Metagenomic Analysis of Ocular Surface Microbiome

描述（由申请人提供）：微生物侵入眼表（OS）损害角膜清晰度并引起炎症，如角膜炎、眼内炎和沙眼等致盲疾病。然而，在许多情况下，病原体仍然是未知的，因为它不能在培养中生长。在这个项目中，我们将利用最新的DNA测序技术来克服这一限制。我们的最终目标是研究人类OS的共生微生物种群在健康和疾病中的作用。我们的方法是使用“培养独立”技术来检测和分配非人类DNA到分类学上不同的微生物物种。同样的技术将与生物信息学相结合，用于将微生物基因注释到功能途径，以确定微生物种群在操作系统上的潜在活性。我们将分析健康结膜和角膜样本，以检查眼表不同区室中微生物群落（“微生物组”）的差异。我们提出了巴斯科姆帕尔默眼科研究所的多学科团队和阿贡国家实验室（Argonne， IL）的微生物基因组学专家的协调努力，首次调查了OS微生物组。在这个项目中获得的新知识有望彻底改变对眼部微生物生态的理解，并促进新型诊断工具的设计。该项目的成功完成将推动该领域朝着测试操作OS微生物组的治疗潜力的方向发展。

DESCRIPTION (provided by applicant): Microbial invasion into the ocular surface (OS) compromises corneal clarity and causes inflammation in blinding conditions like keratitis, endoophthalmitis and trachoma. However, in many cases the pathogen(s) remains unidentified because it fails to grow in culture. In this project we will utilize latest DNA sequencing technology to overcome this limitation. Our ultimate goal is to investigate the role of commensal microbial population of the human OS in health and disease. Our approach is to use the "culture-independent" technology that detects and assigns non-human DNA to taxonomically distinct microbial species. The same technology in combination with bioinformatics will be utilized to annotate microbial genes to functional pathways, in order to determine potential activity of microbial population on the OS. We will analyze samples of healthy conjunctiva and corneas to examine differences in microbial communities ("microbiome") in different compartments of ocular surface. We put forth a coordinated effort of multidisciplinary team at Bascom Palmer Eye Institute and microbial genomics experts at the Argonne National Laboratory (Argonne, IL) to investigate, for the first time, the OS microbiome. The new knowledge acquired in this project is expected to revolutionize the understanding of ocular ocular microbial ecology, and facilitate designing of novel diagnostic tools. Successful completion of this project will advance the field towards testing the therapeutic potential of manipulation the OS microbiome. PUBLIC HEALTH RELEVANCE: The ultimate goal of this new project is recruit new sequencing technology to detect, identify and catalogue all microorganisms that populate human ocular surface, and can potentially affect vision. We will undertake the first comprehensive characterization of microbial diversity of healthy ocular surface that will allow us to monitor bacterial diversity in healthy and diseased eyes. Successful completion of this project will bring to a new level both diagnostics and understanding of ocular infections. PHS 398/2590 (Rev. 09/04, Reissued 4/2006) Page Continuation Format Page