Phenotypic, transcriptomic and pathogenic study of the first cultivated TM7 strain

第一个培养的TM7菌株的表型、转录组和致病性研究

• 批准号: 9399092
• 负责人: Batbileg Bor
• 金额: $ 0.07万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-17 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9399092
• 关键词: Actinomyces odontolyticus; Address; Adhesions; Affect; Bacteria; Biochemical; Biological Models; Candidate Disease Gene; Cell Death; Cells; Chromosome Mapping; Coculture Techniques; Disease; Epithelial; Etiology; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; Genes; Genetic; Goals; Growth; Health; Human; Human Microbiome; Human body; Immune; Immune response; Individual; Inflammatory; Inflammatory Bowel Diseases; Investigation; Knock-out; Knowledge; Lead; Link; Measures; Metabolic; Methods; Microbe; Microscopy; Molecular; Molecular Profiling; Monitor; Morphology; Mouth Diseases; Oral; Oral cavity; Parasites; Pathogenesis; Pathogenicity; Pathway interactions; Periodontal Diseases; Periodontitis; Phenotype; Physiological; Plasmids; Production; Recombinant DNA; Role; Sampling; Site; Surface; System; TNF gene; Testing; Ulcer; Virulence; base; chemokine; genetic analysis; in vivo; insight; macrophage; member; microbiota; novel; oral bacteria; prevent; public health relevance; screening; therapeutic development; tissue culture; tool; tool development; transcriptomics

 DESCRIPTION (provided by applicant): The overarching objective of this application is to achieve a detailed understanding of the first cultivated TM7 phylotype from the human oral cavity and explore its pathogenic potential. Since the initiation of the Human Microbiome Project, a tremendous diversity of bacteria has been discovered in the human body, many of which have no cultured representatives. Studying these supposedly uncultivable bacteria presents a major challenge to the field. Members of the applicant division TM7 phylum are the most prevalent yet elusive of these bacteria. TM7 has been found in various human body sites and environmental samples, and also been linked to human mucosal diseases, particularly severe and ulcerative periodontitis, vaginosis, and inflammatory bowel disease. Therefore, the impact of this study is twofold: it will increase our knowledge of uncultivable bacteria and will illuminate the pathogenic potential of TM7 members. Using novel cultivation methods, our lab recently cultured the first TM7 member (TM7x) attached to the surface of a previously uncultivated Actinomyces odontolyticus strain (XH001) from the oral cavity. TM7x was not able to grow on its own, suggesting that the presence of XH001 is crucial to TM7x's survival. In contrast, XH001 had accelerated cell death and sporulation in presence of TM7x. This type of epibiotic parasite interaction is completely novel and has never been observed in oral microbiota. Furthermore, TM7x, when associated with XH001, can prevent the induction of TNFa gene expression by macrophages, suggesting its possible role in modulating the immune response. Yet many questions remain unanswered: Why does TM7x need XH001? Does TM7x adhere to the surfaces of other oral bacteria? What are their individual and combined pathogenic potentials? Based on our initial findings, I hypothesize that physical and/or metabolic interactions between TM7x and XH001 are crucial for the growth of TM7x and have a great impact on their pathogenicity. Uncovering this interaction at the molecular level will elucidate th impact of TM7x on human health and disease. To explore these hypotheses, I will carry out four interconnected but independent aims: (1) I will first thoroughly characterize the morphological and physiological changes of TM7x and XH001 during different stages of co-culture growth. (2) I will next analyze the global gene expression of each growth stage to uncover the underlying genetics of the observed morphological changes. (3) I will manipulate gene expression of TM7x or XH001 to further evaluate the interaction between TM7x and XH001. (4) Once I understand the interaction between TM7x and XH001, I will apply this knowledge to study the pathogenic potential of TM7x in association with XH001.

 描述（由应用提供）：本应用的总体目标是详细了解人类口腔中的第一个培养的TM7系统型，并探索其致病潜力。自人类微生物组项目的倡议以来，在人体中发现了大量细菌的多样性，其中许多人没有文化的代表。研究这些预期的不可耕种细菌对该领域提出了重大挑战。适用的TM7门的成员是这些细菌中最普遍但最弹性的成员。在各种人体部位和环境样本中发现了TM7，也与人体粘膜疾病有关，尤其是严重和溃疡性牙周炎，阴道化和炎症性肠病。因此，这项研究的影响是双重的：它将增加我们对无法养殖细菌的了解，并阐明TM7成员的致病潜力。使用新的培养方法，我们的实验室最近培养了第一个TM7成员（TM7X），该TM7成员（TM7X）附着于先前未经培养的放线症脑溶液菌株（XH001）的表面。 TM7X无法自行生长，这表明XH001的存在对于TM7X的生存至关重要。相比之下，XH001在存在TM7X的情况下加速了细胞死亡和孢子形成。这种类型的表演寄生虫相互作用是完全新颖的，在口服微生物群中从未观察到。此外，当与XH001相关的TM7X可以防止巨噬细胞诱导TNFA基因表达，这表明其在调节免疫反应中的可能作用。然而，许多问题仍未解决：为什么TM7X需要XH001？ TM7X是否粘附在其他口腔细菌的表面上？他们的个体和结合的致病潜力是什么？根据我们的初步发现，我假设TM7X和XH001之间的物理和/或代谢相互作用对于TM7X的生长至关重要，并且对它们的致病性产生了很大的影响。在分子水平上发现这种相互作用将阐明TM7X对人类健康和疾病的影响。为了探讨这些假设，我将执行四个相互联系但独立的目的：（1）我首先将彻底描述TM7X和XH001在共同培养增长的不同阶段的形态和生理变化。 （2）我接下来将分析每个生长阶段的全球基因表达，以揭示观察到的形态变化的潜在遗传学。 （3）我将操纵TM7X或XH001的基因表达，以进一步评估TM7X和XH001之间的相互作用。 （4）一旦我了解TM7X和XH001之间的相互作用，我将应用这些知识来研究与XH001相关的TM7X的致病潜力。