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

吸入烟曲霉孢子每年在世界范围内造成300,000,000种慢性和200,000种侵袭性疾病。侵袭性曲霉病的总死亡率为50%，但如果诊断延迟或漏诊，死亡率接近100%，而在某些患者队列中，死亡率为75%。此外，目前感染耐唑菌株的死亡率接近100%，自2006年以来增加了40%。因此，迫切需要开发新的策略来临床治疗真菌感染。Bertuzzi团队之前的工作已经证明，呼吸道上皮细胞(AECs)在摄取时有效地杀死烟曲霉菌孢子，并且这一过程在慢性阻塞性肺疾病(COPD)患者中发生改变，COPD是众所周知的致衰性真菌肺部疾病的危险因素。因此，他们工作的首要假设是，AEC对吸入烟曲霉孢子的健康清除至关重要，保护性AEC反应的失调是曲霉相关疾病的有力驱动因素。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烟曲霉孢子清除以及伴随的呼吸道炎症反应的动态平衡的基础上，定义转录和免疫学反应，以确定健康的AEC如何识别和杀死烟曲霉菌，以及这一过程如何影响免疫反应和疾病中的失调调节。为此，我们建议在体外、体外和健康供者和COPD患者的原代AEC中对烟曲霉菌与AEC的相互作用进行第一次现场转录和免疫学分析。总体目标是从机制上了解呼吸道上皮的抗真菌效力及其在疾病中的功能障碍，本研究最终旨在为免疫调节剂的识别提供信息，以促进这些和其他致命性呼吸道感染的临床治疗，并最终减轻它们对人类的疾病负担。

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.