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: 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
• 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

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.

项目摘要/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。由脑动脉的血栓栓塞阻塞引起的缺血性中风构成 大多数中风发生率和其余的都是出血性中风，血管破裂。在美国， 中风发生率显示出频率更高的非裔美国人和高加索人之间的明显差异 （〜240％）黑人。尽管密集地寻找治疗，但重组组织纤溶酶原激活剂（RTPA） 仍然是唯一经FDA批准的中风后药物，其有效性有限。在这个悲惨的不足之后 中风治疗的缺乏在于产生足够数量的中风引起的困难 动物进行有效的药物筛查，作为高度劳动密集型手术程序（大脑中动脉 闭塞）仍然是诱导模型动物缺血性中风的首选方法。 光链栓塞是一种迅速采用的新方法，以诱导啮齿动物的缺血性中风，其中一个 优点，例如高度可重复的梗塞大小和位置，死亡率最小。在此过程中，焦点 裸露的颅骨上定义的波长光的照明激活IV注入的光敏化学物质 （例如，玫瑰孟加拉）在血液中，在内皮细胞和局部血小板聚集中造成损伤，领先 堵塞受影响的血管。但是，即使在这种情况下，劳动密集型程序也无法 不容易扩大规模来创建中风模型。最近，光疗缺血性中风具有 在手动注射后，通过局灶性照明成功地在成年斑马鱼大脑中诱导了 玫瑰孟加拉，因此为中风研究提供了额外的脊椎动物动物模型系统。在这方面， 一种令人兴奋的新型遗传方法，它消除了光敏化学物质的手动注射，是 最近创建并成功测试了成人的光照明诱导凋亡细胞死亡 斑马鱼的心。 在这里，我们提出了一个易于扩展，光遗传学诱导的中风系统的创建 转基因斑马鱼，开发行为测试系统以识别新型治疗化学物质， 并测试GWAS（基因组广泛的关联研究）人类apol1的识别中风风险变体 吉恩（Gene），也称为非洲血统人群中著名的肾脏疾病危险因素。