Exploiting interspecies/interkingdom communication for the discovery and development of novel natural products antifungal therapeutics

总结 念珠菌病是一种机会性真菌感染， 是早产儿新生儿血流感染的常见原因。疟疾是一种 是新生儿重症监护病房真菌感染相关死亡率的主要原因。白色念珠菌是 引起口腔、皮肤、生殖器和泌尿道感染的主要真菌，以及全身或 多器官受累的播散性疾病。与下列疾病有关的粗死亡率和可归因死亡率 念珠菌病和侵袭性念珠菌感染非常高。尽管进行了抗真菌治疗， 侵袭性念珠菌病死亡，近60%的幸存者发生神经发育障碍。给定 合适和有效的抗真菌药物数量有限，加上病原体的耐药性增加 唑类抗真菌剂的发展最终导致对新抗真菌剂的需求。治疗剂的开发， 具有抑制生物膜形成和调节炎症反应的能力， 念珠菌病的防治。 在题为“真核病原体新型治疗药物的发现/开发”的R21/R33 RFA中， 建议发现和开发基于天然产物的药剂作为新型抗真菌治疗剂。我们将利用 通过培养白色念珠菌与来自大的、充分表征的个体生物体的共培养策略， 细菌和真菌的集合。假设是通过模仿两个人之间的自然竞争 生物体，我们将能够触发沉默的次级代谢途径的生物合成， 能抑制C.白色念珠菌生长。由于在最初的R21期间进行了大量的共培养实验， 阶段，我们建议使用最近开发的微流控装置来增加这些共培养物的产量 用于鉴定先导化合物的实验。在项目的R33阶段，我们建议优化顶部 通过工程化天然产物的生物合成途径，从该项目的R21阶段中获得成功。的 这项工作的转化意义在于，它可能导致一种新的药理学治疗，以预防或 治疗人体念珠菌病。

SUMMARY Candidiasis is an opportunistic fungal infection that has high prevalence among immunocompromised individuals and is a common cause of neonatal bloodstream infections in premature infants. Candidiasis is a leading cause of fungal infections-related mortality in the neonatal intensive care unit. Candida albicans is the major fungal organism responsible for oral, cutaneous, genital and urinary tract infections, and systemic or disseminated disease with multi-organ involvement. The crude and attributable mortality rates associated with candidiasis and invasive candidal infections are very high. Despite antifungal treatment, 20% of infants who develop invasive candidiasis die, and neurodevelopmental impairment occurs in nearly 60% of survivors. Given the limited number of suitable and effective antifungal drugs, together with increasing resistance of the pathogens to azole antifungal agents culminate in the need for new antifungals. The development of therapeutic agents that have the ability to inhibit the biofilm formation and modulate inflammatory response can have major impact on the prevention and treatment of candidiasis. In this R21/R33 RFA titled “Discovery/Development of Novel Therapeutics for Eukaryotic Pathogens” we propose to discover and develop natural product based agents as novel antifungal therapeutics. We will utilize a coculture strategy by culturing Candida albicans with individual organisms from large, well characterized bacterial and fungal collections. The hypothesis is that by mimicking the natural competition between two organisms we will be able to trigger the biosynthesis of the silent secondary metabolism pathways that will be able to inhibit C. albicans growth. Because of the large number of coculture experiments during the initial R21 stage, we propose to use a recently developed microfluidic device to increase the output of these coculture experiments for the identification of lead compounds. In the R33 stage of the project we propose to optimize top hits from the R21 phase of the project by engineering the biosynthetic pathways of the natural products. The translational significance of this work is that it could lead to a novel pharmacological therapeutic to prevent or treat candidiasis in humans.