Development of a BBB model to study transendothelial cell migration

开发 BBB 模型来研究跨内皮细胞迁移

• 批准号: 8314680
• 负责人: DAMIR JANIGRO
• 金额: $ 56万
• 依托单位: FLOCEL, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8314680
• 关键词: Achievement; Anti-Inflammatory Agents; Anti-inflammatory; Arts; Biological; Blood - brain barrier anatomy; Blood Vessels; Blood capillaries; Brain; Brain Diseases; Cell Line; Characteristics; Clinical; Coculture Techniques; Contracts; Data; Development; Disease; Drug resistance; Economics; Endothelial Cells; Epilepsy; Experimental Designs; Extravasation; Fee-for-Service Plans; Fiber; Human; Immune response; In Vitro; Inflammatory; Knowledge; Leukocyte Trafficking; Leukocytes; Mechanics; Mediation; Modality; Modeling; Multiple Sclerosis; Neuroglia; Pathologic; Pattern; Performance; Pericytes; Permeability; Pharmaceutical Preparations; Pharmacological Treatment; Pharmacology; Phase; Physiological; Physiology; Process; Progress Reports; Property; Protocols documentation; Publishing; Recording of previous events; Reproduction; Research Personnel; Rest; Rodent; Role; Scanning Electron Microscopy; Side; Signal Transduction; Small Business Technology Transfer Research; Smooth Muscle Myocytes; Stimulus; Stretching; System; Test Result; Testing; Traction; base; capillary; cell motility; chemokine; clinically relevant; commercialization; design; drug development; drug testing; drug use screening; hemodynamics; improved; in vitro Model; insight; migration; monocyte; nervous system disorder; novel; pre-clinical; prototype; research and development; research study; response; scale up; shear stress; sound; success; therapeutic target; venule

DESCRIPTION (provided by applicant): Reproducing in vitro the physiological characteristics of brain vascular segments represents a critical issue. Adequate modeling of the cerebrovasculature could significantly help understand the mechanisms and improve the pharmacology of disease where a role for leukocytes migrating across the cerebrovasculature is demonstrated. Thanks to the previous support (Phase I) we were able to prototype and test drive a new dynamic in vitro model of the BBB (DIV- BBB) permissive for leukocyte extravasation. Our initial effort aimed at piercing hollow fibers in a reproducible manner; the results were recently published. We found that manually perforated hollow fibers allow leukocytes passage across the BBB in response to pro-inflammatory stimuli and hemodynamic changes. We were also able to produce a capillary-venule segment by varying the rheological parameters (e.g., changing the shear stress) of the system. In addition we recently developed a system to mechanically "stretch" the hollow fibers increasing the pore size up to the physiologically relevant size of ~5 ¿m. Controlled traction was applied to the end of the fibers and scanning electron microscopy showed enlarged pores within the stretched fiber. To further the commercial opportunity afforded by this new BBB model, we propose the following Phase II Specific Aims: To optimize the performance of a dynamic in vitro capillary-venules model of the brain cerebrovasculature permissive for leukocyte extravasation. To determine the pattern of leukocytes extravasation in control and diseased capillary-venules segments composed of fibers with different transmural permeability properties (from Aim 1). To compare the results obtained using these DIV- BBB models to other state-of-art in vitro BBB models. We will initially tailor the use of the DIV capillary-venules system to multiple sclerosis and epilepsy research and drug development. We have assembled a multi-disciplinary team of investigators and experts in the field of leukocyte migration across the cerebrovasculature. Additional clinically relevant venues are detailed in the Commercialization Plan. Given our preliminary results and the confirmatory progress report described in detail in this application, and given the fact that i the meantime development of new drugs has remained a major issue in the treatment of neurological diseases, we believe that the combination of a strong record of accomplishment and sound experimental design aimed at improving drug development are fundamental aspects of this Phase 2 proposal. PUBLIC HEALTH RELEVANCE: Leukocyte migration into the brain parenchyma is a hallmark of various neuro-inflammatory brain diseases. In order to develop therapeutics targeting brain inflammatory process it is imperative to reproduce in vitro the modality by which leukocytes cross the blood-brain barrier (BBB) to reach the brain parenchyma. We now plan to optimize the performance of a dynamic in vitro capillary-venules model of the brain cerebrovasculature permissive for leukocyte extravasation. We will also determine the pattern of leukocytes extravasation in control and diseased (multiple sclerosis and epilepsy) capillary-venules segments composed of fibers with different permeability properties.

描述（由申请人提供）：在体外复制脑血管段的生理特征是一个关键问题，脑血管系统的充分建模可以显着帮助理解疾病的机制并改善疾病的药理学，其中白细胞在脑血管系统中迁移的作用已得到证实。感谢之前的支持（第一阶段），我们能够制作原型并测试驱动 BBB (DIV- BBB) 许可的新动态体外模型我们最初的努力旨在以可重复的方式刺穿中空纤维；我们发现手动穿孔的中空纤维允许白细胞通过血脑屏障，以响应促炎刺激和血流动力学变化。通过改变系统的流变参数（例如，改变剪切应力）来产生毛细血管段。此外，我们最近开发了一种机械“拉伸”中空纤维的系统。孔径可达生理相关尺寸 ~5 ¿ m. 对纤维末端施加受控牵引力，扫描电子显微镜显示拉伸纤维内的孔隙扩大。为了进一步利用这种新的 BBB 模型提供的商业机会，我们提出了以下第二阶段的具体目标：允许白细胞外渗的脑脑血管系统的动态体外毛细血管微静脉模型以确定对照和患病的白细胞外渗模式。由具有不同透壁渗透性的纤维组成的毛细血管段（来自目标 1） 为了将使用这些 DIV-BBB 模型与其他最先进的体外 BBB 模型获得的结果进行比较，我们将首先调整 DIV 的使用。我们组建了一支由白细胞跨脑血管迁移领域的研究人员和专家组成的多学科团队。鉴于我们的初步结果和本申请中详细描述的验证性进展报告，并考虑到新药的开发仍然是神经系统疾病治疗中的一个主要问题，商业化计划中详细介绍了其他临床相关场所。 ，我们相信，良好的成就记录和旨在改善药物开发的合理实验设计的结合是第二阶段提案的基本方面。 公共健康相关性：白细胞迁移到脑实质是各种神经炎症性脑疾病的标志，为了开发针对脑炎症过程的治疗方法，必须在体外重现白细胞穿过血脑屏障（BBB）的方式。 ）以到达脑实质。我们现在计划优化允许白细胞的脑脑血管系统动态体外毛细血管模型的性能。我们还将确定由具有不同渗透特性的纤维组成的对照和患病（多发性硬化症和癫痫）毛细血管小静脉段中白细胞外渗的模式。