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，利用合成生物学的力量， 稳定调节口腔微生物组以预防龋齿。龋齿的特征是 当膳食糖与牙生物膜接触时牙釉质的酸化和降解 （饰板）。龋齿是人类最普遍的慢性疾病，影响42%的儿童，59%的儿童， 在美国，青少年和92%的成年人可能伴有严重的合并症和并发症， 包括疼痛，呕吐，牙脓毒症，甚至死亡龋齿也会降低生活质量， 据估计，每年损失的生产力为54亿美元。目前预防龋齿的方法，包括 刷牙/牙线和氟化物的应用，几乎没有减少龋齿的总体患病率， 因此，有必要制定更有效的措施。有一个 提高对口腔微生物组在龋齿发病机制中作用的认识。变形链球菌是 口腔微生物群中自然存在的成员，是人类龋齿的主要病原体。 Vulcan Biologics提出了一种三方方法，用于持久和稳定的口腔 微生物组，以减少或消除S.变异人这种方法基于将 噬菌体--专门靶向细菌但不影响真核细胞的病毒--的应用， 选择的提供益生菌作用的细菌和对保护性益生菌特异的营养物（益生元 细菌这一非殖民化/非殖民化战略有可能在以前的 提供微生物组的有效和持久调节的尝试失败了，即在克服微生物组的缺陷方面失败了。 已建立的微生物群落对新生物永久植入的抵抗力。证明 为了验证这种方法可行性，在第一阶段项目中，Vulcan Biologics将：1）建立和表征细菌 和噬菌体文库，包括从唾液样本中分离新的毒性噬菌体，并分析其宿主 范围; 2）用抗菌肽和生物膜降解酶工程改造毒力菌株以改善S. 3）建立生物被膜模型，评价S.噬菌体引起的变异体减少 单独治疗、单独益生菌治疗或它们的组合。这将检验前两者的有效性 在我们的三部分方法的步骤，并铺平了道路，第二阶段的工作，以评估疗效后，整合所有 3个步骤（即，提供益生元）和大鼠模型中。最终，这种方法有望提供长期的 和持久的方法来预防龋齿。

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.