A New Imaging Method for Ocular Lens Protein Modifications

描述(申请人提供)：该眼镜片为蛋白质生物化学提供了独特的环境，这是在生物体的整个生命周期中保持透明度所必需的。细胞器丢失后，蛋白质的周转很少，允许翻译后的修饰在生物体的整个生命周期中积累。正是这些修饰与蛋白质聚集、不溶解和白内障的形成有关。虽然已经对一些修饰进行了分类，但对修饰蛋白在人类晶状体中的空间分布研究很少。正是修改的空间模式可以区分正常的发育或调节修改与年龄相关的修改和特定于白内障的修改。该方案的目的是开发一种新的成像技术，用于测量高空间分辨率的晶状体蛋白质分布。MALDI组织成像已被应用于几种组织，以绘制肿瘤特异性蛋白、器官特异性蛋白分布以及感染组织和正常组织中的分布。成像晶状体蛋白质及其修饰的前景是，白内障发生的新机制将被揭示。事实上，在初步研究中，有两个以前没有报道的修饰已经直接从晶状体组织成像。这一开发应用的第一个目标是确定成像眼晶状体蛋白质的最佳样品制备和图像采集要求。第二个目的是将目标1中确定的条件应用于不同年龄和白内障状况的人类晶状体成像。最终，将获得关于晶状体蛋白修饰的新的空间分辨信息，这些信息可用于形成关于白内障形成的假说，并导致延迟安抚开始的策略。与公共健康相关：建议开发一种新的眼晶状体组织成像技术，在该技术中将获得年龄和白内障特异性蛋白质修饰的空间分布。通过这项新技术实现的晶状体蛋白质修饰的空间模式的知识将导致新的机制假说和潜在的白内障治疗目标。

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.