Host tRNA-derived small RNAs (tsRNAs) mediate interactions between host and oral microbes

宿主 tRNA 衍生的小 RNA (tsRNA) 介导宿主和口腔微生物之间的相互作用

• 批准号: 10446416
• 负责人: Xuesong He
• 金额: $ 47.84万
• 依托单位: ADA FORSYTH INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-22 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10446416
• 关键词: Address; Affect; Bacteria; Bacterial Genes; Bacterial Proteins; Behavior; Biochemical Genetics; Biochemistry; Biological Models; Biological Process; Carrier Proteins; Cell Line; Cells; Communication Programs; Complex; Coupled; Data; Defense Mechanisms; Disease; Dissection; Encapsulated; Epithelial Cells; Exhibits; Fluorescence Microscopy; Follow-Up Studies; Fusobacterium nucleatum; Gene Expression; Genes; Goals; Growth; Health; Homeostasis; Host Defense; Human; Image; Infectious Agent; Intake; Knowledge; Light; Maintenance; Mediating; Membrane; Membrane Transport Proteins; Microbe; Mouth Diseases; Mucous Membrane; National Institute of Dental and Craniofacial Research; Natural Immunity; Oral; Oral health; Oral mucous membrane structure; Pb in saliva; Periodontitis; Plants; Play; Porphyromonas; Process; Protein Binding Domain; Protocols documentation; Publishing; RNA; RNA Recognition Motif; Regulatory Element; Regulatory Pathway; Reporting; Research; Resolution; Role; Saliva; Salivary; Small RNA; Strategic vision; Streptococcus mitis; Surface; Testing; Therapeutic; Transfer RNA; United States National Institutes of Health; Work; adaptive immunity; antimicrobial; bacterial genetics; base; beneficial microorganism; cell immortalization; exosome; extracellular; genetic analysis; gut microbiome; host microbiome; host-microbe interactions; human subject; keratinocyte; microbial; microbial host; microbiome; novel therapeutic intervention; nucleic acid-based therapeutics; opportunistic pathogen; oral bacteria; oral commensal; oral microbiome; pathogen; periodontopathogen; prevent; response; saliva sample; social; trafficking; transcriptome sequencing

Host mucosal surfaces are highly specialized and possess a complex array of innate and adaptive immunity. They provide the first line of protection against infectious agents by initiating protective responses to potential pathogens. Furthermore, the symbiotic relationship of the hundreds of microbial species with the host requires a fine-tuned response at the mucosal surface that prevents overgrowth of opportunistic pathogens, but sparing beneficial microbes. While small RNAs (sRNAs) have been regarded as a class of intracellular regulatory elements, emerging studies in plant-pathogen and host-gut microbiome interactions uncover that hosts can exploit sRNAs, such as transfer RNA (tRNA)-derived sRNAs (termed “tsRNAs”), via encapsulation in exosomes, as a new mode of bacterial gene modulation or defense mechanism. Yet, it remains to be elucidated whether such sRNA-mediated inter-kingdom gene modulation or defense mechanism exist in the context of host-oral microbiome interaction. To address this knowledge gap, the present R01 application will focus on host-derived, exosome-borne, salivary tsRNAs, of which the biological functions have remained largely elusive to date. Speficically, we recently demonstrated that human Normal Oral Keratinocyte- Spontaneously Immortalized (NOKSI) cells released two exosome-borne tsRNAs, tsRNA-000794 and tsRNA- 020498, when challenged with Fusobacterium nucleatum (Fn), a key oral commensal and opportunistic periodontal pathogen. Importantly, these two tsRNAs can be readily detected from saliva in healthy human subjects. Intriguingly, both tsRNAs exhibit highly selective, Fn-targeting antimicrobial activity—directly adding synthetic mimics of these two tsRNAs, but not scramble RNA, to bacterial culture inhibits the growth of Fn, but not that of Porphyromonas ginigivalis (Pg), a gram-negative periodontal pathogen or Streptococcus mitis (Sm), a health-associated oral bacterium. In preliminary work, we further took a multi-facet approach to identify a putative RNA-specific membrane transporter as well as candidate intracellular bacterial protein targets of Fn- targeting tsRNAs. Building on our comprehensive preliminary data, the goal of this application is two-fold: (Part I) achieve mechanistic understanding of the cross-kingdom trafficking of host-derived Fn-targeting tsRNAs and their modulating effect on Fn growth during NOKSI-Fn interaction, through exosome tracking and in-depth dissection of the tsRNAs transporter and intracellular targets in Fn; (Part II) expand our work to profile and compare salivary tsRNAs between healthy and periodontitis subjects, with a focus on demonstrating the broad implication of host-generated tsRNAs as a conserved mechanism to achieve host-microbial homeostasis. The realization of this application will not only address a fundamental question by defining tsRNAs as a new class of host defense molecules in maintaining host-microbiome homeostasis via targeted microbial modulation, but also will pave the way for a new therapeutic strategy against oral diseases, considering the already successful trajectory of nucleic acid therapeutics in recent years.

宿主粘膜表面是高度专业化的，拥有一系列先天和适应性免疫学。 他们通过对潜在的保护作用提供了对感染者的第一道保护 病原体。此外，数百种微生物物种与宿主的共生关系需要 粘膜表面的微调反应可防止机会病原体的过度生长，但保留 有益的微生物。而小的RNA（SRNA）被认为是一类细胞内调节 元素，植物病原体的新兴研究和宿主的微生物组相互作用发现宿主可以 通过封装在 外泌体，作为细菌基因调节或防御机制的一种新模式。然而，这仍然是 阐明了这种SRNA介导的基因调制或防御机制是否存在于 宿主 - 口腔微生物组相互作用的上下文。为了解决此知识差距，当前的R01应用程序将 专注于宿主衍生的外泌体传播，唾液TSRNA，生物学功能仍然存在 在很大程度上难以捉摸。从事方面，我们最近证明了人类正常口服角质形成细胞 - 自发永生（NOKSI）细胞释放了两个外泌体传播TSRNA，TSRNA-000794和TSRNA- 020498，当受到核细菌核（FN）的挑战时（FN），这是一个关键的口头和机会主义 牙周病原体。重要的是，这两个TSRNA可以很容易地从健康人的唾液中检测到 主题。有趣的是，这两种TSRNA都暴露了高度选择性，靶向FN的抗菌活性 - 直接添加 这两个TSRNA的合成模拟物，而不是争夺RNA，以抑制FN的生长，但 不是卟啉单胞菌（PG），革兰氏阴性牙周病原体或链球菌（SM）， 与健康相关的口腔细菌。在初步工作中，我们进一步采取了多方面的方法来识别 假定的RNA特异性膜转运蛋白以及候选FN-的细胞内细菌蛋白靶标 靶向TSRNA。在我们综合的初步数据的基础上，此应用程序的目标是两个方面：（部分 i）对托管tsrnas和 它们通过外泌体跟踪和深度调节对NOKSI-FN相互作用期间FN增长的调节作用 FN中TSRNA转运蛋白和细胞内靶标的解剖； （第二部分）将我们的工作扩展到个人资料和 比较健康和牙周炎受试者之间的唾液TSRNA，重点是证明广泛的 宿主生成的TSRNA作为实现宿主微生物稳态的组成机制的意义。这 实现此应用程序不仅将通过将TSRNA定义为新类来解决一个基本问题 通过靶向微生物调节来维持宿主微生物体稳态的宿主防御分子，但 考虑到已经成功的，还将为针对口腔疾病的新治疗策略铺平道路 近年来核酸治疗的轨迹。