Role in Myopia Development of Retinal Pigment Epithelium - A New Therapeutic Targ

简介(申请人提供)：导师临床科学家研究职业发展奖(K08)候选人张燕获得医学和眼科临床学位，目前正在加州大学伯克利分校视觉科学专业攻读博士学位，该学位将于2013年春季完成。她已经在后一项研究的一份知名期刊上发表了一篇文章，不久还将提交另外两篇论文，证明了良好的生产力和研究动力。作为学术界独立临床科学家职业生涯的下一步，张博士正在申请K08职业生涯奖以获得高级培训-通过研究视网膜色素上皮(RPE)和骨形态发生蛋白(BMPs)在眼睛生长调节和近视发育中的作用，进一步扩展她的科学知识和生物医学技能，RPE致力于近视基因治疗的相关探索性研究。在到目前为止的研究过程中，张博士接受了成功执行她提出的项目所需的许多学科的广泛培训。她建议在她正在申请的K奖的5年培训期内，通过额外的课程作业和实践培训来弥补剩余的不足。因此，在这个为期5年的职业发展奖结束时，张博士应该为成为一名独立研究员做好了充分的准备，成功地竞争了研究资金。这项拟议的研究将主要在克里斯汀·威尔索特博士的指导下进行，她是近视和眼睛生长调节领域的知名领先科学家。近视，或近视，是人类最常见的屈光不正之一，是全球眼病负担的重要组成部分。这是眼睛长得太长的结果。在过去的几十年里，近视的患病率和严重程度在全球范围内上升，刺激了对其潜在机制的更多研究，这是开发有效的抗近视疗法的关键一步。以前的研究表明，早期的眼睛生长调节在很大程度上局限于眼睛本身。众所周知，RPE是血-视网膜屏障的组成部分，在维持正常的视网膜和脉络膜功能方面起着关键作用。最近的研究发现--大部分来自Wildsoet实验室，其中大部分属于张博士--表明，它在调节眼睛生长方面也起着至关重要的作用。本研究旨在探讨RPE在近视发生发展中的作用，以及作为近视治疗的潜在靶点，目的有三个：(1)研究RPE来源的骨形态发生蛋白(BMPs)在眼生长调节中的作用；(2)研究多巴胺(DA)对RPE-BMP表达和分泌的影响；(3)探讨BMPs在RPE中的过度表达作为一种潜在的近视基因治疗方法。补充实验包括体内动物实验，使用鸡作为近视模型，以及体外细胞培养研究，使用人类胎儿(HF)RPE作为模型，利用张博士在这两个模型上的经验。体内结构和功能的测量将使用先进的技术，包括高频A超、光谱域光学相干断层扫描(SD-OCT)和视网膜电成像(ERG)。还将使用各种分子和细胞生物学技术，包括克隆、实时聚合酶链式反应、Western印迹、酶联免疫吸附试验和免疫组织化学。眼科基因治疗研究中最常见的技术，包括电穿孔和视网膜下注射，也将被使用。这项拟议的研究将在加州大学旧金山分校的珍妮特·海尔博士的共同指导下进行，海尔博士是发育生物学专家，重点是鸡的胚胎发生。三位资深科学家也同意担任她项目的顾问：耶鲁大学的Lawrence Rizzolo教授和Nei/NIH的Sheldon Miller博士，他们是视网膜色素上皮生理学和电生理学方面的专家；以及加州大学伯克利分校的罗坤欣教授，他的研究重点是转化生长因子-�家族，包括骨形态发生蛋白及其在细胞分化、组织形态发生和细胞外基质产生中的作用。加州大学伯克利分校和加州大学旧金山分校提供世界一流的研究环境，支持年轻科学家为独立研究事业做准备和培训。

DESCRIPTION (provided by applicant): The Mentored Clinical Scientist Research Career Development Award (K08) candidate, Yan Zhang, earned her clinical degrees in Medicine and Ophthalmology, and she is currently pursuing her Ph.D. degree in the Vision Science Program at University of California, Berkeley, which will be completed in the Spring 2013. She has already one publication in a high profile journal from the latter research, with 2 more papers soon to be submitted, demonstrating both good productivity and research drive. As the next step towards a career as an independent clinician scientist in academia, Dr. Zhang is applying for a K08 career award to obtain advanced training - to further expand her scientific knowledge and biomedical technical skills through investigations into the roles of retinal pigment epithelium (RPE) and bone morphogenetic proteins (BMPs) in eye growth regulation and myopia development, with the RPE targeted in related exploratory studies of gene therapy for myopia treatment. Over the course of her studies to-date, Dr. Zhang has received broad training in many of the disciplines required for successfully executing her proposed project. She proposed to cover remaining deficiencies with additional coursework and hands-on training during the 5-year training period of the K award for which she is applying. Thus at the end of this 5-year career development award Dr. Zhang should be well prepared for a career as an independent researcher, successfully competing for research funds. The proposed study will be conducted primarily under the mentorship of Dr. Christine Wildsoet, who is a well- known leading scientist in the field of myopia and eye growth regulation. Myopia, or nearsightedness, is one of most common refractive errors in humans and significantly contributes to the global burden of eye disease. It is the product of eyes growing excessively long. The prevalence and the severity of myopia have risen worldwide during the past several decades, stimulating increased research into the underlying mechanisms, an essential step in developing effective anti-myopia therapies. Previous studies have suggested that early eye growth regulation is largely localized to eye itself. The RPE is known to be a component of the blood-retina barrier, with critical roles in maintaining normal retinal and choroidal functions. Recent research findings, mostly from the Wildsoet lab and much of it belongs to Dr. Zhang, suggest that it also plays an essential role in the regulation of eye growth. The proposal focuses on the role of RPE in myopia development and as a potential target for myopia therapy, with 3 specific aims: (1) to investigate the role of RPE-derived bone morphogenetic proteins (BMPs) in eye growth regulation; (2) to investigate the effects of dopamine (DA) on RPE-BMP expression and secretion; (3) to investigate over-expression of BMPs in RPE as a potential gene therapy for myopia. Complementary experiments include in vivo animal studies, using the chick as an myopia model, and in vitro cell culture studies, using human fetal (hf) RPE as a model, exploiting Dr. Zhang's experience with both models. In vivo structural and functional measurements will use advanced technologies, including high frequency A-scan ultrasonography, spectral domain optical coherence tomography (SD-OCT) and electroretinography (ERG). A variety of molecular and cell biology techniques will also be used including cloning, real-time PCR, Western blot, ELISA, and immunohistochemistry. Techniques most commonly encountered in ocular gene therapy research, including electroporation and subretinal injection, will also be employed. The proposed research will be conducted under the co-mentorship of Dr. Jeanette Hyer from UC San Francisco, who is an expert in developmental biology, with a focus on chick embryogenesis. Three senior scientists have also agreed to serve as consultants on her project: Professor Lawrence Rizzolo (Yale University) and Dr. Sheldon Miller (NEI/NIH), who are experts in RPE physiology and electrophysiology, and Professor Kunxin Luo (UC Berkeley), whose research focus is the TGF-� family including BMPs and their roles in cell differentiation, tissue morphogenesis, and extracellular matrix production. UC Berkeley and UC San Francisco offer world-class research environments and support for the preparation and training of young scientists for independent research careers.