Novel Spatial Imaging Techniques for Ocular Retinoids and Retinoid Adducts

眼部类视黄醇和类视黄醇加合物的新型空间成像技术

• 批准号: 8048066
• 负责人: Zsolt Ablonczy
• 金额: $ 17.86万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8048066
• 关键词: Address; Adult; Affect; Age; Age related macular degeneration; Aging; Americas; Area; Blindness; Blood Circulation; Carotenoids; Choroid; Collection; Complex Mixtures; Cytoplasmic Granules; Data; Development; Disease; Dissection; Employee Strikes; Environment; Eye; Fluorescence; Goals; High Pressure Liquid Chromatography; Human; Image; Imaging Techniques; Individual; Lipids; Lipofuscin; Location; Longevity; Maps; Measurement; Metabolism; Methodology; Methods; Modeling; Mus; Neurons; Normal tissue morphology; Organic solvent product; Pathogenesis; Patients; Pattern; Photoreceptors; Preparation; Prevention; Proteins; Relative (related person); Research Personnel; Resolution; Retina; Retinal; Retinal Degeneration; Retinoids; Role; Sampling; Spatial Distribution; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Structure of retinal pigment epithelium; Techniques; Technology; Tissues; Variant; Vision Disorders; Wild Type Mouse; adduct; age related; analog; base; data acquisition; design; extracellular; fetal; human tissue; lens; macula; method development; new technology; novel; novel strategies; prenatal; prevent; receptor; research study; tandem mass spectrometry; technique development; visual cycle

DESCRIPTION (provided by applicant): Age-related degenerations, including AMD, are the leading cause of vision loss and blindness in America today. One of the most striking features of the aging eye is the accumulation of autofluorescent lipofuscin granules in the macular area of the RPE. Lipofuscin is a complex mixture of compounds, many of which are bis-retinoid based. To date, over 20 bis-retinoids have been isolated and characterized from lipofuscin, A2E being the best studied. However, current methodologies do not allow for high spatial localization of these products. The assumption has been that the distribution of the products will correlate with the fluorescence of lipofuscin. We propose to develop matrix-assisted laser desorption ionization (MALDI) imaging methodologies for retinoid compounds and adducts in the RPE/choroid, which will allow both the identification of individual compounds and the determination of their spatial distribution in a single experiment. These compounds will be detectable at a resolution of 10-50 5m and relative quantization can be obtained. This imaging technique has been used for proteins, lipids and metabolites, but never for retinoids and not with retinal/choroid tissue. Our aims are 1) to develop this technology for a range of retinoids in the mouse RPE/choroid and 2) extend these studies to the more relevant human RPE/choroid. This would represent a major breakthrough in identifying the localization of bis-retinoid adducts in the RPE/choroid. Preliminary data demonstrate that A2E can be localized across the RPE/choroid at high resolution by this technique. Our findings show that in the human retina, the A2E levels do not correlate with the fluorescence attributed to lipofuscin. Imaging other bis-retinoid products and retinoid based compounds needs further development and is the focus of this project. The successful development of this technique will provide new and critical information to researchers studying approaches to the treatment and/or prevention of AMD and other retinal/RPE disorders.

描述（由申请人提供）：与年龄相关的退化，包括AMD，是当今美国视力丧失和失明的主要原因。衰老眼中最引人注目的特征之一是在RPE的黄斑区域中积累自动荧光脂肪霉素颗粒。脂肪霉素是化合物的复杂混合物，其中许多是基于双生型的。迄今为止，已经隔离了20多种双毒素，并从脂肪霉素中进行了特征，A2E是研究最佳的。但是，当前的方法论不允许对这些产品进行高空间定位。假设是产物的分布将与脂肪霉素的荧光相关。我们建议在RPE/脉络膜中开发基质辅助的激光解导电电离（MALDI）成像方法，用于类维生素类似化合物和加合物，这将允许在单个实验中识别单个化合物及其空间分布的鉴定。这些化合物将在10-50 5M的分辨率下可检测，并且可以获得相对量化。该成像技术已用于蛋白质，脂质和代谢物，但从未用于视黄素，而不是视网膜/脉络膜组织。我们的目的是1）为小鼠RPE/脉络膜中的一系列类维生素类似的类维生素的开发，并将这些研究扩展到更相关的人类RPE/脉络膜。这将代表识别RPE/脉络膜中BIS-肉变性加合物的定位方面的重大突破。初步数据表明，通过该技术，可以在高分辨率下在RPE/脉络膜上定位A2E。我们的发现表明，在人视网膜中，A2E水平与归因于脂肪霉素的荧光无关。成像其他BIS-类固醇产品和基于类维生素的化合物需要进一步开发，并且是该项目的重点。该技术的成功开发将为研究人员提供新的关键信息，以研究治疗和/或预防AMD和其他视网膜/RPE疾病的方法。