Targeting Pneumococcal Virulence Factors in Otitis Media

针对中耳炎的肺炎球菌毒力因子

• 批准号: 8248737
• 负责人: HONGGAO YAN
• 金额: $ 18.35万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8248737
• 关键词: Affect; Affinity; Antibiotics; Antibodies; Applications Grants; Bacteremia; Binding; Binding Proteins; Binding Sites; Biochemical; Biomolecular Nuclear Magnetic Resonance; Biosensor; Catalytic DNA; Child; Choline; Communicable Diseases; Communities; Complement; Complement Factor H; Deposition; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Disease; Economic Burden; Effectiveness; Goals; Human; Human Factor H; Immunity; Infection; Laboratories; Laboratory Diagnosis; Lactoferrin; Length; Libraries; Life; Living Costs; Maps; Measures; Meningitis; Methods; Molecular; Molecular Biology; Molecular Target; NMR Spectroscopy; Nucleic Acid Binding; Otitis Media; Pathogenesis; Play; Pneumococcal Infections; Pneumonia; Production; Property; Protein Biochemistry; Proteins; Public Health; Research; Research Personnel; Resistance; Role; Scientist; Site; Specialist; Staging; Streptococcus pneumoniae; Testing; Therapeutic; Virulence Factors; Work; aptamer; bacterial genetics; base; cellular targeting; combat; ear infection; middle ear; novel; pathogen; pneumococcal surface protein A; public health relevance; tool

DESCRIPTION (provided by applicant): The long-term goal of the proposed research is to develop diagnostic tools for otitis media (OM) and other pneumococcal infections and research tools for studying pneumococcal pathogenesis. OM is a major public health problem in young children with an enormous economic burden in the US as well as in the world. S. pneumoniae not only is a major bacterial cause of OM but also causes additional life-threatening or invasive infections. Pneumococcal infection is one of the four killer infectious diseases and probably the world's single biggest killer of young children. Although S. pneumoniae is one of the most important human pathogens, the laboratory diagnosis of pneumococcal infections is still dependent on traditional microbiological methods and very few new methods are reliable enough for the laboratory diagnosis of pneumococcal infections. The traditional microbiological methods and tests not only are laborious but also have many limitations. There is an urgent need for new laboratory diagnostic tests for pneumococcal diseases, as the limitations of current diagnostic tests hinder the timely and accurate diagnosis of pneumococcal infections, which in turn negatively affects not only the treatment of the diseases but also the assessment of the effectiveness of control measures. Aptamers are small single-stranded nucleic acids that bind a molecular or cellular target with high affinity and their biochemical properties rival or are superior to those of antibodies in a variety of analytical, diagnostic, and potential therapeutic applications, particularly in ease of production, batch uniformity, shelf life, and cost. In this exploratory two-year grant application, we propose to develop aptamers against two major pneumococcal virulence factors, choline-binding protein A (CbpA, also known as PspC or SpsA) and pneumococcal surface protein A (PspA). Both virulence factors are multidomain multifunction proteins and play major roles in pneumococcal evasion of host immunity and pathogenesis. The molecular bases for their functions are the ability of CbpA to bind human complement factor H (FH) and the ectodomain of pIgR and the ability of PspA to bind apo- and holo-lactoferrin (LF) and to inhibit complement deposition. Specific Aim 1 is to develop aptamers against the virulence factors and Specific Aim 2 is to map where in the virulence factors the aptamers bind and develop aptamer beacons for the detection of the virulence factors. The proposed research will set up the stage for exploring the idea of using aptamers for the development of diagnostic tests for pneumococcal infections and provide research tools for visualizing pneumococcal pathogenesis. Based on the essential roles of pneumococcal virulence factors in pneumococcal pathogenesis and the ability of the developed aptamers to neutralize the two most important virulence factors, the proposed research will also set up the stage for testing the hypothesis of antivirulence strategy as a novel alternative/complementary strategy for combating pneumococcal resistance to classical antibiotics. PUBLIC HEALTH RELEVANCE:Otitis media, or middle ear infection, is a major public health problem in young children both in the US and in the world. Streptococcus pneumoniae not only is the major bacterial cause of otitis media but also causes additional life-threatening or invasive infections. The proposed research will set up the stage for exploring the idea of using aptamers for the development of diagnostic tests for pneumococcal infections and research tools for visualizing pneumococcal pathogenesis and for testing the hypothesis of antivirulence strategy as a novel potential strategy against otitis media and other pneumococcal infections.

描述（由申请人提供）：拟议的研究的长期目标是开发用于研究肺炎球菌发病机理的其他肺炎球菌感染和其他肺炎球菌感染和研究工具。 OM是在美国和世界上具有巨大经济负担的幼儿中的一个主要公共卫生问题。肺炎链球菌不仅是OM的主要原因，而且还会引起额外的威胁生命或侵入性感染。肺炎球菌感染是四种杀手传染病之一，也许是世界上最大的幼儿杀手。尽管肺炎链球菌是最重要的人类病原体之一，但肺炎球菌感染的实验室诊断仍然取决于传统的微生物学方法，并且很少有新方法足够可靠，足以实验室诊断肺炎球菌感染。传统的微生物方法和测试不仅艰苦，而且有很多局限性。迫切需要对肺炎球菌疾病进行新的实验室诊断测试，因为当前诊断测试的局限性阻碍了对肺炎球菌感染的及时，准确诊断，从而不仅对疾病的治疗产生了负面影响，而且对控制测量的有效性评估。适体是小的单链核酸，它们结合具有高亲和力的分子或细胞靶标，其生化特性竞争，或者在多种分析，诊断和潜在的治疗应用中，尤其是在生产，生产，批量统一性，固定性，固定性，固定性，固定性，固定性，和成本和成本中，都优于抗体的抗体。在此探索性的两年赠款应用中，我们建议针对两个主要的肺炎球菌毒力因子开发适体，胆碱结合蛋白A（CBPA，也称为PSPC或SPSA）和肺炎球菌表面蛋白A（PSPA）。两种毒力因子都是多域多功能蛋白，在宿主免疫和发病机理的肺炎球菌中起主要作用。其功能的分子碱基是CBPA结合人类补体因子H（FH）和PIGR的cBPA的能力，以及PSPA结合apo-和Holo-ractoferrin（LF）并抑制补体沉积的能力。具体目的1是开发针对毒力因子的适体，具体目的2是绘制适体在毒力因子中结合并开发适体信标，以检测毒力因子。拟议的研究将建立阶段，以探索使用适体开发肺炎球菌感染的诊断测试的想法，并提供可视化肺炎球菌发病机理的研究工具。基于肺炎球菌毒力因子在肺炎球菌发病机理中的基本作用，以及开发的适体中和中和中和两个最重要的毒力因子的能力，拟议的研究还将建立阶段，以测试将抗病毒性策略作为一种新型的替代/互补策略，作为一种新型的替代/互补策略，以使pneumocccal anticalccal anticalsics antical抗体抗体抗体。 公共卫生相关性：中耳炎或中耳感染是美国和世界上幼儿的主要公共卫生问题。肺炎链球菌不仅是中耳炎培养基的主要细菌原因，而且还会引起额外的威胁生命或侵袭性感染。拟议的研究将建立阶段，以探索使用适体开发肺炎球菌感染的诊断测试和研究工具，以可视化肺炎球菌发病机理，并测试将抗动力策略作为针对鼻炎媒体和其他肺炎球菌感染的新型潜在策略的假设。