Defining the single-cell transcriptional and immunological atlas driving antifungal mucosal responses

烟曲霉孢子的吸入每年在全世界引起> 3，000，000例慢性疾病和> 200，000例侵袭性疾病。侵袭性曲霉菌病的总体死亡率为50%，但如果诊断延迟或漏诊，死亡率接近100%，并且在某些患者队列中死亡率>75%。此外，目前感染唑类耐药分离株的死亡率接近100%，自2006年以来增加了40%。Bertuzzi小组先前的工作已经证明气道上皮细胞（AECs）可以有效地杀死A.研究人员发现，烟曲霉孢子在摄取后会被吸入体内，并且这一过程在患有慢性阻塞性肺病（COPD）的患者中发生改变，COPD是使人衰弱的真菌性肺病的一个众所周知的风险因素。因此，他们工作的首要假设是AEC对吸入A的健康清除至关重要。烟曲霉孢子和保护性AEC反应失调是曲霉相关疾病的潜在驱动因素。利用最先进的单细胞技术，我们现在希望在有效和功能失调的清除A的基础上定义转录和免疫应答。烟曲霉孢子和伴随的气道炎症反应的稳态，以确定健康的AEC如何识别和杀死A。以及这个过程如何影响免疫反应和在疾病中失调。为此，我们建议进行首次在现场转录和免疫分析的A。根据疾病严重程度分层的健康供体和COPD患者的体外、离体和原发性AEC中的烟曲霉-AEC相互作用。总体目标是获得对气道上皮抗真菌效力及其疾病功能障碍的机制理解，这项研究的最终目的是为免疫识别提供信息，调节剂，以促进这些和其他致命呼吸道感染的临床管理，并最终减少其在人类中的疾病负担。

Inhalation of Aspergillus fumigatus spores cause >3,000,000 chronic and >200,000 invasive diseases annually worldwide. Invasive aspergillosis carries a 50% mortality rate overall, but mortality rate approaches 100% if diagnosis is delayed or missed and it is >75% in certain cohorts of patients. Additionally, current mortality approaches 100% for infections with azole-resistant isolates, which have increased by 40% since 2006. Thus, there is an urgent need for the development of novel strategies for the clinical management of fungal infection.Previous work from the Bertuzzi's group has demonstrated that airway epithelial cells (AECs) efficiently kill A. fumigatus spores upon uptake and that this process is altered in patients with chronic obstructive pulmonary disease (COPD), a well-known risk factor for debilitating fungal lung disease. Therefore, the overarching hypothesis of their work is that AECs critically contribute to healthy clearance of inhaled A. fumigatus spores and that dysregulation of protective AEC responses represents a potent driver of aspergillus-related diseases. Using state-of-the-art single-cell technologies, we now wish to define the transcriptional and immunological responses at the base of effective and dysfunctional clearance of A. fumigatus spores and concomitant homeostasis of airway inflammatory responses, to determine how healthy AECs recognize and kill A. fumigatus and how this process influences immune responses and is dysregulated in disease. To this end, we propose to perform first-in-field transcriptional and immunological analyses of the A. fumigatus-AEC interaction in vitro, ex vivo and in primary AECs from healthy donors and COPD patients stratified according to disease severity.With the overall aim of obtaining a mechanistic understanding of the antifungal potency of the airway epithelium and its dysfunction in disease, this research ultimately aims to inform the identification of immune-modulators to facilitate clinical management of these and other lethal respiratory infections and ultimately reduce their disease burden in humans.