Side-by-side comparison of blood-brain barrier models

描述（由申请人提供）：血脑屏障（BBB）对于复杂问题至关重要，例如药物递送、涉及BBB功能障碍的慢性神经系统疾病的发病机制（例如，脑肿瘤、缺血、缺氧、脑水肿、多发性硬化和脑膜炎）以及与生物防御相关的问题。由于血脑屏障选择性地（通过特定的运输机制）排除大多数血液传播的物质和外源性物质进入大脑，保护它免受全身影响。不幸的是，生物防御系统的发展，以保护大脑从潜在的危险物质也可能有助于在治疗几个中枢神经系统疾病，如药物难治性癫痫或难治性脑肿瘤的现象称为多药耐药性（MDR）。制药公司每年花费数百万美元来开发绕过脑实质屏蔽的替代药物策略，并使用BBB的体内或体外模型研究新的治疗方法，其中许多最终不起作用。CNS药物设计不能完全和完全依赖于推定的神经治疗剂的物理化学性质，因为亲脂性本身是药物渗透到CNS的不良预测因子。这对于CNS药物、抗癫痫药、抗病毒药和抗癫痫药的3大家族尤其如此。在小动物（包括啮齿动物）中进行的研究不能直接外推至人体组织。该实验室和其他实验室的初步结果令人信服地证明，使用啮齿动物脑内皮细胞和一般非人类来源的内皮细胞系作为临床药理学模型是有缺陷的。我们建议：1）研究在动态条件下生长的内皮-胶质细胞共培养物中多药耐药表达的影响，并比较基于Transwell型技术的可比BBB模型中获得的渗透性值。2)确认当前钉仓设计，并对现有DIV-BBB进行改进，以解决当前设计问题。3)验证电子测量系统的设计。当前TEER设计的改进是能够在各种频率下测量BBB的阻抗。我们建议开发和验证一种改进的动态体外血脑屏障（DIV-BBB）模型，该模型再现了体内血脑屏障的功能特征，具有更高的可预测性，并且完全可扩展和可定制。因此，它将非常适合广泛的药理学和生理学研究，并将加速导致针对CNS疾病的新的和更有效的药物疗法的过程。

DESCRIPTION (provided by applicant): The blood-brain barrier (BBB) is crucial for complex issues such as drug delivery, pathogenesis of chronic neurological diseases involving BBB dysfunction (e.g., brain tumors, ischemia, hypoxia, brain edema, multiple sclerosis, and meningitis) and issues related to bio-defense. Since the BBB selectively (by specific transport mechanisms) excludes most blood-borne substances and xenobiotics from entering the brain, protecting it from systemic influences. Unfortunately, bio-defense systems that developed to protect the brain from potentially dangerous substances may also contribute to the phenomenon known as multiple drug resistance (MDR) during treatment of several CNS disorders, such as drug refractory epilepsy or intractable brain tumors. Every year millions of dollars are spent by pharmaceutical companies to develop alternative pharmaceutical strategies that bypass the shielding of brain parenchyma and to study new therapeutic approaches using in vivo or in vitro models of the BBB, many of which end up not working. Rationale CNS drug design cannot entirely and exclusively rely upon the physical-chemical properties of putative neurotherapeutics, since lipophilicity alone is a poor predictor for drug penetration into the CNS. This is particularly true for 3 large families of CNS drugs, antineoplastics, antivirals and antiepileptics. Studies performed in small animals including rodents cannot be directly extrapolated to human tissue. Preliminary results from this and other laboratories have convincingly demonstrated that use of rodent brain endothelial cells and in general endothelial cell lines from non human sources as models of clinical pharmacology are flawed. We propose to: 1) To study the effects of multiple drug resistance expression in endothelial-glial co-culture grown under dynamic conditions and to compare permeability values obtained in a comparable BBB model based on Transwell-type technology. 2) To validate the current design of the cartridge and implement improvements to the existing DIV-BBB to address current deign problems. 3) To validate the design of the electronic measurement system. The improvement in the current TEER design is the ability to measure the impedance of BBB at various frequencies. We propose to develop and validate an improved dynamic in vitro blood-brain barrier (DIV-BBB) model that reproduces the functional characteristics of the BBB in vivo, features higher predictability, and is fully scalable and customizable. As such, it will be perfectly suited for extensive pharmacological and physiological studies and will accelerate the process leading to new and more effective drug therapies aimed at CNS diseases.