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

项目摘要 在这个马赛克K99/R00独立之路应用程序中，Brian O‘Grady博士建议培训 蛛网膜下腔出血(SAH)模型和治疗方法的发展将在战略上相互补充 他在开发体外脑组织微动脉特异性生长仿生水凝胶方面的专业知识和 3D打印微流控制造。培训计划与科学研究相结合，这些研究将制定并应用 用于蛛网膜下腔出血卒中事件模拟和筛查的新型微流控装置 双靶向纳米颗粒平台作为治疗SAH和延迟损伤的潜在药物 脑缺血。奥格雷迪博士的主要目标是成为一名专注于 建立脑血管的体外仿生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优秀的血管生物学导师团队 (李普曼博士)，神经疾病病理学(杰斐逊博士)，纳米颗粒开发和治疗学 (杜瓦尔博士)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)更深入的血脑屏障生理学和神经血管单位的知识；2)经验 纳米粒子的合成和表征；3)对蛛网膜下腔出血和神经系统疾病的建模知识。 4)成功运作跨学科和协作研究实验室的策略。 蛛网膜下腔出血被定义为以脑动脉瘤破裂为初始事件的脑血管疾病 适用于所有类型中风的5%。尽管这一比例很小，但SAH占所有与中风相关的疾病的三分之一 65岁之前失去的潜在寿命年数。虽然神经危重护理管理的新时代已经做出了贡献 为了改善SAH的预后，继发性后果导致迟发性脑缺血(DCI)。DCI有 不同程度的患者功能结局，并且没有已知的干预措施来改善生活质量。这 缺乏有效的治疗在很大程度上是由于将脑靶向药物从 从动物到人类。最近，全球正在努力生产一种组织工程化的体外模型 可以代表复杂的血管解剖和神经血管单位的微环境的系统。Dr。 O‘Grady的初步工作表明，一种新的仿生水凝胶支持诱导的多能干细胞 细胞来源的神经细胞、壁细胞和神经胶质细胞并诱导体外人脑小动脉特异性生长 脉管系统。这种新的血管系统由解剖学上正确的同心分层结构组成 以前得不到的。当由微流体装置支撑时，小动脉吻合并可管腔- 灌流和光消融。根据他的初步数据，奥格雷迪博士假设 这一新的体外系统可以准确地模拟中风样事件的神经血管微环境。 除了开发一种新的SAH体外模型外，该项目还将测试和验证神经保护 双靶点治疗SAH和DCI在人体体外模型中的疗效。

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.