Examination of Enteric Pathogens with Multi-Omic Approaches

本提案中要研究的两种微生物，霍乱弧菌和ETEC，都是导致 在经济欠发达国家，疟疾的发病率和死亡率高达数百人， 每年有数千人死亡。此外，每种病毒都与近年来的疫情有关， 已经用基因组技术解决了。这些爆发突出了基因组的效用 测序在分析这些病原体的传播，进化和发病机制。我们将 利用大学疫苗开发中心正在进行的两项临床试验 马里兰州校园检查宿主：在用完全毒力的霍乱弧菌挑战期间的病原体相互作用 或ETEC。我们将在病原体信号和反应水平上研究这种相互作用，人类 免疫反应以及与现有微生物群的相互作用。肠道疾病的小动物模型 并不能真正代表人类发生的疾病过程，因此直接研究 在人类受试者中以受控方式进行宿主和病原体反应是一个独特的机会， 为这些相互作用提供了许多新的见解。据我们所知，这将是第一次， 将检查肠道病原体在运输过程中发生和/或选择的遗传变异 通过人类。此外，通过使用最先进的“组学技术来检查微生物群， 宏基因组和元转录组，以及免疫学研究，我们将能够开始模型， 感染过程中的相互作用。除了在人类宿主中的研究外，我们还扩展了 转录相互作用的检查到类器官模型系统的使用。既然我们意识到 人类的挑战实验是罕见的，我们希望将这种器官系统发展成为一种更好的方法。 感染研究的代表性替代模型。我们通过共培养来增加模型系统的复杂性 病原体与已鉴定的其他病原体一起在大肠杆菌中分离 研究，以及我们将扩大微生物群的研究，以共培养的物种/属确定与 挑战研究中疾病的严重程度增加。这些研究将提供一个前所未有的视角， 宿主、病原体和常驻微生物群的相互作用。

Both of the organisms to be studied in this proposal, V. cholerae and ETEC, are significant causes of morbidity and mortality due to diarrheal illness in less economically developed countries leading to hundreds of thousands of deaths each year. Additionally, each has been implicated in outbreaks in recent years that have been addressed with genomic techniques. These outbreaks have highlighted the utility of genome sequencing in the analysis of the dissemination, evolution and pathogenesis of these pathogens. We will take advantage of two ongoing clinical trials at the Center for Vaccine Development on the University of Maryland campus to examine the host:pathogen interactions during a challenge with fully virulent V. cholerae or ETEC. We will be examining this interaction at the level of pathogen signaling and response, human immune response, and interactions with the existing microbiota. Small animal models for enteric diseases are not truly representative of the disease process that occurs in humans, so the ability to directly study the host and pathogen response within human subjects in a controlled manner is a unique opportunity and will provide many novel insights into these interactions. This will be the first time, to our knowledge that an enteric pathogen will be examined for the genetic variation that occurs and/or is selected for during transit through humans. Additionally, by using the most cutting edge 'omic technologies to examine the microbiota, metagenome and metatranscriptome, as well as immunological studies, we will be able to begin to model the interaction during the infectious process. In addition to the studies within the human host, we extend the examination of the transcriptional interactions to the use of an organoid model system. Since we realize that the human challenge experiments are rare, we hope to develop this organoid system as a more representative surrogate model for infection studies. We increase the complexity of the model system by coculturing the pathogens with other pathogens that have been identified to be isolated together in diarrheal studies, as well as we will extend the microbiota studies to co-culture the species/genera identified with an increased severity of disease in the challenge studies. These studies will provide an unprecedented view into the interaction of the host, pathogen and resident microbiota.