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Co-opting Endogenous Pathogen Autolysins as Next Generation Antibiotics

选择内源性病原体自溶素作为下一代抗生素

基本信息

批准号：
10053699
负责人：
Karl E Griswold
金额：
$ 54.77万
依托单位：
DARTMOUTH COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-11-18 至 2022-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10053699
关键词：
Acinetobacter Address Anti-Bacterial Agents Antibiotic Resistance Antibiotics Autolysin Bacteria Bacterial Drug Resistance Bacterial Infections Bacterial Physiology Bacteriophages Bioinformatics Biological Assay Biological Response Modifier Therapy British Cell Wall Centers for Disease Control and Prevention (U.S.)Cessation of life Clinical Cytolysis Development Directed Molecular Evolution Drug resistance ESKAPE pathogens Engineering Enterobacteriaceae Enterococcus faecium Enzymes Exhibits Fluorescence-Activated Cell Sorting Future Gel Genetic Engineering Genome Genomic Library Genus staphylococcus Goals Government Harvest Health Human Hydrolase Immune system In Vitro Klebsiella pneumoniae Lead Libraries Life LytA enzyme Lytic Maintenance Metagenomics Methods Molecular Muramidase Nature Organism Pathogenicity Pathway interactions Peptidoglycan Performance Pharmaceutical Preparations Phenotype Predisposition Process Proteome Pseudomonas aeruginosa Recombinants Refractory Reporting Resistance Resistance development Specificity Staphylococcus aureus Testing Therapeutic Trust Ursidae Family Work antimicrobial drug bacterial resistance bacteriocin burden of illness cellular targeting chemotherapy clinically relevant combinatorial cost drug resistant bacteria endolysin enzyme activity enzyme therapy experience experimental study high throughput screening improved in vivo innovation lead candidate lead optimization methicillin resistant Staphylococcus aureus microbial next generation novel novel therapeutics pathogen pathogenic bacteria resistant strain screening side effect small molecule

项目摘要

Summary: Antibiotic resistance represents one of the greatest threats to human health. In particular, the six so-called ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumanii, Pseudomonas aeruginosa, and enterobacteriaceae) represent highly drug-resistant bacteria that exert a tremendous global burden of disease. The potential scope of this crisis was highlighted in a recent report commissioned by the Wellcome Trust and British Government; the authors projected that, by 2050, drug-resistant bacterial infections could cost the global economy a cumulative $100 trillion and kill 10 million people annually. To address this issue, there is a critical need for innovative antibacterial treatments. One compelling therapeutic strategy leverages recombinant enzymes that degrade cell wall peptidoglycan, thereby causing bacterial lysis and death. Currently, all such lytic enzyme therapies are trans-acting in nature, i.e., they are derived from bacteriophage or the immune systems of eukaryotic organisms. This proposal seeks to establish an entirely new paradigm for developing bacteriolytic enzyme drugs. We hypothesize that a pathogen's own endogenous cell wall hydrolases (i.e., “autolysins”) can be co-opted to yield potent antimicrobial agents that are refractory to new resistance phenotypes. To test this hypothesis, we will pursue initial studies with the high impact pathogen methicillin resistant S. aureus (MRSA), although the strategy should be broadly applicable to any bacterial pathogen. Here, complementary computational and experimental approaches will be utilized to identify, isolate, and engineer potent autolysins derived from staphylococcal proteomes. In aim 1, the sequenced genome of S. aureus and related bacteria will be searched for autolysins using bioinformatics. Candidate enzymes will be cloned, evaluated, and their activities will be improved via computationally guided fusion to high performance cell wall targeting domains. In aim 2, a complementary high throughput screening strategy will be taken to identify autolysins from genomic libraries of pathogenic staphylococci. The activities of candidate enzymes will be improved via combinatorial chimeragenesis with high performance cell wall targeting domains, followed by high throughput functional screening of the resultant chimeric libraries. In aim 3, lead autolysin candidates will be further engineered for potent anti-staphylococcal activity using a directed evolution strategy. The most promising lead candidates from these studies will be rigorously evaluated using a panel of clinically relevant in vitro and in vivo assays. Ultimately, this project could yield both novel anti-staphylococcal agents and an entirely new paradigm for development of antibacterial biotherapies.

概括：抗生素耐药性是对人类健康的最大威胁之一。特别是，所谓的六 ESKAPE 病原体（屎肠球菌、金黄色葡萄球菌、肺炎克雷伯菌、不动杆菌鲍曼不动杆菌、铜绿假单胞菌和肠杆菌科）代表高度耐药的细菌造成巨大的全球疾病负担。最近的一份报告强调了这场危机的潜在范围受威康信托基金会和英国政府委托编写的报告；作者预测，到 2050 年，耐药细菌感染可能给全球经济造成累计 100 万亿美元的损失，并导致 1000 万人死亡人每年。为了解决这个问题，迫切需要创新的抗菌治疗。一令人信服的治疗策略利用重组酶降解细胞壁肽聚糖，从而导致细菌裂解和死亡。目前，所有此类裂解酶疗法本质上都是反式作用的，即它们源自噬菌体或真核生物的免疫系统。该提案旨在建立了开发溶菌酶药物的全新范例。我们假设一个病原体自身的内源性细胞壁水解酶（即“自溶素”）可以被选择产生有效的对新的耐药表型难以抵抗的抗菌药物。为了检验这个假设，我们将追求针对高影响病原体耐甲氧西林金黄色葡萄球菌 (MRSA) 的初步研究，尽管该策略应广泛适用于任何细菌病原体。在这里，互补的计算和实验将利用方法来识别、分离和设计源自葡萄球菌的有效自溶素蛋白质组。在目标 1 中，将搜索金黄色葡萄球菌和相关细菌的测序基因组中的自溶素使用生物信息学。候选酶将被克隆、评估，并通过以下方式提高其活性计算引导融合到高性能细胞壁靶向域。在目标 2 中，互补性高将采取通量筛选策略从致病性基因组文库中鉴定自溶素葡萄球菌。候选酶的活性将通过与以下物质的组合嵌合得到改善：高性能细胞壁靶向结构域，然后对所得结果进行高通量功能筛选嵌合文库。在目标 3 中，将进一步设计先导自溶素候选物，以实现有效的抗葡萄球菌作用使用定向进化策略的活动。这些研究中最有希望的主要候选者将是使用一组临床相关的体外和体内测定进行严格评估。最终，这个项目可以产生新型抗葡萄球菌药物和抗菌剂开发的全新范例生物疗法。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Bioinformatics-driven discovery of novel Clostridioides difficile lysins and experimental comparison with highly active benchmarks.

DOI：
10.1002/bit.27759
发表时间：
2021-07
期刊：
BIOTECHNOLOGY AND BIOENGINEERING
影响因子：
3.8
作者：
Furlon, Jacob M.;Mitchell, Spencer J.;Bailey-Kellogg, Chris;Griswold, Karl E.
通讯作者：
Griswold, Karl E.

Nonclassical antagonism between human lysozyme and AMPs against Pseudomonas aeruginosa.