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

描述（由申请人提供）：指导临床科学家研究职业发展奖（K 08）候选人Yan Zhang获得了医学和眼科临床学位，目前正在攻读博士学位。在加州大学伯克利分校的视觉科学课程的学位，这将在2013年春季完成。她已经在一个高知名度的期刊上发表了一篇关于后者研究的论文，还有2篇论文即将提交，这表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K 08职业奖以获得高级培训-通过研究视网膜色素上皮细胞（RPE）和骨形态发生蛋白（BMP）在眼睛生长调节和近视发展中的作用，进一步扩展她的科学知识和生物医学技术技能，以RPE为靶点进行近视眼基因治疗的相关探索性研究。在迄今为止的学习过程中，张博士接受了成功执行其拟议项目所需的许多学科的广泛培训。她建议在她申请的K奖的5年培训期间，通过额外的课程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，张博士应该为独立研究员的职业生涯做好充分的准备，成功地竞争研究基金。拟议中的研究将主要在克莉丝汀怀尔德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，并显著增加了全球眼科疾病的负担。这是眼睛长得过长的结果。在过去的几十年中，近视的患病率和严重程度在全球范围内有所上升，这刺激了对潜在机制的研究增加，这是开发有效的抗近视疗法的重要一步。以前的研究表明，早期的眼睛生长调节主要局限于眼睛本身。已知RPE是血-视网膜屏障的组成部分，在维持正常视网膜和脉络膜功能中具有关键作用。最近的研究结果，主要来自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重要作用。本研究的主要目的是探讨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，具体有3个方面：（1）研究RPE衍生的骨形态发生蛋白（BMP）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中的过表达作为近视眼基因治疗的可能性。补充实验包括使用小鸡作为近视模型的体内动物研究和使用人胎儿（hf）RPE作为模型的体外细胞培养研究，利用Zhang博士在两种模型方面的经验。体内结构和功能测量将使用先进的技术，包括高频A扫描超声检查，光谱域光学相干断层扫描（SD-OCT）和视网膜电图（ERG）。还将使用各种分子和细胞生物学技术，包括克隆、实时PCR、Western印迹、ELISA和免疫组织化学。在眼部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，也将被采用。拟议的研究将在来自加州大学旧金山分校弗朗西斯科的Jeanette Hyer博士的共同指导下进行，他是发育生物学专家，重点是鸡胚胎发生。三位资深科学家也同意担任她的项目顾问：Lawrence Rizzolo教授（耶鲁大学）和谢尔顿米勒博士（NEI/NIH），他们是RPE生理学和电生理学的专家，以及Kunxin Luo教授（加州大学伯克利分校），其研究重点是TGF-β家族，包括BMP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弗朗西斯科提供世界一流的研究环境和支持，为年轻科学家的独立研究生涯做准备和培训。

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.