Reactive ocular aldehyde in degenerative retinal diseases

退行性视网膜疾病中的反应性眼醛

基本信息

批准号：
9116177
负责人：
Akiko Maeda
金额：
$ 39.25万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9116177
关键词：
ATP-Binding Cassette Transporters Acute Age Age related macular degeneration Aging Aldehydes Amines Appearance Atherosclerosis Behavior Biological Bruch&apos s basal membrane structure CCL3 gene Cell Death Cell Survival Cells Characteristics Chronic Coculture Techniques Complement Complement component C1 Data Deposition Development Diabetes Mellitus Disease Ethanolamines Event Exhibits Exposure to Figs - dietary Generations Genetic Goals Homeostasis Human Imaging Techniques Immune Immune system In Vitro Inflammation Inflammatory Inflammatory Response Injection of therapeutic agent Knowledge Lasers Lead Light Lighting Link Lipofuscin Macular degeneration Maintenance Mediating Microglia Modeling Mus Mutation Nerve Degeneration Pathway interactions Persons Pharmacotherapy Phenotype Photoreceptors Physical condensation Production RNA Reporting Research Retina Retinal Retinal Degeneration Retinal Diseases Retinitis Pigmentosa Retinoids Retinol dehydrogenase Role Scanning Severities Solid Stargardt&apos s disease Stress Structure of retinal pigment epithelium TLR3 gene Testing Tissues Vision Visual Vitamin A Work age related chemokine cytokine cytotoxicity high risk improved in vivo in vivo imaging inhibitor/antagonist legally blind macrophage microscopic imaging mouse model novel therapeutics research study response retinal damage subretinal injection targeted treatment therapeutic target two-photon

项目摘要

DESCRIPTION (provided by applicant): Prior studies implicate the role of the reactive retinal aldehyde, all-trans-retinal (atRAL) in degenerative retinal diseases such as Stargardt's disease and age-related macular degeneration and contributions of inflammatory responses to those diseases. Although continuous generation of atRAL by the visual (retinoid) cycle is required to maintain vision, chronic exposure to atRAL may serve as an endogenous stress inducer that can cause low-grade chronic inflammation. Two characteristic features of low-grade chronic inflammation in the retina are complement deposition at Bruch's membrane and invasive immune cells into the subretinal space. Our previous studies provided the following notable findings: (1) Rdh8-/-Abca4-/- mice with delayed atRAL clearance from the retina that develop cone-rod dystrophy also display complement deposition at Bruch's membrane. (2) Subretinal accumulation of microglia/macrophages in Rdh8-/-Abca4-/- mice. (3) Tlr3-/- Rdh8-/-Abca4-/- mice displayed milder retinal degeneration with less inflammatory changes compared to Rdh8-/- Abca4-/- mice. These observations strongly indicate that retinal inflammation is caused by the biological responses to atRAL, and this inflammation greatly contributes to age-related disorders of the retina. We propose three specific aims to examine the role of atRAL in retinal inflammation that may contribute to retinal tissue damages. In Aim 1, the role of atRAL for recruitment of immune cells to the subretinal space will be examined. Retinal microglial behavior will be investigated in Rdh8-/-Abca4-/- mice after light which causes accumulation of atRAL using in vivo scanning laser ophthalmoscopic and two-photon microscopic imaging and immunohistochemical analysis. Changes in chemokine/cytokine production in the retina of Rdh8-/-Abca4-/- mice will be examined with RNA analyses. Aim 2 will test the hypothesis that CCL3 is a major chemokine that mediates recruitment of inflammatory cells to the subretinal space in retinal degeneration caused by atRAL accumulation, because our preliminary study show ~80-fold increase of CCL3 production along with increased numbers of immune cells in the subretinal space of Rdh8-/-Abca4-/- mice. Effects of CCL3 will be investigated in degenerative phenotypes of Rdh8-/-Abca4-/- mice by creating Ccl3-/-Rdh8-/-Abca4-/- mice, and by subretinal CCL3 injection to WT mice. TLR3-medicated CCL3 production will also be examined using Tlr3-/-Rdh8-/-Abca4-/- mice that showed milder retinal degeneration with less inflammatory changes. In Aim 3, we will test the hypothesis that invasive immune cells contribute to retinal tissue damage by employing macrophage-depletion in mice with genetic and pharmacologic approaches and drug treatments with microglial inhibitors to Rdh8-/- Abca4-/- mice, and by in vitro experiments using co-culture of primary RPE and retinal microglial cells. Experimental results of these interrelated aims outlined in this application will determine the rol of atRAL in retinal inflammation and degeneration, and also will aid in the development of a new therapeutic concept whether atRAL is an appropriate therapeutic target for age-related retinal disorders.

