Neurovascular Unit on a Chip: Regional Chemical Communication, Drug and Toxin Responses

描述（由申请人提供）：全身血流和大脑之间的化学信号的物理或药理破坏损害大脑的正常功能和反应性。通过细胞因子、营养素、生长因子、激素、脂质、神经递质、药物及其代谢物的失调的长距离化学信号传导也很重要，但这些化学信号难以量化，并且通常在分离的情况下研究细胞。血脑屏障（BBB）动态地控制大脑和身体之间的交换，但这不能在完整的人脑中直接研究或通过动物模型充分代表。大多数现有的体外BBB模型不包括具有其他BBB元件的神经元和神经胶质，并且不能充分预测药物功效和毒性。这项研究将开发一个体外，三维，多隔室，中枢神经系统（CNS）神经血管单位（NVU）和脑脊液（CSF）隔室的器官型模型，两者都耦合到一个现实的血液替代供应系统，也包括循环免疫细胞。原代和干细胞衍生的人类细胞将与各种试剂相互作用，以产生跨越BBB和脑区域之间的关键化学通信，从而提供忠实地再现人类BBB、CNS和CSF的性质的紧凑装置。所提出的体外BBB/CNS/CSF模型将具有小体积，需要有限数量的人类细胞，可以重建不同脑区域之间的相互作用，并且将真实的时间耦合到先进的电化学和质谱仪器。这种变革性的技术平台将复制大脑中的化学通讯、分子运输和炎症，并将实现有针对性的和临床相关的营养和药理干预或预防。该平台将用于检查BBB在调节化学体-脑相互作用中的作用，表征大脑中的神经胶质细胞和神经细胞相互作用，并评估各种药物，化学品，感染剂和外源性物质对各种大脑区域的影响。该模型的临床实用性依赖于其以下能力：1）通过选择神经元、内皮细胞、星形胶质细胞、其他神经胶质细胞、周细胞和全身性白细胞的特定组合来重建独特区域，2）使用源自具有已知病理的患者的细胞和流体来评估药物治疗和来自慢性疾病如肥胖和急性损伤如中风的生理应激，3）揭示药物发现过程中的潜在副作用以及临床试验中使用的那些副作用，例如脑内皮细胞对批准药物的毒性转化，4）检测在BBB处会聚的大量化学信号之间的新颖且无偏的相关性，以及5）联合收割机微流体装置，最先进的细胞培养和器官型人脑细胞制备、分析仪器、生物信息学、控制理论和神经科学药物发现。综合方法将提供广泛适用的技术，并揭示大脑如何接收，修改和受药物，神经营养剂和疾病影响的新机制和区域特异性见解。

DESCRIPTION (provided by applicant): Physical or pharmacological disruption of chemical signals between the systemic blood flow and the brain im- pairs normal functioning and responsiveness of the brain. Long-range chemical signaling through dysregulation of cytokines, nutrients, growth factors, hormones, lipids, neurotransmitters, drugs and their metabolites is also important, but these chemical signals are difficult to quantify and cells are usually studied n isolation. The blood-brain barrier (BBB) dynamically controls exchange between the brain and body, but this cannot be studied directly in the intact human brain or adequately represented by animal models. Most existing in vitro BBB models do not include neurons and glia with other BBB elements and cannot adequately predict drug efficacy and toxicity. This research will develop an in vitro, three-dimensional, multi-compartment, organotypic model of a central nervous system (CNS) neurovascular unit (NVU) and cerebral spinal fluid (CSF) compartment, both coupled to a realistic blood-surrogate supply system that also incorporates circulating immune cells. Primary and stem-cell-derived human cells will interact with a variety of agents to produce critical chemical communications across the BBB and between brain regions, providing a compact device that faithfully reproduces the properties of the human BBB, the CNS, and the CSF. The proposed in vitro BBB/CNS/CSF model will have a small volume, requires a limited number of human cells, can recreate interactions between different brain regions, and will be coupled in real time to advanced electrochemical and mass spectrometry instruments. This transformative technological platform will replicate chemical communication, molecular trafficking, and inflammation in the brain, and will enable targeted and clinically relevant nutritional and pharmacologic interventions or prevention. This platform will be used to examine the role of the BBB in modulating chemical body-brain interactions, characterize glial and neural cell interactions in the brain, and assess the effect of a wide range of drugs, chemicals, infectious agents and xenobiotics on various brain regions. The model's clinical utility rests on its ability to 1) recreate unique regions by selecting specific combinations of neurons, endothelial cells, astrocytes, other neuroglia, pericytes and systemic leukocytes, 2) use cells and fluids derived from patients with known pathologies to assess drug treatments and physiological stress from chronic diseases such as obesity and acute injury such as stroke, 3) uncover potential adverse effects during drug discovery as well as those that are being used in clinical trials, such as toxic transformation of approved drugs by brain endothelial cells, 4) detet novel and unbiased correlations between large numbers of chemical signals which converge at the BBB, and 5) combine microfluidic devices, state-of-the-art cell culture and organotypic human brain-cell preparations, analytical instruments, bioinformatics, control theory, and neuroscience drug discovery. An integrated approach will provide technologies of widespread applicability and reveal new mechanistic and region-specific insights into how the brain receives, modifies, and is affected by drugs, neurotropic agents and disease.