A New Imaging Method for Ocular Lens Protein Modifications

描述（由申请人提供）：眼用透镜为蛋白质生物化学提供了一个独特的环境，这是在生物体的整个生命周期内保持透明度的需要所必需的。在细胞器丧失后，很少发生蛋白质周转，允许翻译后修饰在生物体的一生中积累。正是这些修饰与蛋白质聚集、不溶解和白内障形成有关。虽然已经对许多修饰进行了编目，但是对修饰蛋白在人类透镜中的空间分布的研究很少。正是修饰的空间模式可以将正常发育或调节修饰与年龄相关和白内障特异性修饰区分开来。本提案的目的是开发一种新的成像技术，用于测量具有高空间分辨率的透镜蛋白质分布。MALDI组织成像已应用于几种组织，以绘制肿瘤特异性蛋白质、器官特异性蛋白质分布以及感染组织与正常组织中的分布。对透镜蛋白质及其修饰进行成像的前景是将揭示白内障发生的新机制。事实上，在初步研究中，已经直接从透镜组织成像了两种以前未报告的修改。该开发应用的第一个目的是确定用于成像眼透镜蛋白的最佳样品制备和图像采集要求。第二个目标是应用目标1中确定的条件对不同年龄和白内障状态的人晶状体进行成像。最终，新的空间分辨信息透镜蛋白质修饰将被收购，可用于制定白内障形成的假设，并导致策略，以延迟发病的安抚。公共卫生相关性：提出了一种新的成像技术的发展，用于眼透镜组织中的年龄和白内障特异性蛋白质修饰的空间分布将被收购。通过这种新技术实现的透镜蛋白质修饰的空间模式的知识将导致新的机制假说和白内障的潜在治疗靶点。

DESCRIPTION (provided by applicant): The ocular lens provides a unique environment for protein biochemistry necessitated by the need to maintain transparency over the lifetime of the organism. After organelle loss, little protein turnover occurs allowing post-translational modifications to accumulate over the lifetime of the organism. It is these modifications that have been linked to protein aggregation, insolubilization, and cataract-formation. Although a number of modifications have been cataloged, the spatial distribution of modified proteins in the human lens has been little-studied. It is precisely the spatial patterns of modification that can differentiate normal developmental or regulatory modifications from age-related and cataract-specific modifications. The objective of this proposal is to develop a new imaging technology for measuring lens protein distributions with high spatial resolution. MALDI tissue imaging has been applied to several tissues to map tumor-specific proteins, organ-specific protein distributions, and distributions in infected versus normal tissues. The promise of imaging lens proteins and their modifications is that new mechanisms of cataractogenesis will be revealed. Indeed, in preliminary studies, two previously unreported modifications have been imaged directly from lens tissue. The first aim of this developmental application is to determine optimum sample preparation and image acquisition requirements for imaging ocular lens proteins. The second aim is apply the conditions determined in aim 1 to image human lenses of varying age and cataract status. Ultimately, new spatially-resolved information on lens protein modification will be acquired that can be used to formulate hypotheses on cataract formation and lead to strategies to delay the onset of pacification. PUBLIC HEALTH RELEVANCE: The development of a new imaging technology is proposed for ocular lens tissue in which the spatial distributions of age- and cataract-specific protein modifications will be acquired. Knowledge of spatial patterns of lens protein modifications achieved by this new technology will lead to new mechanistic hypotheses and potential therapeutic targets for cataracts.