描述（由申请人提供）：先前的研究暗示了反应性视网膜醛，全反视网膜（ARTRAL）在退行性视网膜疾病中的作用，例如Stargardt的疾病和与年龄相关的黄斑变性以及对这些疾病的炎症反应的作用。尽管需要通过视觉（类维生素类动物）循环连续产生Atral才能保持视力，但慢性暴露于Atral可能会作为可能引起低级慢性炎症的内源性应激诱导剂。视网膜中低度慢性炎症的两个特征是Bruch膜上的补体沉积，并浸润性免疫细胞进入视网膜下空间。我们以前的研究提供了以下值得注意的发现：（1）RDH8 - / - ABCA4 - / - 小鼠具有延迟的视网膜清除率，该视网膜发生锥形 - 杆状营养不良症也会在Bruch的膜上显示补体沉积。（2）在RDH8 - / - ABCA4 - / - 小鼠中，小胶质细胞/巨噬细胞的视网膜下积累。（3）TLR3 - / - RDH8 - / - ABCA4 - / - 小鼠表现出温和的视网膜变性，与RDH8 - / - ABCA4 - / - 小鼠相比，炎症变化较小。这些观察结果强烈表明，视网膜炎症是由对阿特拉尔的生物学反应引起的，这种炎症极大地导致了与年龄相关的视网膜疾病。我们提出了三个特定的目的，以检查Atral在视网膜炎症中的作用，这可能导致视网膜组织损伤。在AIM 1中，将检查Atral在募集免疫细胞向视网膜下空间募集的作用。视网膜小胶质细胞行为将在rdh8 - / - abca4 - / - 小鼠中进行研究，从而导致使用体内扫描激光眼镜和两光子显微镜成像和免疫组织化学分析的体内扫描激光眼镜和两光子微观成像。 RNA分析将检查RDH8 - / - ABCA4 - / - 小鼠视网膜中趋化因子/细胞因子产生的变化。 AIM 2将检验以下假设：CCL3是一种主要的趋化因子，可将炎性细胞的募集到视网膜下空间中的视网膜下变性，因为我们的初步研究表明，CCL3产生的CCL3产生约80倍，以及RDH8-/ - ABCA4-/ - / - / - / - / - / - / - / - / - / - / - / - 小鼠的免疫细胞的数量增加。 CCL3的效果将在RDH8 - / - ABCA4 - / - 小鼠的退化表型中进行研究，通过创建CCL3 - / - RDH8 - / - ABCA4 - / - 小鼠，以及对WT小鼠的视网膜下CCL3注射。也将使用TLR3 - / - RDH8 - / - ABCA4 - / - 小鼠对TLR3中的CCL3产生进行检查，该小鼠显示出温和的视网膜变性，炎症变化较少。 In Aim 3, we will test the hypothesis that invasive immune cells contribute to retinal tissue damage by employing macrophage-depletion in mice with genetic and pharmacologic approaches and drug treatments with microglial inhibitors to Rdh8-/- Abca4-/- mice, and by in vitro experiments using co-culture of primary RPE and retinal microglial cells. 这些相互关联的目的的实验结果将确定视网膜炎症和变性的ARTAL的ROL，并且还将有助于开发新的治疗概念ARTRAL是否是与年龄相关的视网膜疾病的适当治疗靶标。