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

• 批准号: 9889149
• 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
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-07 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9889149
• 关键词: 3D Print; Adopted; Adult; Affect; African; African American; Alteplase; Animal Model; Animals; Apolipoproteins; Apoptotic; Biological Models; 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; Pharmaceutical Preparations; Photosensitivity; Photosensitization; 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

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.

项目总结/摘要 中风是美国死亡的主要原因之一，也是成人残疾的主要原因， 社会负担。由脑动脉血栓栓塞性闭塞引起的缺血性中风， 大多数中风发生率，其余为血管破裂的出血性中风。在美国， 非裔美国人和白种人之间的中风发病率存在明显差异， （约240%）黑人。尽管对治疗进行了深入研究，但重组组织型纤溶酶原激活剂（rtPA） 仍然是FDA批准的唯一有效性有限的中风后药物。在这个可悲的不足背后， 中风治疗的缺乏在于难以产生足够大数量的中风诱导的神经元。 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导缺血性中风的选择方法。 光血栓形成是一种迅速采用的新方法，以诱导啮齿动物缺血性中风， 具有高度可重复的梗死面积和位置以及最小的死亡率等优点。在这个过程中，焦点 在暴露的颅骨上照射规定波长的光， （例如：孟加拉玫瑰红）在血液中，导致内皮细胞损伤和局部血小板聚集， 与受影响血管的堵塞有关然而，即使在这种情况下， 必须利用不容易按比例放大的数据来创建笔划模型。最近，光血栓性缺血性卒中 在成年斑马鱼脑中也通过在手动注射 Rose-Bengal，从而为中风研究提供了额外的脊椎动物模型系统。在这方面，委员会认为， 一种令人兴奋的新的遗传方法，它消除了人工注射光敏化学物质， 最近创建并测试成功，以选择性诱导光照射在成人细胞凋亡 斑马鱼心脏 在这里，我们提出了一个容易扩展的，光遗传学诱导中风系统的创建，使用 转基因斑马鱼，行为测试系统的开发，以确定新的治疗化学品， 并测试GWAS（全基因组关联研究）确定的人类APOL 1中风风险变体 基因，也被称为非洲血统人群中突出的肾脏疾病风险因素。