Ocular Surface Microbiome, Virome, and Pathogens in Ophthalmic Disease and Health

项目摘要/摘要 这份K08申请Thuy Doan，M.D.，Ph.D.，描述了一项旨在提升她的五年战略 研究和专业技能，以成为一名独立的临床科学家为职业目标 提高对眼部感染性和炎症性疾病的认识和治疗水平。 感染性葡萄膜炎和眼部感染是全世界眼部疾病和失明的主要原因。而当 及时和准确的诊断对于改善视力结果至关重要，但明确的诊断往往不是 鉴于对易患眼部感染的因素的显著认识差距以及 可用于眼科的诊断测试。这一提议的核心假设是细菌和病毒 眼表的成分与眼睛健康和疾病有关。这个项目的目标是 了解这一潜在的病原体储存库，眼表微生物群和病毒体，并识别 与眼部感染有关的感染性物质。我们建议提高眼部分子的能力 诊断和加强对眼部感染发病机制的了解，目的如下。 第一个目标是评估基于下一代测序(NGS)的方法作为可行的临床方法的使用 眼部感染的诊断试验。这些实验将直接将NGS与当前的常规 利用存档的眼内样本进行分子诊断。预期收集的水样将在随后 接受潜在病原体的评估。第二个目标将描述抗生素对细菌的影响。 和病毒社区使用在一项大型随机对照试验中收集的样本。这些研究具有 有可能改进诊断范例并指导针对眼部感染的有针对性的干预。 我致力于在眼科领域发展以临床研究为导向的职业生涯 并了解需要进行更多的正式培训和指导。钥匙 职业发展计划的组成部分包括：(1)眼表实验室检查 微生物组、病毒组和眼部感染性疾病的病因；(2)来自多学科小组的指导 具有先进分子技术、生物统计学和临床专业知识的科学家和临床科学家 试验设计；(3)扩展分子生物学、生物统计学和临床微生物学的正规教学 知识；(4)R01应用的数据收集；和(5)向独立的计划过渡，包括 职业发展。一个科学咨询和指导委员会已经成立，以确保 走向独立。职业发展计划将在加州大学执行 加州，旧金山，这为进行这项研究提供了特殊的环境，包括州- 最先进的设施和世界级的、支持我的科学家社区，这些都对我未来的成功至关重要。

PROJECT SUMMARY/ABSTRACT This K08 application for Thuy Doan, M.D., Ph.D., describes a five year strategy designed to enhance her research and professional skills with the career goal of becoming an independent clinician scientist advancing the understanding of ocular infectious and inflammatory diseases and improving their treatment. Infectious uveitis and ocular infections are a major cause of ocular morbidity and blindness worldwide. While timely and accurate diagnosis is crucial for improved visual outcomes, a definitive diagnosis is often not possible given the significant knowledge gap in factors predisposing to eye infections and the limitations of available diagnostic tests in ophthalmology. The core hypothesis of this proposal is that the bacterial and viral constituents of the ocular surface are involved in ocular health and disease. The goal of this project is to understand this potential pathogen reservoir, the ocular surface microbiome and virome, and to identify infectious agents involved in ocular infections. We propose to advance the capabilities of ocular molecular diagnostics and enhance the understanding of the pathogenesis of ocular infections with the following aims. The first aim will evaluate the use of a next-generation sequencing (NGS)-based approach as a viable clinical diagnostic test for ocular infections. These experiments will directly compare NGS to the current conventional molecular diagnostics using archived intraocular samples. Prospectively collected aqueous samples will then be assessed for potential pathogens. The second aim will characterize the effects of antibiotics on the bacterial and viral communities using samples collected in a large randomized controlled trial. These studies have the potential to improve the diagnostic paradigm and guide targeted interventions for ocular infections. I am committed to developing a clinical research-oriented career in the field of ophthalmology at a major academic institution and understand the need to pursue additional formal training and mentorship. Key components of the career development plan include: (1) laboratory investigations of the ocular surface microbiome, virome, and causes of ocular infectious diseases; (2) mentorship from a multidisciplinary group of scientists and clinician scientists with expertise in advanced molecular techniques, biostatistics, and clinical trial designs; (3) formal didactics to expand molecular biology, biostatistics, and clinical microbiology knowledge; (4) data collection for R01 application; and (5) planned transition to independence including professional development. A scientific advisory and mentorship committee has been assembled to assure progression toward independence. The career development plan will be carried out at the University of California, San Francisco, which provides an exceptional environment to conduct this research, including state- of-the-art facilities and a world-class and supportive community of scientists all critical to my future success.