TRP-CHANNELS IN LENS DEVELOPMENT AND CATARACT

TRP 通道在晶状体发育和白内障中的作用

基本信息

批准号：
10557158
负责人：
Alan Shiels
金额：
$ 38.09万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-02-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10557158
关键词：
AA Spectrophotometry Actins Address Agonist Alternative Splicing Anterior Arizona Bioinformatics Calcium Calcium Channel Cataract Cations Cell Line Cells Ciliary Body Clinical Collaborations Complement Cytoplasm Defect Deposition Development Disease Epithelial Cells Epithelium Eye Eye Development Eye diseases Functional disorder Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Genetic study Genome Glaucoma Goals Growth Homeostasis Human Image Imaging Techniques Immune response Immunofluorescence Microscopy Impairment Inherited Investigation Iris Knock-in Mouse Knockout Mice Knowledge Lens development Light Measures Membrane Messenger RNA MicroRNAs Molecular Mus Mutate Mutation Operative Surgical Procedures Optics Osmosis Pain Perception Physiological Processes Physiology Play Property Protein Isoforms RNA RNA Splicing Research Risk Factors Role Scientific Advances and Accomplishments Signal Pathway Smooth Muscle Steroids Stimulus Stress Symptoms TRP channel Techniques Technology Temperature Testing Therapeutic Tissues Transcript Transfection Universities Variant Visual impairment Water age related antagonist calcium phosphate childhood cataract deep sequencing epithelial to mesenchymal transition imaging facilities insight interdisciplinary approach lens lens morphogenesis lens transparency live cell imaging member mutant novel pressure preventive intervention receptor recruit response transcription factor transcriptome sequencing vision development

项目摘要

Project Summary/Abstract The transient receptor potential (TRP) superfamily of non-selective cation channels play critical roles in diverse physiological processes including the perception of light, temperature, pressure, and pain. In previous genetic studies, we have discovered that mutation of a member of the melastatin-related TRP-channels (TRPM3) underlies an inherited form of pediatric cataract that is variably associated with glaucoma and anterior segment defects. In this proposal, we will investigate the role of TRPM3 in lens development and cataract pathobiology using gene-edited mice and cell-lines by addressing three specific aims. In aim-1, we will characterize lens TRP-channel transcript splice-variants by means of RNA deep-sequencing and the calcium channel properties of lens TRPM3 isoforms using fluorometric calcium imaging techniques. In aim-2, we will characterize the effects of TRPM3 dysfunction and deficiency on lens cation status, water content, and calcium influx properties using a combination of atomic absorption spectrometry and calcium imaging techniques. In aim-3, we will characterize the effects of TRPM3 dysfunction and deficiency on lens morphogenesis and gene expression using differential immuno-fluorescence microscopy and RNA deep-sequencing techniques, respectively. Results from these studies will provide new insights regarding the role of TRP-channels in lens growth, cation homeostasis, calcium dynamics, and pathophysiology within the context of pediatric cataract – a clinically important risk factor for lifelong visual impairment. More broadly, these studies will contribute to an understanding of TRP-channel function and dysfunction in other eye tissues (e.g., iris, ciliary body) and diseases (e.g., glaucoma, anterior eye defects), respectively.

项目概要/摘要非选择性阳离子通道的瞬时受体电位 (TRP) 超家族在多种疾病中发挥着关键作用生理过程包括对光、温度、压力和疼痛的感知。研究中，我们发现褪黑素相关 TRP 通道 (TRPM3) 成员的突变是一种遗传性儿童白内障的基础，与青光眼和眼前节有不同的相关性在本提案中，我们将研究 TRPM3 在晶状体发育和白内障病理学中的作用。通过使用基因编辑的小鼠和细胞系来解决三个特定目标，我们将描述镜头的特征。通过 RNA 深度测序和钙通道特性研究 TRP 通道转录物剪接变体在目标 2 中，我们将使用荧光钙成像技术来表征晶状体 TRPM3 亚型。 TRPM3 功能障碍和缺乏对晶状体阳离子状态、含水量和钙流入特性的影响在目标 3 中，我们将结合使用原子吸收光谱法和钙成像技术。描述 TRPM3 功能障碍和缺陷对晶状体形态发生和基因表达的影响分别使用差异免疫荧光显微镜和RNA深度测序技术。这些研究的结果将为 TRP 通道在晶状体生长、阳离子小儿白内障背景下的稳态、钙动态和病理生理学——临床研究更广泛地说，这些研究将有助于发现终生视力障碍的重要危险因素。了解其他眼组织（例如虹膜、睫状体）中的 TRP 通道功能和功能障碍，以及疾病（例如青光眼、前眼缺陷）。