Endolysins as tools to eradicate pneumococcal biofilms and development of protective immunity

内溶素作为根除肺炎球菌生物膜和发展保护性免疫的工具

基本信息

批准号：
10587285
负责人：
Norberto Gonzalez Juarbe
金额：
$ 90.1万
依托单位：
J. CRAIG VENTER INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-12-09 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10587285
关键词：
Address Animal Model Animals Antibiotic Resistance Antibiotics Antibodies Antibody Formation Antibody Response Antibody-mediated protection Antigens Antimicrobial Resistance Apoptosis Automobile Driving Bacteremia Bacteria Bacterial Proteins Bacteriophages Benchmarking Biochemical Biochemistry Biological Biological Assay Biophysics Cell Death Cell Death Induction Cells Cessation of life Child Coculture Techniques Country Developing Countries Development Disadvantaged Disease Effectiveness Elderly Enzymes Epithelium Extracellular Matrix Fatality rate Generations Goals Gram-Positive Bacteria Health Health Care Costs Human Immunity Immunize Immunologic Techniques Immunologics In Vitro Individual Inflammation Lead Lung Lytic Phase Macrophage Mass Spectrum Analysis Microbial Biofilms Modeling Molecular Mus N-Acetylmuramoyl-L-alanine Amidase Nasopharynx Necrosis Nose Oral mucous membrane structure Organism Otitis Media Peptidoglycan Pharyngeal structure Phosphotransferases Pneumococcal Colonization Pneumococcal Infections Pneumonia Polymers Polysaccharides Population Predisposition Property Proteins Proteomics Proxy Research Resistance Role Safety Serotyping Sinusitis Specificity Streptococcus pneumoniae Streptococcus pneumoniae plY protein Technology Testing Therapeutic Tissues Toxic effect Toxin Transgenic Mice Vaccines World Health Organization antimicrobial aspirate cellular development community acquired pneumonia effectiveness evaluation effectiveness testing endolysin extracellular immunogenic in vivo member mucosal vaccine next generation novel opportunistic pathogen oral commensal point of care prevent resistant strain response single-cell RNA sequencing sugar tool transcriptomics

项目摘要

Project Summary Streptococcus pneumoniae (Spn, the pneumococcus) are Gram-positive bacteria and the leading cause of community-acquired pneumonia worldwide. The World Health Organization estimates >1.6 million deaths are the result of Spn infection each year, with children and the elderly being the most susceptible populations. Major problems of pneumococcal disease include the acquisition of antimicrobial resistance and the global spread of resistant clones. In addition, these problems are magnified by the major disadvantages of the current capsular polysaccharide-based vaccines, such as serotype specificity and the resulting incomplete coverage. An emerging way to address the growing antimicrobial resistance problem is the use of bacteriophage endolysins. These enzymes are capable of degrading the bacterial peptidoglycan, killing and dispersing biofilm bacteria and its matrix. In preliminary studies, we have developed a chimeric derivative of the well characterized Cpl-1 endolysin that displays >100-fold increase in antimicrobial activity, termed ClyX-1. We also show the ability of this new endolysin to lyse planktonic Spn, and importantly we also observed that ClyX-1 was able to kill biofilm Spn, as well as disperse the biofilm matrix. In addition, we have shown that upon Spn nasopharyngeal colonization, activation of programmed necrosis, i.e. necroptosis, leads to development of antigen-specific antibodies. Of note, treatment of colonized mice with ClyX-1 further promoted necroptosis activation, suggesting endolysin treatment may enhance development of protective immunity against Spn. Herein, we aim to address three overall hypotheses: a) that endolysins are efficient pneumococcal anti-biofilm agents, b) that endolysins can be an effective way to prevent Spn colonization in a serotype-independent manner, and c) that intranasal treatment with endolysins promotes protective immunity, to prevent re-colonization and severe disease. We will use a combination of in vitro and in vivo studies with static and dynamic biofilms, mice, biochemistry (characterize and benchmark pneumococcal endolysins), transgenic mice (to define the role of programmed cell death in protective immunity), molecular and immunological techniques and next generation technologies (proteomics, single cell transcriptomics) to establish better understanding of the effects of endolysin treatments against pneumococcal disease in vivo and test their effectiveness in development of long-term serotype-independent protective immunity.

项目摘要肺炎链球菌（SPN，肺炎球菌）是革兰氏阳性细菌，是导致的主要原因全球社区获得的肺炎。世界卫生组织估计死亡> 160万 SPN感染的结果是，儿童和老年人是最敏感的人群。主要的肺炎球菌疾病的问题包括获得抗菌素耐药性和全球蔓延抗性克隆。另外，这些问题被当前囊的主要缺点放大基于多糖的疫苗，例如血清型特异性和结果不完整的覆盖范围。一个解决日益增长的抗菌耐药性问题的新兴方法是使用噬菌体内olysins。这些酶能够降解细菌肽聚糖，杀死和分散生物膜细菌，并它的矩阵。在初步研究中，我们已经开发了一个表征良好的CPL-1的嵌合导数内olysin在抗菌活性中显示出> 100倍，称为CLYX-1。我们还展示了这种新的内olysin裂解浮游SPN，重要的是，我们还观察到Clyx-1能够杀死生物膜 SPN以及分散生物膜基质。此外，我们已经表明，在SPN鼻咽上定殖，激活程序坏死，即坏死，导致抗原特异性的发展抗体。值得注意的是，用Clyx-1治疗定植的小鼠进一步促进了坏死性激活，表明内olysin治疗可以增强对SPN的保护性免疫的发展。在此，我们的目标是解决三个总体假设：a）内叶蛋白是有效的肺炎球菌抗双膜剂，b）内叶蛋白可以是一种以血清型独立的方式预防SPN定植的有效方法，c）鼻内用内叶蛋白治疗可促进保护性免疫，以防止再殖民化和严重疾病。我们将使用静态和动态生物膜，小鼠，生物化学的体外研究和体内研究（表征）和基准肺炎球菌内olysins），转基因小鼠（定义了程序性细胞死亡在保护性免疫），分子和免疫学技术以及下一代技术（蛋白质组学，单细胞转录组学），以更好地了解内olysin治疗的影响体内肺炎球菌疾病并测试其在长期型独立的发展中的有效性保护性免疫。