Institute of Food, Nutrition and Health, ETH Zurich, Zurich, Switzerland.
Department of Infectious Diseases and Hospital Epidemiology, University Hospital Zurich, University of Zurich, Zurich, Switzerland.
mBio. 2020 Apr 14;11(2):e00209-20. doi: 10.1128/mBio.00209-20.
is a major concern in human health care, mostly due to the increasing prevalence of antibiotic resistance. Intracellular localization of plays a key role in recurrent infections by protecting the pathogens from antibiotics and immune responses. Peptidoglycan hydrolases (PGHs) are highly specific bactericidal enzymes active against both drug-sensitive and -resistant bacteria. However, PGHs able to effectively target intracellular are not yet available. To overcome this limitation, we first screened 322 recombineered PGHs for staphylolytic activity under conditions found inside eukaryotic intracellular compartments. The most active constructs were modified by fusion to different cell-penetrating peptides (CPPs), resulting in increased uptake and enhanced intracellular killing (reduction by up to 4.5 log units) of various strains (including methicillin-resistant [MRSA]) in different tissue culture infection models. The combined application of synergistic PGH-CPP constructs further enhanced their intracellular efficacy. Finally, synergistically active PGH-CPP cocktails reduced the total by more than 2.2 log units in a murine abscess model after peripheral injection. Significantly more intracellular bacteria were killed by the PGH-CPPs than by the PGHs alone. Collectively, our findings show that CPP-fused PGHs are effective novel protein therapeutics against both intracellular and drug-resistant The increasing prevalence of antibiotic-resistant bacteria is one of the most urgent problems of our time. is an important human pathogen that has acquired several mechanisms to evade antibiotic treatment. In addition, is able to invade and persist within human cells, hiding from the immune response and antibiotic therapies. For these reasons, novel antibacterial strategies against these pathogens are needed. Here, we developed lytic enzymes which are able to effectively target drug-resistant and intracellular Fusion of these so-called enzybiotics to cell-penetrating peptides enhanced their uptake and intracellular bactericidal activity in cell culture and in an abscess mouse model. Our results suggest that cell-penetrating enzybiotics are a promising new class of therapeutics against staphylococcal infections.
是人类健康护理中的一个主要关注点，主要是由于抗生素耐药性的日益流行。的细胞内定位在通过保护病原体免受抗生素和免疫反应的反复感染中起着关键作用。肽聚糖水解酶 (PGH) 是针对药物敏感和耐药细菌均具有高特异性杀菌作用的酶。然而，能够有效靶向细胞内的 PGH 尚未出现。为了克服这一限制，我们首先在真核细胞内室中发现的条件下筛选了 322 种重组 PGH 对葡萄球菌的裂解活性。最活跃的构建体通过融合到不同的细胞穿透肽 (CPP) 进行修饰，导致各种 菌株（包括耐甲氧西林金黄色葡萄球菌 [MRSA]）在不同的组织培养感染模型中的摄取增加和细胞内杀伤增强（多达 4.5 对数单位的减少）。协同作用的 PGH-CPP 构建体的联合应用进一步增强了它们的细胞内功效。最后，协同作用的 PGH-CPP 鸡尾酒在周围注射后，在小鼠脓肿模型中使总 减少了超过 2.2 对数单位。与单独使用 PGH 相比，PGH-CPP 杀死了更多的细胞内细菌。总的来说，我们的研究结果表明，融合 CPP 的 PGH 是针对细胞内和耐药 的有效新型蛋白治疗药物。抗生素耐药细菌的日益流行是我们这个时代最紧迫的问题之一。 是一种重要的人类病原体，已经获得了几种逃避抗生素治疗的机制。此外， 能够侵入并在人体细胞内持续存在，躲避免疫反应和抗生素治疗。出于这些原因，需要针对这些病原体的新型抗菌策略。在这里，我们开发了能够有效靶向耐药和细胞内 的裂解酶。将这些所谓的酶与细胞穿透肽融合增强了它们在细胞培养和脓肿小鼠模型中的摄取和细胞内杀菌活性。我们的结果表明，细胞穿透酶是治疗葡萄球菌感染的一种很有前途的新型治疗方法。
