Identifying targeting antibodies in an in vitro model of the human blood-brain barrier during ischemic stroke

在美国，中风是导致残疾和死亡的主要原因之一。造成这一现象的原因之一 缺乏FDA批准的慢性中风治疗方法。除组织纤溶酶原激活物(TPA)外， 没有其他已获批准的疗法具有改善疾病的效果；目前的疗法完全集中在 关于症状管理和康复。这在一定程度上是由于对 涉及的潜在机制，以及通过血脑传递治疗药物的困难 屏障(BBB)和进入大脑。这个项目的目的是通过 人诱导多能干细胞(HiPSC)BBB掺入模型的建立 HIPSC来源于脑微血管内皮样细胞(BMECs)、星形胶质细胞和神经元。使用3D 微血管平台缺血将使用剪切流、氧浓度、 和营养素的可用性。由此产生的细胞表型将使用活细胞显微镜进行研究。 测量荧光示踪剂分子的通透性，以及细胞的形态变化。 使用这个模型，以及一个简化的2D缺血模型，将对抗体库进行筛选 改进了对人类缺血脑的靶向和递送。最后，在屏幕上识别的抗体 将测试将已知的非脑穿透性神经保护性生长因子输送到 在体外模型和中风小鼠模型中都存在脑缺血。总而言之，这个项目将 建立人类在缺血性卒中期间血脑屏障的特异性模型，并识别能够 专门针对大脑的缺血区并提供治疗药物。

Stroke is one of the leading causes of disability and death in the US. One of the contributing factors to this is the lack of FDA approved treatments for chronic stroke. Aside from tissue plasminogen activator (tPA), there are no other approved therapeutics with disease modifying effects; current therapy is entirely focused on symptom management and rehabilitation. This is in part due to the lack of understanding of the underlying mechanisms involved, as well as the difficulty of delivering therapeutics through the blood-brain barrier (BBB) and into the brain. This project aims to address both of these difficulties through the development of a human induced pluripotent stem cell (hiPSC) derived model of the BBB incorporating hiPSC derived brain microvascular endothelial like cells (BMECs), astrocytes, and neurons. Using a 3D microvessel platform ischemia will be modeled using perturbations in shear flow, oxygen concentration, and nutrient availability. The resultant phenotype of the cells will be investigated using live cell microscopy to measure permeability of fluorescent tracer molecules, as well as morphological changes in the cells. Using this model, as well as a simplified 2D model of ischemia, a library of antibodies will be screened for improved targeting and delivery to the human ischemic brain. Finally, the antibodies identified in the screen will be tested for efficacy in delivering known non-brain penetrant neuroprotectant growth factors to the ischemic brain both in the in vitro model and in a mouse model of stroke. In summary, this project will establish a human specific model of the BBB during ischemic stroke and identify antibodies that are able to specifically target and deliver therapeutics to the ischemic region of the brain.