Optimizing the immune response by targeting the Sts enzymes

项目摘要 Sts磷酸酶负调节哺乳动物免疫细胞内的信号传导途径。 系统缺乏Sts表达的小鼠（Sts-/-）对临床相关真菌感染具有深刻的抗性 和细菌病原体，包括白色念珠菌、土拉热弗朗西斯菌和金黄色葡萄球菌。 耐药性与快速清除病原体和减少炎症有关。我们的初步数据 表明缺乏Sts表达的吞噬细胞具有增强的杀微生物功能。 我们已经证明Sts磷酸酶结构域在结构和机制上与其他磷酸酶结构域不同。 蛋白磷酸酶的种类。我们还表明，Sts活性位点位于一个独特的结合， 口袋，酶活性可以被药物样化合物竞争性抑制。我们假设 药物介导的Sts-1抑制将重现Sts-/-表型，并导致有益的临床应用。 结果。根据这一假设，本提案的目的是确定和描述小规模 Sts-1的分子抑制剂，其可增强白细胞抗微生物应答并显示功效 在整个动物感染模型中。为了实现这一目标，我们将进行高通量筛选 (HTS)530 K化合物Scripps药物发现库。活性化合物将广泛用于 使用我们建立的测定和体内模型进行表征和彻底验证。 我们将通过完成以下具体目标来实现我们的目标： 具体目标1：进行530 K化合物HTS以鉴定Sts-1磷酸酶活性的抑制剂。 具体目标2：鉴定和表征命中化合物。 具体目标3：确定Sts抑制性化合物对宿主对微生物的应答的影响 感染 成功完成拟议的研究将产生一套经验证的小分子Sts抑制剂 磷酸酶活性，这将作为功能的化学探针，并作为开发的线索 新的治疗药物。从长远来看，这项工作预计将为 制定新的临床方案，将显著降低归因于 系统性病原体感染，通过增强宿主免疫反应。

PROJECT SUMMARY The Sts phosphatases negatively regulate signaling pathways within cells of the mammalian immune system. Mice lacking Sts expression (Sts-/-) are profoundly resistant to infection by clinically relevant fungal and bacterial pathogens, including Candida albicans, Francisella tularensis, and Staphylococcus aureus. Resistance is associated with rapid pathogen clearance and reduced inflammation. Our preliminary data demonstrates that phagocytes lacking Sts expression have enhanced microbicidal functions. We have shown that the Sts phosphatase domain is structurally and mechanistically distinct from other classes of protein phosphatases. We also demonstrate that the Sts active site sits in a distinct binding pocket, and enzyme activity can be competitively inhibited with drug-like compounds. We hypothesize that drug-mediated inhibition of Sts-1 will recapitulate the Sts-/- phenotype and lead to beneficial clinical outcomes. Based on this hypothesis, the objective of this proposal is to identify and characterize small molecule inhibitors of Sts-1 that can enhance leukocyte anti-microbial responses and demonstrate efficacy in whole animal infection models. To achieve this objective, we will conduct a high-throughput screen (HTS) of the 530K compound Scripps Drug Discovery Library. Active compounds will be extensively characterized and thoroughly validated, using our established assays and in vivo models. We will accomplish our objective by completing the following Specific Aims: Specific Aim 1: Conduct a 530K compound HTS to identify inhibitors of Sts-1 phosphatase activity. Specific Aim 2: Validate and characterize hit compounds. Specific Aim 3: Determine the effects of Sts inhibitory compounds on host responses to microbial infection. Successful completion of the proposed studies will yield a set of validated small molecule inhibitors of Sts phosphatase activity that will serve both as chemical probes of function and as leads for the development of novel therapeutic agents. In the long term, this work is expected to provide a foundation for the development of new clinical protocols that will significantly reduce the morbidity and mortality attributed to systemic pathogen infections, by enhancing host immune responses.