Administrative Supplement: Borrelia gene products critical for natural infection cycle

项目总结/摘要 莱姆病是世界上许多地方最普遍的蜱传感染，包括美国，哪里 每年有超过45万个新病例。这种疾病是由一组细菌病原体， 广义burgdorferi，通过涉及硬蜱的复杂地方性感染循环在自然界中茁壮成长 肩胛蜱和各种脊椎动物宿主。尽管做了很多努力，但感染仍然难以控制， 主要原因是缺乏包括疫苗在内的预防战略， 感染，以及使用现有抗生素未能实现完全治愈。具体来说，几个月后， 在标准抗生素治疗的情况下，一部分患者可能会经历一系列持续性或复发性 症状，这被称为慢性莱姆病或治疗后莱姆病综合征（PTLDS）。 虽然细菌感染的持续性可能会或可能不会有助于病原学或发病机制， PTLDS，其治疗选择仍然未知。因此，疫苗和新药的开发是 非常有必要我们已经确定了一个关键的毒力决定因子，称为BbHtrA， 在宿主感染性和螺旋体持久性中的作用。蛋白质的缺失使得病原体不- 在哺乳动物宿主中传染。基于这些信息，我们提出了一种治疗策略， 通过小分子药物破坏蛋白质功能。为此，我们将鉴定出 通过采用设计用于大的BbHtrA蛋白酶的高通量筛选（HTS）的测定来破坏BbHtrA蛋白酶活性， 基于nanoDSF和荧光蛋白酶抑制技术的化合物库。点击此屏幕 将通过第二次细胞测定验证，揭示可渗透穿过B的有效化合物。 伯氏膜化合物筛选将与国家药物研究中心合作进行。 先进的转化科学（NCATS）研究人员，他们提供专业知识，基础设施和现代化的 化合物库，并在HTS和药物发现方面拥有丰富的经验。我们还将尝试 开展BbHtrA的结构研究，这对开发新的治疗药物很重要 靶向蛋白质。将在莱姆病的整个动物模型中验证选定的命中分子。 在未来更大的补助金申请中进行审查。该项目采用了尖端技术， 相互作用的毒力决定因素，以打击莱姆病，一种新的战略，可能会避免PTLDS。 此外，同样的方法可以作为对抗其他蜱传感染的范例，包括 这些疾病是由最近发现的更致命的莱姆病病原体引起的。

PROJECT SUMMARY/ABSTRACT Lyme disease is the most prevalent tick-borne infection in many parts of the world, including the U.S., where over 450,000 new cases occur annually. The disease is inflicted by a group of bacterial pathogens, Borrelia burgdorferi sensu lato, that thrive in nature through a complex enzootic infection cycle involving Ixodes scapularis ticks and a variety of vertebrate hosts. Despite much effort, the infection remains difficult to control, largely due to the absence of preventive strategies including vaccines, difficulties in the diagnosis of early infection, and failure to achieve complete cures using current antibiotics. Specifically, several months after standard-care antibiotic therapy, a subset of patients can experience a series of persistent or relapsing symptoms, which are termed as chronic Lyme disease or post-treatment Lyme disease syndrome (PTLDS). While the persistence of the bacterial infection may or may not contribute to the etiology or pathogenesis of PTLDS, its treatment options remain unknown. Therefore, the development of vaccines and new drugs is highly warranted. We have identified a critical virulence determinant called BbHtrA that plays an indispensable role in host infectivity and persistence of spirochetes. Deletion of the protein renders the pathogen non- infectious in mammalian hosts. Based on this information, we propose a therapeutics strategy that relies on disruption of the protein function by small molecule drugs. To this end, we will identify small molecules that disrupt BbHtrA protease activity by employing an assay designed for high-throughput screening (HTS) of large compound libraries based on nanoDSF and fluorescence protease inhibition technology. Hits of this screen will be validated by a secondary cellular assay, revealing potent compounds that are permeable across the B. burgdorferi membrane. Compound screening will be performed in collaboration with National Center for Advancing Translational Sciences (NCATS) investigators, who provide expertise, infrastructure, and modern compound libraries, and have extensive experience with HTS and drug discovery. We will also attempt to develop structural studies on BbHtrA, which will be important for the development of new therapeutic agents targeting the protein. The validation of selected hit molecules in a whole animal model of Lyme disease will be examined in future larger grant applications. The project employs cutting-edge technology that targets interacting virulence determinants to combat Lyme disease, a novel strategy that may avoid PTLDS. Moreover, the same approach may serve as a paradigm for combating other tick-borne infections, including those caused by the recently-discovered, more virulent strains of Lyme disease pathogens.