Characterization of Neutrophil Decision making during Swarming to Fungal Pathogens

项目摘要 侵袭性真菌感染是严重的公共卫生威胁，并与高死亡率相关， 这表明目前的抗真菌治疗是不够的。虽然先天免疫是至关重要的 对于宿主对真菌的防御，这些复杂的宿主-病原体相互作用仍然不清楚。近日一 中性粒细胞是抗真菌防御的关键先天免疫细胞，其新的行为已经被描述， 中性粒细胞群集。群集被认为在遏制病原微生物方面发挥了作用，但其作用 在抗真菌防御中的特征性很差，代表了在中性粒细胞 功能推动这项研究应用的假设是，群集的早期事件是至关重要的 病原体是否将被成功遏制的决定因素，以及这些途径的特征将 突出了在感染和炎症期间优化中性粒细胞功能的新治疗选择。 不幸的是，对这些宿主-病原体群集相互作用的详细研究受到以下因素的阻碍： 目前实验分析的缺点。为了解决这个问题，我们开发并优化了一种新的 微型设备，使我们能够表征人类中性粒细胞群集活真菌病原体。这 显微点样分析使我们能够在大阵列中对活微生物进行图案化， 和用于分子分析的上清液。因此，本申请的目的是利用新颖的 微尺度工具，以允许宿主免疫和活细胞之间的动态相互作用的严格调查， 真菌在集群中扩展我们对集群生物学的理解。我们将通过追求 两个具体目标：（1）确定中性粒细胞决定启动群集的分子机制 行为和（2）阐明增强群集介导的真菌杀伤的分子途径。短期 长期而言，这项研究有望产生关于中性粒细胞群集在抗真菌治疗中的作用的关键知识。 防御利用这些工具及其产生的知识，长期目标是开发 为侵袭性真菌感染患者提供改进和新颖的治疗选择。 这项工作也将提供一个坚实的基础，候选人可以用来实现他的直接职业生涯 实现一个独立的，终身教职的目标，并朝着他的长期职业生涯的进展 终身研究教师职位的目标，具有研究先天宿主的独特学术研究计划， 真菌感染期间的病原体相互作用。为了实现这些职业目标，候选人还将 组建一个有效的指导和咨询团队，以促进研究的成功完成， 不断提高实验室和实验室管理技能。这些职业目标和 因此，拟议的研究正在履行NIAID的使命，即追求和确定新的治疗策略， 对抗感染，并支持初级科学家通过K22过渡到独立的教师职位。

Project Summary Invasive fungal infections are a serious public health threat and are associated with high mortality rates, demonstrating that current antifungal therapies are inadequate. While innate immunity is known to be critical for host defense against fungi, these complex host-pathogen interactions remain poorly elucidated. Recently, a novel behavior of neutrophils, a key innate immune cell for antifungal defense, has been characterized, that of neutrophil swarming. Swarming is thought to play a role in the containment of pathogenic microbes, but its role in antifungal defense is poorly characterized, representing a significant gap in knowledge in neutrophil function. The hypothesis driving this research application is that the early events in swarming are critical determinants of if the pathogen will be successfully contained and that characterization of these pathways will highlight novel therapeutic options for optimizing neutrophil function during infection and inflammation. Unfortunately, detailed study of these host-pathogen swarming interactions has been hindered by the shortcomings of current experimental assays. To address this, we have developed and optimized a novel microscale device to allow us to characterize human neutrophil swarming to live fungal pathogens. This microspotting assay allows us to pattern live microorganisms in large arrays, with direct access both visually and to supernatants for molecular analysis. The objective of this application is therefore to leverage novel microscale tools to allow rigorous investigation of the dynamic interactions between host immunity and live fungi during swarming to expand our understanding of swarming biology. We will do this by via the pursuit of two specific aims (1) Identify the molecular mechanisms by which neutrophils decide to initiate swarming behavior and (2) Elucidate the molecular pathways that enhance swarming mediated fungal killing. In the short term, this research is expected to generate critical knowledge on the role of neutrophil swarming in antifungal defense. Leveraging these tools and the knowledge they generate, the long term goal is the development of improved and novel therapeutic options for patients with invasive fungal infections. The work will also provide a strong foundation which the candidate can use to achieve his immediate career goals of attaining an independent, tenure-track faculty position and to progress towards his long term career goals of a tenured research faculty position with a unique academic research program studying innate host- pathogen interactions during fungal infection. In order to attain these career goals, the candidate will also assemble an effective mentoring and consulting team to promote the successful completion of research and the continuing improvement of grantsmanship and lab management skills. These career goals and the proposed research are therefore fulfil NIAID’s mission to pursue and identify novel therapeutic strategies to combat infection and to support the transition of junior scientists into independent faculty positions via the K22.