权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Biomarker Screening of Essential Putative Glycoprotease Inhibitor

必需的假定糖蛋白酶抑制剂的生物标志物筛选

基本信息

批准号：
8259532
负责人：
Yinduo Ji
金额：
$ 37.37万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-15 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8259532
关键词：
Accounting Acetolactate Synthase Acids Acute suppurative arthritis due to bacteria Anti-Bacterial Agents Antibiotic Resistance Antibiotics Bacteria Biochemical Biological Assay Biological Markers Biological Process Cell Wall Cells Chemicals Communities Community Hospitals DNA Development Down-Regulation Drug Delivery Systems Drug Industry Endocarditis Enzymes Genes Goals Growth Haemophilus influenzae Health care facility Homologous Gene Hydro-Lyases In Vitro Infection Intermediate resistance Lead Life Luciferases Medical Metabolic Pathway Molecular Target Monitor Multi-Drug Resistance Mycoplasma genitalium Nosocomial Infections Outcome Pathogenicity Patients Pharmaceutical Preparations Phenotype Plasmids Pneumonia Preventive Production Proteins Proteomics Public Health RNA Interference Reporter Research Resistance Rice Screening procedure Set protein Skin Specificity Staphylococcus aureus Streptococcus pneumoniae Surrogate Markers System Systemic infection Technology Testing Therapeutic Agents Therapeutic Intervention Threonine Dehydratase Toxic Shock Syndrome Toxic effect Vaccines Validation Vancomycin Vancomycin-resistant S. aureus antimicrobial base cellular targeting cost fighting high throughput screening inhibitor/antagonist methicillin resistant Staphylococcus aureus novel novel strategies novel therapeutics pathogen promoter public health relevance resistant strain therapeutic target tool

项目摘要

DESCRIPTION (provided by applicant): Given the well-known pathogenicity of Staphylococcus aureus, the emergence of methicillin-resistant S. aureus (MRSA) and vancomycin-resistant S. aureus (VRSA) is causing increased public health concerns. More than 60% of S. aureus-associated hospital acquired infections are caused by MRSA, which has increasingly spread beyond healthcare facilities and has emerged as a community pathogen. The annual cost to treat MRSA in hospitalized patients in the U.S. is estimated to be between $3.2 billion to 4.2 billion. The threat of VRSA has been the topic of intensive research since there is a limited drug of choice for MRSA infections. Therefore, there is an urgent need to develop more efficient preventive and/or therapeutic agents to fight this pathogen. We have demonstrated that a staphylococcal putative glycoprotease (Gcp) is a unique antibacterial target since it is cell-wall associated and secreted, essential for survival for both Gram-positive and Gram-negative pathogens. However, the biological function of the putative glycoprotease is still unclear. Without a biochemical readout phenotype, it is difficult for us to develop a high-throughput screen of inhibitors against this novel target. Combined microarray and proteomic approaches with a regulated RNA interference technology, we found that the down-regulation of gcp expression significantly increases the productions of acetolactate synthase, dihydroxy-acid dehydratase and threonine dehydratase, which are located in the same metabolic pathway in S. aureus. Therefore, we hypothesize that these enzymes are the surrogate markers of Gcp and monitoring Gcp's biomarkers will provide a powerful approach to high-throughput screen inhibitors against Gcp. The objective of this proposal is to develop a whole cell-based high throughput assay using these elevated proteins as Gcp's surrogate biomarkers. In specific aim 1, we will construct bioluminescent reporter systems using Gcp's surrogate markers in S. aureus. In specific aim 2, we will perform high-throughput screening of novel chemical leads against this essential putative glycoprotease using the bioluminescent reporter systems. In specific aim 3, we will validate lead compounds to determine their mode of action, specificity, and selectivity. Successful completion of these studies will identify potential inhibitors against this novel, unique antibacterial target, Gcp, as well as provide a novel strategy for high-throughput screening of compounds against other functional unknown targets. Therefore, the proposed studies will provide a novel strategy for high-throughput screening of compounds against novel drug target that will be particularly important to therapeutic interventions for infections and medical practice to fight the emerging antibiotic-resistant S. aureus. PUBLIC HEALTH RELEVANCE: S. aureus is an important community and hospital acquired pathogen causing a wide variety of infections, ranging from superficial skin to life-threatening systemic infections, including pneumonia, endocarditis, septic arthritis and toxic shock syndrome. The recent emergence of multiple antibiotic- resistant strains of S. aureus, especially methicillin-resistant S. aureus and vancomycin-intermediate resistant S. aureus, is causing great concern in the public health community. There is an urgent need for new classes of antibiotics and potent vaccines to fight infections caused by S. aureus. The outcome of our proposed project will identify potential lead compounds against a unique, secreted antibacterial target, as well as provide a novel strategy for high-throughput screening of inhibitors against functional unknown targets.

