporates human lung tissue to better replicate the human lung. We will characterize this response at multiple levels, at the transcriptional, protein and cellular level. Global approaches to understand the pathogen response, RNA-seq and Tn-seq will be combined to identify new genetic elements important in pathogenesis. Given present failure with vaccines and current drug pipelines, investigating gene products in the context of human model outside of ell-characterized virulence factors might lead to novel discoveries.