Defining bacterial members of the ocular surface microbiome and assessing stability over time

定义眼表微生物组的细菌成员并评估随时间推移的稳定性

• 批准号: 10668753
• 负责人: Anat Galor
• 金额: $ 111.72万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10668753
• 关键词: Address; Age; Animal Model; Bacteria; Bacterial DNA; Biomass; Cells; Clinical; Collection; Communities; Contact Lenses; DNA; DNA sequencing; Data; Data Set; Development; Disease; Equilibrium; Eye; Eye diseases; Flow Cytometry; Foundations; Future; Geographic Locations; Geography; Germ-Free; Gnotobiotic; Goals; Health; Human; Immune; Immune response; Immune system; Immunity; Immunologic Stimulation; Immunology; In Vitro; Incubated; Individual; Infection; Infectious Agent; Integration Host Factors; Interdisciplinary Study; Knowledge; Laboratories; Link; Measures; Microbe; Microbiology; Modeling; Molecular; Monitor; Mus; Nature; Ophthalmology; Organism; Pathology; Patients; Peripheral Blood Mononuclear Cell; Phenotype; Physiological; Physiology; Positioning Attribute; Predisposition; Process; Production; Proliferating; Protocols documentation; RNA analysis; Research; Resolution; Resources; Role; Site; Swab; Symptoms; System; Techniques; Testing; Time; Ulcerative Colitis; base; cost; cytokine; design; examination questions; experience; fecal transplantation; geographic difference; germ free condition; gut microbiome; host microbiome; human subject; in vivo; longitudinal analysis; member; metagenomic sequencing; microbiome; microbiome analysis; microbiome components; microbiome composition; microbiome research; microbiome sequencing; microbiome signature; mouse model; next generation sequencing; ocular microbiome; ocular surface; ocular surface disease; programs; recruit; repository; response; sex; transcriptome sequencing

PROJECT SUMMARY Recent studies have shown data suggesting that the ocular microbiome exists and influences ocular surface health and disease. Despite this defining the “core” components of the ocular surface microbiome has been difficult largely due to the low biomass nature of the eye, a lack of robust data sets, reliance only on next generation sequencing (NGS), and the inability to show causal relationships between microbes and host physiology. Herein, we have outlined a proposal that directly addresses these knowledge gaps and that will lay the foundation for future studies aimed at investigating how the ocular surface microbiome influences health and disease. We have recruited a multidisciplinary research team that consists of experts in clinical and basic ophthalmology research, microbiome of low biomass sites, and immunology. With this team, we plan to first characterize the healthy human ocular microbiome in two geographic locations (Pittsburgh and Miami) longitudinally (0, 1 week, 3 months) using molecular techniques (DNA and RNA sequencing) and culturomics. While molecular techniques will provide us with the broadest consortia of bacteria, culturomics will provide us with viable bacteria that we plan to bank in the Campbell Laboratory, so that bacteria from healthy human eyes can be a resource for the community in future studies. Next, because immunity is normally generated towards components of the microbiome that remain associated with the host for extended periods of time, we plan to use human immune responses directed against ocular bacteria to distinguish colonizing bacteria from transient bacteria that are likely washed away. To further refine the understanding of ocular colonizing bacteria, we plan to inoculate germ free (GF) and specific pathogen free (SPF) mice with human ocular bacteria. After an extended period of time, we plan to re-isolate bacteria from the eyes of mice. Bacteria that are still present on the ocular surface will be considered to have a higher likelihood of being an ocular surface colonizer in humans. The development of a model such as this will allow others to assess the colonizing ability of other bacteria that may be considered a core component of the ocular surface microbiome. In sum, data from our proposal will be a resource for the community at large because we will have: 1) generated a robust set of ocular microbiome sequencing data that will provide information on the stability and consistency of ocular microbiome signatures, 2) created a bank of bacteria that were acquired from healthy human eyes, 3) defined protocols to use human immunity to measure the colonizing ability of bacteria, 4) developed an in vivo animal model to assess how ocular bacteria can be inoculated and re-isolated from the eye. These resources will be free to use for the community and will act as a base to further investigate how the ocular microbiome influences ocular surface health and disease.

项目概要 最近的研究显示数据表明眼部微生物组的存在并影响眼表 尽管如此，眼表微生物组的“核心”组成部分一直被定义为健康和疾病。 困难主要是由于眼睛的生物量低、缺乏可靠的数据集、仅依赖于下一个 世代测序（NGS），以及无法显示微生物和宿主之间的因果关系 在此，我们概述了一项直接解决这些知识差距的提案，该提案将奠定基础。 为未来研究奠定基础，旨在调查眼表微生物组如何影响健康和 我们招募了由临床和基础专家组成的多学科研究团队。 我们计划首先在眼科研究、低生物量位点的微生物组和免疫学方面进行研究。 描述两个地理位置（匹兹堡和迈阿密）健康人类眼部微生物组的特征 使用分子技术（DNA 和 RNA 测序）和培养组学进行纵向（0、1 周、3 个月）分析。 分子技术将为我们提供最广泛的细菌群落，而培养组学将为我们提供 与我们计划在坎贝尔实验室储存的活细菌一起使用，以便来自健康人眼的细菌 接下来，可以成为社区未来研究的资源，因为免疫力通常是针对以下目标产生的。 微生物组的组成部分与宿主长时间保持联系，我们计划使用 针对眼部细菌的人类免疫反应，以区分定植细菌和短暂细菌 为了进一步加深对眼部定植细菌的了解，我们计划。 在长时间后给无菌（GF）和无特定病原体（SPF）小鼠接种人类眼部细菌。 一段时间后，我们计划从小鼠眼中重新分离出仍然存在于眼部的细菌。 表面将被认为更有可能成为人类眼表定植者。 开发这样的模型将允许其他人评估其他细菌的定殖能力，这些细菌可能 被认为是眼表微生物组的核心组成部分总而言之，我们提案中的数据将是 为整个社区提供资源，因为我们将：1）生成一组强大的眼部微生物组 测序数据将提供有关眼部微生物组特征的稳定性和一致性的信息， 2) 创建了从健康人眼中获取的细菌库，3) 定义了使用人类眼睛的方案 免疫来测量细菌的定植能力，4) 开发了体内动物模型来评估如何 可以从眼睛中接种并重新分离出眼部细菌，这些资源将免费供患者使用。 社区并将作为进一步研究眼部微生物组如何影响眼表的基础 健康和疾病。