Recombinant Fc fusions for treatment of uropathogenic E. coli

用于治疗尿路致病性大肠杆菌的重组 Fc 融合体

基本信息

批准号：
10021217
负责人：
EVGENI Veniaminovic SOKURENKO
金额：
$ 48.69万
依托单位：
PLANET BIOTECHNOLOGY, INC.
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10021217
关键词：
Adhesions Affect Animal Model Antibiotic Resistance Antibiotics Antibodies Bacteria Bacterial Adhesins Bacterial Adhesion Bacterial Attachment Site Bacterial Infections Binding Binding Proteins Biological Assay Budgets C3bi Cell Surface Receptors Cell-Matrix Junction Cells Classical Complement Pathway Clinical Trials Complement Complement 1q Complement 3b Complement Activation Complement Membrane Attack Complex Complement-Dependent Cytotoxicity Dipeptidyl Peptidases Enterobacter Enterobacteriaceae Escherichia coli Escherichia coli Infections Frequencies Future Galactose Goals Human Hybrids IgG1 IgG3 Immune Immunoglobulin G In Vitro Klebsiella Klebsiella Infections Klebsiella pneumoniae Macrophage-1 Antigen Mannose Mediating Membrane Membrane Glycoproteins Modification Molecular Multi-Drug Resistance N-terminal Phagocytes Phagocytosis Phase Plants Point Mutation Polysaccharides Protein Fragment Proteins Public Health Recombinant Proteins Recombinants Research Project Grants Resistance Serum Small Business Innovation Research Grant Surface Testing Urinary tract infection Uropathogenic E. coli Urothelial Cell Urothelium Variant Virulence Factors bacterial resistance community-acquired UTI cost glycosylation hybrid protein improved in vitro activity monomer neutrophil novel pandemic disease pathogen pathogenic bacteria resistant strain type 1 fimbriae

项目摘要

Urinary tract infections (UTIs) are among the most common bacterial infections, affecting 150 million people worldwide each year. In the USA, community-acquired UTIs account for about 11 million cases each year that cost the U.S. public health budget $5 billion annually. Uropathogenic Escherichia coli (UPEC) accounts for up to 80% of UTIs. While UTIs are currently treated with antibiotics, the frequency of multi-drug resistance is increasing, portending a future of untreatable UTIs. A promising alternative to antibiotics is to directly target bacterial virulence factors that are critical for bacterial adhesion to and invasion of uroepithelial cells. These virulence factors include the bacterial adhesin FimH, expressed by the majority of uropathogenic E. coli strains. FimH binds to a specific glycoform, oligomannose-3, carried by an abundant membrane glycoprotein on uroepithelial cells. We have produced a recombinant protein targeting FimH. It is a molecular fusion of human protein, dipeptidyl peptidase 4 (DPP4) with IgG1 Fc (DPP4-Fc), carries eight mannose-containing N-glycans and binds in a mannose-specific manner to FimH. We have shown that that DPP4-Fc promotes killing of uropathogenic E. coli by complement. The purpose of this research project is to demonstrate the ability of DPP4-Fc to promote activation of complement on the surface of antibiotic-resistant uropathogenic E. coli thereby leading to increased bacterial killing by complement and polymorphonuclear cells (PMNs). We will produce three new variants of DPP4-Fc carrying different numbers of N-glycans per monomer and different glycoforms. We will modify the Fc of DPP4-Fc to improve C1q binding and thus increase activation of the classical complement pathway. Modifications will include point mutations and replacement of IgG1 Fc with IgG3 Fc. We will also produce a negative control DPP4-Fc in which the Fc contains two point mutations that abrogate C1q binding and thus has no opsonizing activity. We will test all these new DPP4-Fc variants for their ability to: i) bind to uropathogenic E.coli, Enterobacter and Klebsiella pneumonia strains, ii) inhibit bacterial attachment to urothelial cells, iii) promote serum mediated killing of bacteria and iv) promote opsonophagocytosis of bacteria. We expect that DPP4-Fc can be developed into a novel treatment for uropathogenic E. coli, Enterobacter and Klebsiella infections. Future research projects will test the activity of these molecules in animal models and later in human clinical trials.

尿路感染（UTI）是最常见的细菌感染之一，影响1.5亿每年在全球范围内的人们。在美国，社区获得的UTI每年约有1100万个案例这损失了美国公共卫生预算每年50亿美元。肝病大肠杆菌（UPEC）占多达80％的UTI。虽然目前用抗生素处理UTI，但多药耐药性的频率为增加，适用于无法治疗的UTI的未来。有希望的抗生素替代方法是直接靶向细菌毒力因子对细菌粘附至关重要的细菌粘附和浸润性细胞至关重要。这些毒力因子包括大多数肝癌大肠杆菌菌株表达的细菌粘附素FIMH。 FIMH与特定的糖型，寡素糖-3结合，该糖蛋白在紫色细胞。我们产生了靶向FIMH的重组蛋白。它是人类蛋白的分子融合， IgG1 FC（DPP4-FC）的二肽基肽酶4（DPP4）携带八个含甘露糖的N-聚糖并结合以特定于甘露糖的方式进行fimh。我们已经表明，DPP4-FC促进了尿路病的杀死。大肠杆菌补充。该研究项目的目的是证明DPP4-FC促进的能力在抗生素耐药性肝癌大肠杆菌表面的补体激活，从而导致增加通过补体和多形核细胞（PMN）杀死细菌。我们将生成三种新的DPP4-FC变体，每个单体携带不同数量的N-聚糖，并且不同的糖型。我们将修改DPP4-FC的FC以改善C1Q结合，从而增加激活的激活经典的补体途径。修改将包括点突变和用IgG1 FC替换 IgG3 FC。我们还将产生一个阴性对照DPP4-FC，其中FC包含两个点突变，废除C1Q结合，因此没有打击活性。我们将测试所有这些新的DPP4-FC变体能力：i）与尿道病大肠杆菌结合，肠杆菌和克雷伯菌肺炎菌株，ii）抑制细菌附着在尿路上皮细胞上，iii）促进血清介导的细菌杀死和iv）促进肠胃病细菌。我们预计DPP4-FC可以发展为肝癌大肠杆菌肠杆菌的新型治疗方法和克雷伯菌感染。未来的研究项目将测试这些分子在动物模型中的活性，并后来在人类的临床试验中。