BSL3 Drug Screening Core

摘要(核心1) 高通量筛选(HTS)已成为现代药物发现的基石 学术界，允许实施大规模的筛查运动，其中数千人 化合物可以在寻找针对特定疾病的新药时进行测试。然而，压倒性的 现有的高温超导设施和专门设备大多建立在生物安全水平较低的实验室中。 (BSL)条件。这使得这种方法不可能对高致病微生物实施， 例如球孢子虫，因为处理这些病原体工作需要在高层内进行 生物控制设施。因此，该药物筛选核心的首要目标是建立独特的 在BSL-3条件下的能力，这将允许实施快速、高效和经济的 高温超导技术大规模筛选化学文库鉴定新抗球虫类化合物 探员们。更具体地说，我们建议：i)开发基于低密度和高密度微滴定板的模型 球藻(Coccidioidesspp.)在我们的BSL-3设施内进行符合HTS的敏感性测试，以及ii)充分开发一种 基于成像流式细胞术(IFC)的真菌细胞学分析(FCP)系统及其在药物中的应用 筛选和确定新药的作用模式。在CCRC内部，此核心将支持 项目1中的筛查和真菌细胞学分析活动，为TOP的下选提供服务 项目2和支持项目3检查免疫治疗活动的体内评估候选人 抗真菌药物和候选疫苗的疗效。此外，一旦完全确立，技术上的 这一核心的能力将提供给球虫科学界以及其他人 在医学真菌学领域，以及可能与BSL-3生物合作的其他研究人员。

Summary (Core 1) High-throughput screening (HTS) has become the cornerstone of modern drug discovery both in Industry and Academia, allowing for the implementation of large-scale screening campaigns where thousands of compounds can be tested in search for novel drugs against a given disease. However, the overwhelming majority of existing HTS facilities and specialized equipment are set up in laboratories under low biosafety level (BSL) conditions. This makes it impossible to implement this approach for highly pathogenic microorganisms, such as Coccidioides spp., since work with these pathogens needs to be conducted inside high level biocontainment facilities. Thus, the overarching goal of this Drug Screening Core is to establish unique capabilities within BSL-3 conditions, which will allow for the implementation of fast, efficient and economical HTS techniques for large-scale screening of chemical libraries for the identification of novel anti-coccidioidal agents. More specifically, we propose to: i) develop low- and high-density microtiter plate-based models for Coccidioides spp. susceptibility testing amenable to HTS inside our BSL-3 facility, and ii) to fully develop a novel fungal cytological profiling (FCP) system based on imaging flow cytometry (IFC), and adapt it for drug screening and to determine the mode of action of novel drugs. Within the CCRC, this Core will support the screening and fungal cytological profiling activities in Project 1, provide services for down-selection of top candidates for in vivo evaluation in Project 2, and support Project 3 activities examining immunotherapeutic efficacy of antifungal drugs and vaccine candidates. Furthermore, once fully established, the technical capabilities of this Core will be provided to the Coccidioides scientific community at large, as well as to others in the field of Medical Mycology, and potentially other investigators working with BSL-3 organisms.