描述（由申请人提供）：鉴于金黄色葡萄球菌众所周知的致病性，耐甲氧西林金黄色葡萄球菌（MRSA）和耐万古霉素金黄色葡萄球菌（VRSA）的出现正在引起越来越多的公众健康担忧。超过 60% 的金黄色葡萄球菌相关医院获得性感染是由 MRSA 引起的，这种细菌越来越多地传播到医疗机构之外，并已成为一种社区病原体。在美国，每年治疗住院患者 MRSA 的费用估计在 32 亿至 42 亿美元之间。由于治疗 MRSA 感染的药物有限，VRSA 的威胁一直是深入研究的主题。因此，迫切需要开发更有效的预防和/或治疗剂来对抗这种病原体。我们已经证明，葡萄球菌假定的糖蛋白酶（Gcp）是一种独特的抗菌靶点，因为它与细胞壁相关并分泌，对于革兰氏阳性和革兰氏阴性病原体的生存至关重要。然而，假定的糖蛋白酶的生物学功能仍不清楚。如果没有生化读数表型，我们就很难针对这一新靶标开发高通量的抑制剂筛选。将微阵列和蛋白质组学方法与受调控的RNA干扰技术相结合，我们发现gcp表达的下调显着增加了乙酰乳酸合酶、二羟酸脱水酶和苏氨酸脱水酶的产生，这些酶位于金黄色葡萄球菌中的同一代谢途径中。因此，我们假设这些酶是 Gcp 的替代标记，监测 Gcp 的生物标记将为高通量筛选 Gcp 抑制剂提供强大的方法。该提案的目的是开发一种基于全细胞的高通量测定，使用这些升高的蛋白质作为 Gcp 的替代生物标志物。在具体目标 1 中，我们将使用 Gcp 的金黄色葡萄球菌替代标记构建生物发光报告系统。在具体目标 2 中，我们将使用生物发光报告系统对这种重要的假定糖蛋白酶的新型化学先导物进行高通量筛选。在具体目标 3 中，我们将验证先导化合物以确定其作用方式、特异性和选择性。这些研究的成功完成将确定针对这种新颖、独特的抗菌靶标 Gcp 的潜在抑制剂，并为高通量筛选针对其他功能未知靶标的化合物提供新策略。因此，拟议的研究将为针对新药物靶标的化合物的高通量筛选提供新策略，这对于感染的治疗干预和对抗新出现的抗生素耐药性金黄色葡萄球菌的医疗实践尤为重要。公共卫生相关性：金黄色葡萄球菌是一种重要的社区和医院获得性病原体，可引起多种感染，从浅表皮肤到危及生命的全身感染，包括肺炎、心内膜炎、化脓性关节炎和中毒性休克综合征。近年来出现的多重耐药金黄色葡萄球菌菌株，特别是耐甲氧西林金黄色葡萄球菌和耐万古霉素金黄色葡萄球菌，引起了公共卫生界的高度关注。迫切需要新型抗生素和强效疫苗来对抗金黄色葡萄球菌引起的感染。我们提出的项目的结果将确定针对独特的、分泌的抗菌靶标的潜在先导化合物，并为高通量筛选针对功能性未知靶标的抑制剂提供新策略。