Center for Microbial Pathogenesis, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, OH, 43205, USA.
Center for Microbial Pathogenesis, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, OH, 43205, USA; The Steve and Cindy Rasmussen Institute for Genomic Medicine, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, OH, 43205, USA.
EBioMedicine. 2024 Sep;107:105304. doi: 10.1016/j.ebiom.2024.105304. Epub 2024 Aug 24.
Bacterial biofilm communities are embedded in a protective extracellular matrix comprised of various components, with its' integrity largely owed to a 3-dimensional lattice of extracellular DNA (eDNA) interconnected by Holliday Junction (HJ)-like structures and stabilised by the ubiquitous eubacterial DNABII family of DNA-binding architectural proteins. We recently showed that the host innate immune effector High Mobility Group Box 1 (HMGB1) protein possesses extracellular anti-biofilm activity by destabilising these HJ-like structures, resulting in release of biofilm-resident bacteria into a vulnerable state. Herein, we showed that HMGB1's anti-biofilm activity was completely contained within a contiguous 97 amino acid region that retained DNA-binding activity, called 'mB Box-97'.
We engineered a synthetic version of this 97-mer and introduced a single amino acid change which lacked any post-translational modifications, and tested its activity independently and in combination with a humanised monoclonal antibody that disrupts biofilms by the distinct mechanism of DNABII protein sequestration.
mB Box-97 disrupted and prevented biofilms, including those formed by the ESKAPEE pathogens, and importantly reduced measurable proinflammatory activity normally associated with HMGB1 in a murine lung infection model.
Herein, we discuss the value of targeting the ubiquitous eDNA-dependent matrix of biofilms via mB Box-97 used singly or in a dual host-augmenting/pathogen-targeted cocktail to resolve bacterial biofilm infections.
This work was supported by NIH/NIDCD R01DC011818 to L.O.B. and S.D.G. and NIH/NIAID R01AI155501 to S.D.G.
细菌生物膜群落嵌入在由各种成分组成的保护性细胞外基质中，其完整性主要归因于细胞外 DNA（eDNA）的三维晶格，这些 DNA 通过类似于 Holliday 结（HJ）的结构相互连接，并由普遍存在的 eubacterial DNABII 家族的 DNA 结合结构蛋白稳定。我们最近表明，宿主先天免疫效应因子高迁移率族蛋白 B1（HMGB1）通过破坏这些类似于 HJ 的结构具有细胞外抗生物膜活性，导致生物膜内的细菌释放到脆弱状态。在此，我们表明 HMGB1 的抗生物膜活性完全包含在一个连续的 97 个氨基酸区域内，该区域保留了 DNA 结合活性，称为“mB Box-97”。
我们设计了这种 97 -mer 的合成版本，并引入了一个缺乏任何翻译后修饰的单一氨基酸变化，并独立测试了其活性，并与一种人源化单克隆抗体联合测试，该抗体通过 DNABII 蛋白隔离的独特机制破坏生物膜。
mB Box-97 破坏并阻止了生物膜的形成，包括 ESKAPEE 病原体形成的生物膜，并且重要的是，在小鼠肺部感染模型中降低了与 HMGB1 相关的可测量促炎活性。
在此，我们讨论了通过 mB Box-97 靶向生物膜中普遍存在的依赖于 eDNA 的基质的价值，无论是单独使用还是与单独使用或双重宿主增强/病原体靶向鸡尾酒一起使用，都可以解决细菌生物膜感染。
这项工作得到了 NIH/NIDCD R01DC011818 对 L.O.B. 和 S.D.G. 的资助，以及 NIH/NIAID R01AI155501 对 S.D.G. 的资助。
