Development and application of innovative tools to mitigate White Nose Syndrome, a lethal fungal disease decimating North American bat populations

该项目的重点是开发一种基于免疫的策略来预防白鼻综合征（WNS），这是一种致命的真菌感染，在冬眠的蝙蝠中传播，正在北美蔓延。这项研究将加深对真菌病原体如何攻击的理解，并指导保护蝙蝠免受WNS破坏性影响的管理策略。具体来说，研究小组将研究真菌病原体是如何通过蝙蝠表皮生长因子受体的作用侵入皮肤的，这可能会对哺乳动物的这一过程有更广泛的了解。此外，该团队将尝试一种将疫苗接种与局部阻断感染的药物相结合的新方法，其目标是促进几种蝙蝠物种的保护，维持生物多样性和支持生态系统健康。此外，该项目以培训和教学早期职业科学家和野生动物生物学学员为特色，并基于学术机构和联邦机构-美国地质勘探局和美国野生动物管理局的合作。本项目的基础研究是进一步了解真菌Pseudogymnoascus destructans（简称Pd）是如何通过附着在蝙蝠皮肤上引发感染的。已知Pd与角质形成细胞（如上皮细胞）接触。该项目利用了首个来自lucifugus的角质细胞细胞系，这是一种对WNS高度敏感的蝙蝠物种。角化细胞细胞系为研究Pd如何侵入蝙蝠皮肤并引发感染提供了理想的模型。初步数据显示：(i)表皮生长因子受体（EGFR）受体的特性，介导Pd对角质形成细胞的“隐形”侵袭；（ii） FDA批准的药物吉非替尼（gefitinib）的效用，阻断Pd对细胞的侵袭。实验旨在通过CRISPR编辑角质形成细胞正式测试EGFR在Pd侵袭中的作用。由于易感和耐药蝙蝠的EGFR结构不同，来自耐药蝙蝠的EGFR将用于补充敲除细胞系，以评估EGFR在解释WNS易感性中的作用。保护生物学是这项工作的一个重要方面。WNS的预防战略将与美国地质调查局在国家野生动物健康中心与美国鱼类和野生动物管理局合作实施。因此，将在美国几个州进行实地研究，以调查吉非替尼与这些研究人员开发的预防Pd感染的保护性疫苗的使用情况。该项目由环境生物学和综合有机体系统部以及保罗·g·艾伦家族基金会共同资助。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

This project focuses on developing an immune-based strategy to prevent White Nose Syndrome (WNS), a lethal fungal infection of hibernating bats that is spreading across North America. This research will develop a deeper understanding of how the fungal pathogen attacks, and guide management strategies for protecting bats against the devastating impact of WNS. Specifically, the research team will interrogate how a fungal pathogen invades skin through the role of the epidermal growth factor receptor in bats, possibly resulting in a broader understanding of such processes in mammals in general. Additionally, the team will attempt a novel approach of combining vaccination with topical drugs that block infection – with the goals to advance the conservation of several bat species, maintain biodiversity and support ecosystem health. In addition, this project features the training and teaching of early career scientists and wildlife biology trainees and is based on the collaboration of an academic institution and federal agencies – USGS and USFWS.The fundamental research associated with this project is to further understand how the fungus Pseudogymnoascus destructans (Pd for short) initiates infection by adhering to bat skin. It is known that Pd makes contact with keratinocytes (e.g. epithelial cells). This project makes use of a first-of-its-kind keratinocyte cell line from Myotis lucifugus, a bat species that is highly susceptible to WNS. The keratinocyte cell line has provided an ideal model to study how Pd invades bat skin and initiates infection. Preliminary data reveal: (i) the identity of a receptor – epidermal growth factor receptor (EGFR) – that mediates keratinocyte “stealth” invasion by Pd and (ii) the utility of the FDA approved drug gefitinib® that blocks Pd invasion of the cells. Experiments are designed to formally test the role of EGFR in Pd invasion via CRISPR editing of keratinocytes. Because EGFR differs in its structure in susceptible and resistant bats, EGFR from resistant bats will be used to complement the knockout cell line to assess the role of EGFR in explaining WNS susceptibility. Conservation biology is an important aspect of the work. WNS prevention strategies will be implemented in collaboration with the U.S. Geological Survey at the National Wildlife Health Center in partnership with the U.S. Fish and Wildlife Service. Field studies will thus be performed in several U.S. states to investigate the use of gefitinib together with a protective vaccine created by these investigators against Pd infection.This project is being supported via a joint program involving the Divisions of Environmental Biology and Integrative Organismal Systems and the Paul G. Allen Family Foundation.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.