Combining Bacteriophage-Mediated Decolonization with Microbial Engraftment to Foster Sustained Supragingival Microbiome Modulation for Enhanced Anti-Cariogenic Therapy

摘要 Vulcan Biologics正在开发一个平台，该平台利用合成生物学的力量来实现持久和 稳定调节口腔微生物群，预防龋齿。龋齿的特征是 膳食糖与牙科生物膜接触时牙釉质的酸化和降解 (斑块)。龋齿是人类最常见的慢性病，影响42%的儿童，59%的儿童 青少年和92%的美国成年人。它可能伴随着严重的并发症和并发症， 包括疼痛、脓肿、牙科败血症，甚至死亡。龋齿还会导致生活质量下降和 据估计，每年造成的生产力损失达54亿美元。目前预防龋齿的方法包括 刷牙/使用牙线和使用氟化物对降低儿童龋齿的总体患病率几乎没有帮助。 在过去十年中，人口数量不断增加，因此有必要制定更有效的措施。有一个 提高对口腔微生物群在龋病发病机制中作用的认识。变形链球菌是 口腔微生物区系中自然产生的成员，是人类龋齿的主要病原体。 Vulcan Biologics提出了一种三方方法来持续和稳定地重新编程口腔 微生物组以减少或消除变形链球菌的流行。此方法基于将 噬菌体--仅针对细菌但不影响真核细胞的病毒--的应用 提供益生菌效应的选定细菌和保护性益生菌所特有的营养物质(益生素 细菌。这一非殖民化/再殖民化(非殖民化-再殖民化)战略有可能在以前的 为微生物组提供有效和持久的调节的尝试失败了，即在克服 已建立的微生物群落对新来者永久植入的抵抗力。为了证明 这种方法的可行性，在这个第一阶段的项目中，火神生物公司将：1)建立和表征细菌 以及噬菌体文库，包括从唾液样本中分离新的毒力噬菌体并分析其宿主 2)用抗菌肽和生物膜降解酶改造强毒噬菌体，以提高S. 3)建立生物被膜模型，评价噬菌体对变形链球菌的杀灭作用 单独治疗，单独益生菌治疗，或它们的组合。这将考验前两种方法的有效性。 我们的三部分方法中的步骤，并为第二阶段工作铺平道路，以评估整合所有 3步(即提供益生素)和在大鼠模型中。归根结底，这种方法有望提供一个长期的 和持之以恒的防龋法。

Abstract Vulcan Biologics is developing a platform that harnesses the power of synthetic biology to enable persistent and stable modulation of the oral microbiome for the prevention of dental caries. Caries are characterized by the acidification and degradation of tooth enamel when dietary sugars come into contact with dental biofilms (plaque). Dental caries is the most prevalent chronic disease in humans, affecting 42% of children, 59% of adolescents, and 92% of adults in the U.S. It can be accompanied by serious comorbidities and complications, including pain, abscesses, dental sepsis, and even death. Caries also contribute to diminished quality of life and account for an estimated $5.4 billion in lost productivity each year. Current methods to prevent caries, including brushing/flossing and fluoride applications, have done little to reduce the overall prevalence of caries in the population in the last decade, thus necessitating the development of more effective measures. There is an increasing awareness of the role of the oral microbiome in dental caries pathogenesis. Streptococcus mutans is a naturally occurring member of the oral microbiota and the principal etiological agent of dental decay in humans. Vulcan Biologics proposes a tripartite approach for the persistent and stable reprogramming of the oral microbiome to reduce or eliminate the prevalence of S. mutans. This approach is based on combining the application of bacteriophages—viruses that exclusively target bacteria but do not affect eukaryotic cells—with selected bacteria providing a probiotic effect and nutrients (prebiotics) that are specific to the protective probiotic bacteria. This decolonization/recolonization (Decon-Recon) strategy has the potential to succeed where previous attempts to provide effective and enduring modulation of the microbiome have failed, namely in overcoming the resistance of established microbial communities to the permanent engraftment of newcomers. To demonstrate the feasibility of this approach, in this Phase I project, Vulcan Biologics will: 1) establish and characterize bacterial and phage libraries, including isolating novel virulent phages from saliva samples and analyzing their host ranges; 2) engineer virulent phages with antimicrobial peptides and biofilm-degrading enzymes to improve S. mutans killing ability; and 3) establish a biofilm model and assess S. mutans reduction resulting from phage treatment alone, probiotic treatment alone, or their combination. This will test the effectiveness of the first two steps in our 3-part approach and pave the way for Phase II work to evaluate the efficacy upon integration of all 3 steps (i.e., providing a prebiotic) and in a rat model. Ultimately, this approach promises to provide a long-term and persistent method to prevent dental caries.