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.

项目摘要 最近的研究表明，数据表明眼微生物组存在并影响眼表 健康与疾病。尽管如此，确定了眼表微生物组的“核心”成分 很难在很大程度上是由于眼睛的生物量低，缺乏健壮的数据集，仅在接下来才责任 生成测序（NGS），以及无法显示微生物与主机之间的因果关系 生理。在此，我们概述了一个直接解决这些知识差距的建议 未来研究的基础旨在调查眼表微生物组如何影响健康和 疾病。我们已经招募了一个由临床和基本专家组成的多学科研究团队 眼科研究，低生物质部位的微生物组和免疫学。与这个团队一起，我们计划首先 表征健康的人类眼部微生物组在两个地理位置（匹兹堡和迈阿密） 使用分子技术（DNA和RNA测序）和培养物纵向（0、1周，3个月）。 尽管分子技术将为我们提供最广泛的细菌财团，但培养物将为我们提供 我们计划在坎贝尔实验室里储存可行的细菌，因此健康的人眼中的细菌 在未来的研究中可以成为社区的资源。接下来，因为通常会产生免疫力 微生物组的组件长期与主机相关联，我们计划使用 针对眼菌细菌的人免疫呼应，以区分定位细菌与瞬态 细菌可能被洗掉。为了进一步完善对眼细菌的理解，我们计划 与人眼细菌接种无细菌（GF）和特定病原体（SPF）小鼠。延长后 一段时间，我们计划将细菌从小鼠的眼睛中重新分离。仍然存在于眼上的细菌 表面将被认为具有更高的可能性是人类的眼部表面菌落体。这 这样的模型的开发将使其他人能够评估其他细菌的殖民能力 被认为是眼表微生物组的核心成分。总而言之，我们的提案的数据将是 整个社区的资源是因为我们将拥有：1）产生了一组强大的眼部微生物组 测序数据将提供有关眼微生物组特征稳定性和一致性的信息， 2）创建了从健康的人眼中获得的细菌库，3）使用人类的定义方案 测量细菌定植能力的免疫力，4）开发了一种体内动物模型，以评估如何评估 可以接种眼细菌并从眼睛中重新分离。这些资源将免费用于 社区，将充当进一步研究眼微生物组如何影响眼表的基础 健康与疾病。