Novel gene-silencing therapeutics for multidrug-resistant gram-negative pathogens

项目概要/摘要 对新型抗菌剂的需求日益迫切。多药耐药病原体的比率继续上升， 增加，导致世界各地的发病率和死亡率大幅上升。此外，目前的管道 用于新的抗菌剂仍然非常狭窄。美国传染病协会在他们的研究中发现， “坏虫子，没有药物”运动，一组病原体，已成为越来越多的耐药电流 抗生素该组包括革兰氏阴性病原体鲍氏不动杆菌和大肠杆菌。 抗生素发现和设计的新范式最近已被证明对许多 细菌这种新方法基于一种名为肽-磷酰二胺mopholino的平台技术 低聚物（PPMOs）。PPMO是合成的DNA模拟物，其以序列特异性反义寡核苷酸结合RNA。 方式并抑制细菌必需基因的表达。PPMO已经成功地用于杀死 多种细菌病原体，包括革兰氏阴性菌大肠杆菌、沙门氏菌 鼠伤寒沙门氏菌、洋葱伯克霍尔德氏菌复合菌和鲍氏不动杆菌。PPMOS在培养物中是杀菌的， 并减少菌血症和提高感染动物模型的存活率。PPMOs的效力比 许多传统抗生素如氨苄青霉素。该项目的目标是开发PPMO用于治疗 用于对抗多重耐药病原体大肠杆菌和鲍曼不动杆菌。具体 目的是设计，生产和筛选针对多药耐药细胞中各种基因靶点的PPMOS， 病原体E. coli和A.鲍曼不动杆菌。实验方法是靶向基因的途径， 已知或怀疑是生物体生长所必需的，包括生物合成的基因。 脂多糖、肽聚糖和脂肪酸。另一种方法是使用PPMO作为替代品， 治疗和靶向特定的抗生素耐药机制，以恢复对目前 使用抗生素。这种技术提供了一种方法上的优势，因为许多PMOs可以快速地 合成并同时针对多个目标进行测试。这就有可能瞄准 单一生物体中的多个基因或针对多种病原体的PPMO鸡尾酒的发展。 本项目将确定E. coli和A.鲍曼不动杆菌可以被转移到前 临床和临床研究。

Project Summary/Abstract The need for new antimicrobials is increasingly urgent. The rate of multidrug resistant pathogens continues to increase leading to significant morbidity and mortality throughout the world. Furthermore, the current pipeline for new antimicrobials remains very narrow. The Infectious Diseases Society of America has identified in their "Bad Bugs, No Drugs" campaign, a group of pathogens that have become increasingly resistant to current antibiotics. This group includes the Gram-negative pathogens Acinetobacter baumannii and Escherichia coli. A new paradigm in antibiotic discovery and design has recently been shown effective against numerous bacteria. This new approach is based on a platform technology called peptide-phosphorodiamidate mopholino oligomers (PPMOs). PPMOs are synthetic DNA mimics that bind to RNA in a sequence-specific, antisense manner and inhibit expression of essential bacterial genes. PPMOs have already been used successfully to kill a variety of bacterial pathogens including the Gram-negative bacteria Escherichia coli, Salmonella typhimurium, Burkholderia cepacia complex and Acinetobacter baumannii. PPMOS are bactericidal in culture, and reduce bacteremia and improve survival in animal models of infection. PPMOs are more potent than many traditional antibiotics such as ampicillin. The goal of this project is to develop PPMOs for therapeutic use against the multidrug resistant pathogens Escherichia coli and Acinetobacter baumannii. The specific aims are to design, produce and screen PPMOS against various gene targets in the multidrug-resistant pathogens E. coli and A. baumannii. The experimental approach is to target genes in pathways that are known or suspected to be essential for the growth of the organism, including genes for biosynthesis of lipopolysaccharide, peptidoglycan, and fatty acids. Another approach will be to use PPMOs as adjunctive therapies and target specific antibiotic resistance mechanisms in order to restore susceptibility to currently used antibiotics. This technology provides a methodological advantage because many PPMOs can be rapidly synthesized and simultaneously tested against numerous targets. This allows for the possibility of targeting multiple genes in a single organism or the development of cocktails of PPMOs that target multiple pathogens. This project will identify lead target PPMOs in E. coli and A. baumannii that can be moved forward to pre- clinical and clinical studies.