A scalable optogenetic system of focal stroke induction in zebrafish for testing stroke disparity genes

用于测试中风差异基因的斑马鱼局灶性中风诱导的可扩展光遗传学系统

基本信息

批准号：
10359724
负责人：
Ju-Ahng Lee
金额：
$ 11.1万
依托单位：
NORTH CAROLINA CENTRAL 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：
2019-03-07 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10359724
关键词：
3D Print Adopted Adult Affect African African American population African ancestry Alteplase Animal Model Animals Apolipoproteins Apoptotic Biological Models Black Populations Blood Circulation Blood Vessels Brain Brain hemorrhage Caucasians Cause of Death Cell Death Cerebrum Chemicals Chimeric Proteins Chronic Kidney Failure Data Drug Screening Effectiveness Embryo Endothelial Cells Endothelium FDA approved Frequencies Genes Genetic Heart Hemorrhage Human Incidence Individual Infarction Injections Injury Ischemic Stroke Kidney Diseases Light Lighting Location Manuals Measures Medicine Methods Middle Cerebral Artery Occlusion Modeling Mus Nature Operative Surgical Procedures Pathology Persons Pharmaceutical Preparations Photosensitivity Photosensitizing Agents Platelet aggregation Predisposition Procedures Reproducibility Research Rest Risk Risk Factors Rodent Rose Bengal Rupture Severities Stroke System Testing Therapeutic Transgenic Organisms Variant Vertebrates Zebrafish behavior test cerebral artery cranium design disability genetic approach genetic variant genome wide association study mortality novel novel strategies novel therapeutics optogenetics post stroke risk variant scale up stroke incidence stroke model stroke risk zebrafish development

项目摘要

PROJECT SUMMARY/ABSTRACT Stroke is one of the leading causes of death in the U.S. and also a leading cause of adult disability with severe societal burdens. Ischemic strokes, caused by the thromboembolic occlusion of cerebral arteries, constitute the majority of stroke incidences and the rest are hemorrhagic strokes with ruptured blood vessels. In the U.S., stroke incidence shows clear disparity between African Americans and Caucasians with much higher frequency (~240%) in blacks. Despite intensive search for treatment, recombinant tissue plasminogen activator (rtPA) remains the only FDA-approved post-stroke medicine with limited effectiveness. Behind this woefully inadequate dearth of stroke therapeutics lies the difficulties in generating a sufficiently large number of stroke-induced animals for effective drug screening, as a highly labor-intensive surgical procedure (middle cerebral artery occlusion) is still the method of choice to induce ischemic strokes in model animals. Photothrombosis is one of rapidly adopted new methods to induce ischemic strokes in rodents, with a number of advantages such as highly reproducible infarct size and location with minimal mortality. In this procedure, focal illumination of defined wavelength light on the exposed skull activates an IV-injected photosensitive chemical (eg. Rose-Bengal) in the bloodstream, causing injuries in endothelial cells and local platelet aggregation, leading to the clogging of the affected blood vessel. However, even in this case labor-intensive procedures that cannot not easily be scaled up must be utilized to create the stroke model. Recently, photothrombic ischemic stroke has also been successfully induced in the adult zebrafish brain by focal illumination following a manual injection of Rose-Bengal, thus providing an additional vertebrate animal model system for stroke research. In this regard, an exciting novel genetic approach, which eliminates the manual injection of photosensitive chemicals, was recently created and tested successfully to selectively induce apoptotic cell death by light illumination in the adult zebrafish heart. Here we propose a creation of a readily scalable, optogenetically induced stroke system using transgenic zebrafish, development of a behavioral test system to identify novel therapeutic chemicals, and to test GWAS (genome wide association study)-identified stroke risk variants of the human APOL1 gene, also known as prominent kidney disorder risk factors in people of African ancestry.

项目概要/摘要中风是美国的主要原因之一，也是导致成人严重残疾的主要原因社会负担。缺血性中风是由脑动脉血栓栓塞引起的大多数中风发生率和其余的是血管破裂的出血性中风。在美国，非裔美国人和白人之间的中风发病率存在明显差异，且发生率更高 (~240%) 为黑人。尽管积极寻找治疗方法，重组组织纤溶酶原激活剂（rtPA）仍然是 FDA 批准的唯一一种效果有限的中风后药物。这背后的不足之处中风治疗方法的缺乏在于难以产生足够大量的中风诱发的患者动物进行有效的药物筛选，作为一种高度劳动密集型的外科手术（大脑中动脉闭塞）仍然是在模型动物中诱发缺血性中风的首选方法。光血栓形成是快速采用的诱导啮齿动物缺血性中风的新方法之一，有许多具有高度可重复性的梗塞面积和位置以及最低死亡率等优点。在此过程中，焦点用规定波长的光照射暴露的头骨会激活静脉注射的光敏化学物质（如孟加拉玫瑰）进入血液，导致内皮细胞损伤和局部血小板聚集，导致受影响血管的堵塞。然而，即使在这种情况下，劳动密集型程序也无法必须利用不易按比例放大的来创建中风模型。近年来，光血栓性缺血性脑卒中手动注射后通过聚焦照明也成功地在成年斑马鱼大脑中诱导 Rose-Bengal，从而为中风研究提供了额外的脊椎动物模型系统。对此，一种令人兴奋的新型遗传方法，消除了手动注射光敏化学物质的过程最近创建并成功测试，可通过光照射选择性诱导成人细胞凋亡斑马鱼的心脏。在这里，我们建议使用以下方法创建一个易于扩展的光遗传学诱导中风系统转基因斑马鱼，开发行为测试系统来识别新型治疗化学品，并测试 GWAS（全基因组关联研究）确定的人类 APOL1 的中风风险变异基因，也被称为非洲血统人群中显着的肾脏疾病危险因素。