Development of a 3D neurovascular unit for in vitro modeling of subarachnoid hemorrhage and screening therapies

项目摘要 在这个MOSAIC K99/R 00独立之路应用程序中，Brian O 'Grady博士提出了以下培训： 蛛网膜下腔出血（SAH）模型和治疗方法的开发， 他在仿生水凝胶中开发离体脑组织的小动脉特异性生长方面的专业知识， 3D打印微流体制造。培训计划与科学研究相结合，将开发和应用 用于模拟蛛网膜下腔出血中风事件并用作筛选的新型微流体装置 双靶向纳米颗粒作为SAH引起的损伤和延迟的潜在治疗的平台 脑缺血奥格雷迪博士的主要目标是成为一名独立的研究人员， 创建脑血管系统的仿生体外模型，并开发新的治疗方法， 神经系统疾病所描述的严格培训和血管生物学方面杰出的导师团队 (Dr. Lippmann）、神经疾病病理学（Jefferson博士）以及纳米颗粒开发和治疗学 (Dr.杜瓦尔）将确保他成功过渡到独立。通过他的训练计划，奥格雷迪博士将 获得1）血脑屏障生理学和神经血管单位的更深入的知识; 2）经验 合成和表征纳米颗粒; 3）对SAH和神经系统疾病建模的知识， 体外;和4）运行一个成功的跨学科和合作研究实验室的战略。 SAH被定义为以脑动脉瘤破裂为初始事件的脑血管疾病， 占所有中风类型的5%尽管这一比例很小，但SAH占所有中风相关疾病的三分之一。 在65岁之前失去的潜在寿命。虽然神经重症监护管理的新时代 为了改善SAH的预后，继发性后果导致迟发性脑缺血（DCI）。DCI具有 不同程度的患者功能结果，并且没有已知的干预措施来改善生活质量。这 缺乏有效的治疗方法在很大程度上归因于将脑靶向药物从 动物到人类最近，有一个全球性的努力，以产生组织工程，在体外模型 系统，可以代表复杂的血管解剖结构和神经血管单位的微环境。博士 O 'Grady的初步工作表明，一种新型仿生水凝胶支持诱导多能干细胞 细胞衍生的神经、壁和神经胶质细胞，并诱导离体人脑的小动脉特异性生长 脉管系统这种新的脉管系统由解剖学上正确的同心分层结构组成， 以前无法获得。当由微流体装置支撑时，小动脉扩张并且可以是管腔- 灌注和光消融。根据他的初步数据，奥格雷迪博士假设， 中风样事件的神经血管微环境可以通过这种新的体外系统准确地建模。 除了开发一种新的SAH体外模型外，本项目还将测试和验证神经保护剂 在人体外模型中，SAH和DCI的双靶向治疗剂的功效。

Project Summary In this MOSAIC K99/R00 Pathway to Independence application, Dr. Brian O’Grady proposes training in models of subarachnoid hemorrhage (SAH) and development of therapeutics that will strategically compliment his expertise in the development of arteriole-specific growth of ex vivo brain tissue in a biomimetic hydrogel and 3D printed microfluidic fabrication. The training plan is paired with scientific studies that will develop and apply a novel microfluidic device for modeling subarachnoid hemorrhage stroke events and for use as a screening platform for a dual-targeted nanoparticle as a potential therapeutic for the damage caused by SAH and delayed cerebral ischemia. Dr. O’Grady’s primary goal is to become an independent researcher focused on creating biomimetic in vitro models of the brain vasculature and developing novel therapeutics for neurological diseases. The rigorous training described and the outstanding team of mentors in vascular biology (Dr. Lippmann), neurological disease pathology (Dr. Jefferson), and nanoparticle development and therapeutics (Dr. Duvall) will ensure his success in transitioning to independence. Through his training plan, Dr. O’Grady will gain 1) deeper knowledge of blood-brain barrier physiology and the neurovascular unit; 2) experience synthesizing and characterizing nanoparticles; 3) knowledge of modeling SAH and neurological disorders in vitro; and 4) strategies for running a successful interdisciplinary and collaborative research lab. SAH is defined as a cerebrovascular disease with the initial event of a ruptured brain aneurysm and accounts for 5% of all types of strokes. Despite this small percentage, SAH accounts for one third of all stroke-related years of potential life lost before the age of 65. While a new era of neurocritical care management has contributed to improved outcomes for SAH, the secondary consequences result in delayed cerebral ischemia (DCI). DCI has varying degrees of patient functional outcome and has no known interventions to improve quality of life. This lack of effective treatments is largely attributed to the high failure rate of translating brain-targeting drugs from animals to humans. Recently, there has been a global effort to produce a tissue engineered, in vitro model system that can represent the complex vascular anatomy and microenvironment of the neurovascular unit. Dr. O’Grady’s preliminary work demonstrates that a novel biomimetic hydrogel supports induced pluripotent stem cell-derived neural, mural, and glial cells and induces arteriole-specific growth of ex vivo human brain vasculature. This new vasculature consists of anatomically correct, concentric layered structures that were previously unobtainable. When supported by a microfluidic device, the arterioles anastomose and can be lumen- perfused and photoablated. Based on his preliminary data, Dr. O’Grady hypothesizes that the dynamic neurovascular microenvironment of a stroke-like event can be accurately modeled by this new in vitro system. In addition to developing a new in vitro model of SAH, this project will test and validate the neural protective efficacy of a dual-targeted therapeutic for SAH and DCI in the human in vitro model.