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

描述(申请人提供)：系统血流和大脑之间的化学信号的物理或药物干扰，损害大脑的正常功能和反应。通过细胞因子、营养物质、生长因子、激素、脂质、神经递质、药物及其代谢物的失调而产生的远程化学信号也很重要，但这些化学信号很难量化，而且通常是通过分离来研究细胞的。血脑屏障(BBB)动态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，也不能用动物模型充分代表。现有的大多数体外血脑屏障模型不包括神经元和神经胶质细胞与其他血脑屏障成分，不能充分预测药物的疗效和毒性。这项研究将开发一种体外、三维、多隔室、器官型的中枢神经系统(CNS)神经血管单位(NVU)和脑脊液(CSF)隔室模型，两者都耦合到一个现实的血液代理供应系统，该系统也结合了循环免疫细胞。原代和干细胞来源的人类细胞将与各种试剂相互作用，在血脑屏障和脑区域之间产生关键的化学通信，提供一种紧凑的设备，忠实地再现人类血脑屏障、中枢神经系统和脑脊液的特性。所提出的体外BBB/CNS/CSF模型体积小，需要有限的人体细胞，可以重建不同脑区之间的相互作用，并将实时耦合到先进的电化学和质谱仪。这一变革性的技术平台将复制大脑中的化学通信、分子运输和炎症，并将使有针对性和临床相关的营养和药物干预或预防成为可能。这一平台将被用来研究血脑屏障在调节身体-大脑化学相互作用中的作用，表征大脑中神经胶质细胞和神经细胞的相互作用，并评估一系列药物、化学品、感染剂和异物对大脑不同区域的影响。该模型的临床实用性取决于以下能力：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、周细胞和系统性白细胞的特定组合来重建独特的区域；2)使用来自已知病理的患者的细胞和液体来评估药物治疗和诸如肥胖和中风等急性损伤等慢性疾病的生理应激；3)在药物发现期间发现潜在的不良反应以及用于临床试验的潜在不良反应，例如大脑内皮细胞对批准的药物的毒性转化；4)检测汇聚在血脑屏障的大量化学信号之间的新的和无偏见的相关性；以及5)结合微流控设备，最先进的细胞培养和人脑细胞器官型制剂、分析仪器、生物信息学、控制理论和神经科学药物发现。一种综合的方法将提供广泛适用的技术，并揭示大脑如何接受、修改和受药物、神经促进剂和疾病影响的新的机械性和特定于区域的见解。

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.