ASSESSING SYNERGIES OF ANTIBACTERIAL PROTEINS AGAINST P AERUGINOSA BIOFILMS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 该项目将系统地测试这样一种假设，即通过与抗菌肽、人乳铁蛋白和/或Meveol的协同作用，作用于铜绿假单胞菌生物膜的基因工程人溶菌酶(HLYS)的抗菌性能得到增强。慢性下呼吸道感染导致细胞外、阴离子、生物聚合物的积累，如DNA、F-肌动蛋白、粘蛋白，在粘液型铜绿假单胞菌感染的情况下，胞外多糖海藻酸盐。这些生物聚合物在受感染的呼吸道中的高局部浓度被认为可以抑制阳离子抗菌肽和蛋白质，并且在人类溶菌酶(HLYS)功能方面尤其有问题。我们已经成功地重新设计了hLYS的静电潜力，在存在临床相关的抑制性生物聚合物的情况下，生产出具有增强杀菌活性的酶变体(2)。初步动物研究表明，与野生型蛋白或PBS对照相比，至少一种增强型人溶菌酶(EhLYS)可以减少肺部感染小鼠模型中铜绿假单胞菌的负担。将这项研究扩大到更大的临床影响，我们假设这种酶的抗假单胞菌活性将通过与互补的抗菌肽、抗菌蛋白人乳铁蛋白或Meveol共同给予而显著提高。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This project will systematically test the hypothesis that the antibacterial properties of genetically engineered human lysozyme (hLYS) acting on P. aeruginosa biofilms is enhanced by synergistic interactions with antimicrobial peptides, human lactoferrin, and/or Meveol. Chronic infection of the lower respiratory tract results in accumulation of extracellular, anionic, biopolymers such as DNA, F-actin, mucins, and in the case of mucoid P. aeruginosa infections, the exopolysaccharide alginate. The high local concentrations of these biopolymers in the infected airway are believed to inhibit cationic antimicrobial peptides and proteins, and are particularly problematic with respect to human lysozyme (hLYS) function. We have successfully re-engineered hLYS's electrostatic potential to produce enzyme variants with enhanced bactericidal activity in the presence of clinically relevant inhibitory biopolymers (2). Preliminary animal studies have indicated that at least one enhanced human lysozyme (EhLYS), when compared against the wild type protein or PBS controls, reduces P. aeruginosa burden in a mouse model of lung infection. Extending this research to have greater clinical impact, we hypothesize here that the anti-Pseudomonal activity of this enzyme will be substantially improved by co-administration with complementary antibacterial peptides, the antibacterial protein human lactoferrin, or Meveol.