Nasal Microbial Consortia Combat Antibiotic-Resistant Bacteria

项目概要：项目3 项目3的总体目标和意义是生成基于鼻微生物群的临床前验证。 用于阻断金黄色葡萄球菌和肺炎链球菌的鼻定植的治疗剂。我们 实现这一点创新方法是在人鼻类器官上培养人鼻细菌。在一起， S.金黄色葡萄球菌和肺炎占39%的病例和29%的死亡归因于抗药性 在美国，鼻腔定植是每种细菌侵入性感染的主要来源， 它们的传播;阻断定植既减少感染又减少传播。缺乏有效的疫苗 对于s.金黄色葡萄球菌或非疫苗血清型的S.肺炎，迫切需要新的非抗生素 预防两者感染的方法。目前的治疗鼻腔定居的S。金黄色葡萄球菌依赖于 局部抗生素，并在时间上与抗生素耐药性上升相关。在这里，我们解决一个差距， 了解如何用耐致病菌的微生物群重新繁殖，以改善长期结果。多 鼻腔微生物群研究报告说，当成年人没有S.金黄色，当 孩子们没有S。肺炎。例如，良性细菌Dolosigranulum的鼻腔定植 pigrum，通常与无害的棒状杆菌，与缺乏S。小儿肺炎在 成年人，D. pigrum与S呈负相关。金黄色葡萄球菌鼻腔定植。我们的初步数据显示 菌株D. Pigrum inhibit S.金黄色葡萄球菌体外生长。此外，还研究了D.猪棒状杆菌 pseudoditheriticum对S. pneumoniae体外生长，与单独使用相比。此外，委员会认为， 增强棒杆菌D.猪生长因此，我们的首要假设是，可再生的财团， 良性人鼻细菌可赋予对S.金黄色葡萄球菌和肺炎，预防 感染和传播这些具有抗药性的威胁。此外，抗病原菌的粘膜活性溶解剂 噬菌体可以增强这一点。相互作用的网络支撑着微生物群的组成。依靠财团避免了 当添加单一菌株时可能发生的意外社区相互作用的风险， 成功地与一个理想的社区重新种群的可能性。临床试验的一个主要障碍是充分识别 在人鼻粘膜环境中赋予定殖抗性的良性鼻细菌菌株的限定聚生体 鼻上皮和促进健康，完整的上皮屏障。我们的类器官培养核心已经成功 生成人类气道类器官。因此，我们将利用鼻上皮细胞的进展来克服这一障碍。 类器官结合我们在培养人类鼻腔细菌和识别分子机制方面的专业知识 鼻内微生物群的相互作用。用我们常规使用的方法，项目1和类器官培养 核心，我们将实现两个目标：识别和表征可重复的人类鼻腔微生物聚生体， 保护鼻上皮免受S.金黄色葡萄球菌/S.肺炎和2）促进上皮屏障 完整性和阻断上皮细胞侵袭。金黄色葡萄球菌/S. pneumoniae体外培养，并确定潜在的机制。

PROJECT SUMMARY: Project 3 The overall objective and significance of Project 3 is to generate preclinical validation of nasal microbiota-based therapeutics to block nasal colonization by Staphylococcus aureus and Streptococcus pneumoniae. Our innovative approach to achieve this is to cultivate human nasal bacteria on human nasal organoids. Together, S. aureus and S. pneumoniae account for 39% of cases and 29% of deaths attributed to antibiotic-resistant bacteria in the U.S. Nasal colonization is the primary source of invasive infection by each and the reservoir for their transmission; blocking colonization reduces both infection and transmission. Lacking effective vaccines against S. aureus or nonvaccine serotypes of S. pneumoniae, there is an urgent need for new nonantibiotic approaches to prevent infections by both. Current therapies for nasal decolonization of S. aureus depend on a topical antibiotic and are temporally correlated with rising antibiotic resistance. Here, we address a gap in knowledge in how to repopulate with pathobiont-resistant microbiota to improve long-term outcomes. Multiple nasal microbiota studies report benign bacterial species present when adults are free of S. aureus and when children are free of S. pneumoniae. For example, nasal colonization by the benign bacterium Dolosigranulum pigrum, often with harmless Corynebacterium, is associated with the absence of S. pneumoniae in children. In adults, D. pigrum is inversely associated with S. aureus nasal colonization. Our preliminary data indicate many strains of D. pigrum inhibit S. aureus growth in vitro. Also, cocultivation of D. pigrum with Corynebacterium pseudodiphtheriticum robustly inhibits S. pneumoniae growth in vitro, versus either alone. Moreover, Corynebacterium enhance D. pigrum growth. Thus, our overarching hypothesis is that reproducible consortia of benign human nasal bacteria can confer colonization resistance to S. aureus and S. pneumoniae, preventing infection by and transmission of these antibiotic-resistant threats. Moreover, antipathobiont mucosal-active lytic phage may augment this. Networks of interactions underpin microbiota composition. Relying on consortia avoids the risk of unexpected community interactions that can occur when adding a single strain and increases the likelihood of successful repopulation with a desired community. A major hurdle to clinical trials is to identify fully defined consortia of benign nasal bacterial strains that confer colonization resistance in the context of human nasal epithelium and promote a healthy, intact epithelial barrier. Our Organoid Cultivation Core has successfully generated human airway organoids. Thus, we will overcome this hurdle using advances in nasal epithelial organoids combined with our expertise in cultivating human nasal bacteria and identifying molecular mechanisms of interactions within nasal microbiota. With methods routinely used by us, Project 1 and the Organoid Cultivation Core, we will achieve two aims: identify and characterize reproducible human nasal microbial consortia to 1) protect nasal epithelium from colonization by S. aureus/S. pneumoniae and 2) to promote epithelial barrier integrity and block epithelial invasion by S. aureus/S. pneumoniae in vitro, and identify underlying mechanisms.