Discovery of Novel Therapeutics for Fungal Pathogens

发现治疗真菌病原体的新疗法 摘要 真菌中枢神经系统(CNS)感染仍然是导致死亡和发病的重要原因。 新生隐球菌和白色念珠菌是重要的真菌。 引起中枢神经系统感染的病原体。成功治疗新生葡萄球菌和白色念珠菌中枢神经系统感染 由于抗菌素耐药性的发展和有限的抗真菌药物库而受到损害 耐药的新生葡萄球菌和白色念珠菌感染。本申请书是根据RFA-AI-15-054号文件提交的， 建议开展以小说发现和发展为重点的早期翻译研究 治疗真核真菌病原体、新生葡萄球菌和白色念珠菌引起的中枢神经系统感染。 最近的研究表明，感染中枢神经系统的病原体具有穿透血脑屏障的能力， 中枢神经系统感染发生发展的关键步骤。自从新生念珠菌和白色念珠菌传播到 大脑很可能涉及其穿透血脑屏障，我们开发了体外血脑 用人脑微血管内皮细胞(HBMEC)建立屏障模型，研究新生葡萄球菌和C. 白念珠菌穿透血脑屏障。在胶原涂层上培养的HBMEC单层 Transwell插件，表现出紧密连接形成和极化的形态和功能特性 单层，血脑屏障内皮细胞的一种独特属性。我们发现新生隐孢子虫和 白念珠菌菌株表现出穿透血脑屏障的能力。我们假设目标是C。 新生杆菌和白色念珠菌对血脑屏障的渗透提供了一种新的方法来发现 治疗真菌中枢神经系统感染的新药物。这一假说得到了我们初步确定的 孟鲁司特(半胱氨酰白三烯1型受体拮抗剂或CysLT1)和吉非替尼(吉非替尼) 表皮生长因子受体(EGFR)抑制新生葡萄球菌和白色念珠菌对皮肤的穿透 血脑屏障。CysLT1和EGFR以前没有被认为是对真菌的贡献 是治疗新生葡萄球菌和白色念珠菌中枢神经系统感染的新靶点。 CysLT1和EGFR对新生念珠菌和白色念珠菌贡献的测定和验证 因此，血脑屏障的穿透将有助于发现新的宿主特异性治疗方法。 新生念珠菌和白色念珠菌共有的靶点，与RFA-AI-15-054的FOA一致。

Discovery of novel therapeutics for fungal pathogens Abstract Fungal central nervous system (CNS) infection continues be an important cause of mortality and morbidity. Cryptococcus neoformans (C. neoformans) and Candida albicans (C. albicans) represent the important fungal pathogens causing CNS infection. Successful treatment of C. neoformans and C. albicans CNS infection is compromised by development of antimicrobial resistance and limited repertoire of antifungal drugs against resistant C. neoformans and C. albicans infection. This application, submitted in response to RFA-AI-15-054, proposes to develop early stage translational research focused on the discovery and development of novel therapeutics against CNS infection caused by eukaryotic fungal pathogens, C. neoformans and C. albicans. Recent studies have shown that CNS-infecting pathogens exhibit the ability to penetrate the blood-brain barrier, the essential step in the development of CNS infection. Since C. neoformans and C. albicans dissemination to the brain is likely to involve their penetration of the blood-brain barrier, we developed the in vitro blood-brain barrier model with human brain microvascular endothelial cells (HBMEC) to investigate C. neoformans and C. albicans penetration of the blood-brain barrier. The HBMEC monolayer, upon cultivation on collagen-coated Transwell inserts, exhibits morphological and functional properties of tight junction formation and a polarized monolayer, a unique property of the blood-brain barrier endothelial cells. We showed that C. neoformans and C. albicans strains exhibited the ability to penetrate the blood-brain barrier. We hypothesize that targeting C. neoformans and C. albicans penetration of the blood-brain barrier provides a novel approach for discovery of novel therapeutics for fungal CNS infection. This hypothesis is supported by our preliminary identification of montelukast (an antagonist of cysteinyl leukotriene type 1 receptor or CysLT1) and gefitinib (an inhibitor of epidermal growth factor receptor or EGFR) that inhibited C. neoformans and C. albicans penetration of the blood-brain barrier. CysLT1 and EGFR have not been previously recognized for their contribution to fungal CNS infection and represent novel therapeutic targets for C. neoformans and C. albicans CNS infection. Determination and validation of CysLT1 and EGFR for their contribution to C. neoformans and C. albicans penetration of the blood-brain barrier will, therefore, facilitates discovery of novel host-specific therapeutic targets common to C. neoformans and C. albicans, which is consistent with the FOA of RFA-AI-15-054.