A Vascularized Blood-Brain Barrier Model for In Vitro Testing of Drug and Immunotherapy Delivery

用于药物和免疫治疗递送体外测试的血管化血脑屏障模型

• 批准号: 10699597
• 负责人: G. WESLEY HATFIELD
• 金额: $ 29.5万
• 依托单位: ARACARI BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10699597
• 关键词: 3-Dimensional; Arteries; Astrocytes; Biological Sciences; Bispecific Antibodies; Blood; Blood - brain barrier anatomy; Blood Vessels; Bone Marrow; Brain; Brain Diseases; Brain Neoplasms; Carboplatin; Cells; Cisplatin; Client; Collaborations; Data; Devices; Dextrans; Disease Progression; Doxorubicin; Drug Targeting; Endothelial Cells; Environment; Equilibrium; Extracellular Matrix; Extravasation; Foot Process; Glioma; Goals; Homeostasis; Human; Immune; Immune system; Immunooncology; Immunotherapeutic agent; Immunotherapy; Left; Legal patent; Leukocytes; Lymphocyte; Marketing; Membrane; Microfluidic Microchips; Microfluidics; Modeling; Mus; Natural Killer Cells; Nutrient; Organ; Patients; Penetrance; Perfusion; Pericytes; Permeability; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Physiological; Physiology; Pinocytosis; Plasma; Polymers; Property; Proteins; Research; Small Business Innovation Research Grant; Speed; Surface; System; T-Lymphocyte; Technology; Tight Junctions; Up-Regulation; Vascularization; Veins; Work; bevacizumab; chimeric antigen receptor T cells; commercialization; design; drug discovery; drug testing; fluorescence imaging; in vitro Model; in vitro testing; in vivo; in vivo Model; monocyte; monolayer; mouse model; neoplastic cell; neurovascular unit; response; rhodamine dextran; small molecule; success; tool; tumor; γδ T cells

PROJECT SUMMARY The brain is a highly-specialized, finely-tuned organ that depends for its function on maintaining homeostasis within narrowly-defined limits. This is achieved in large part by the Blood-Brain Barrier (BBB), which tightly regulates what can and can’t get into the brain. Functionally, the BBB is the combined work of endothelial cells (EC), pericytes and astrocytes, the latter two of which act on the EC to maintain an extensive network of tight junctions, promote the expression of specialized transporters, and limit the rate of transcellular pinocytosis. This lack of free transfer between blood and brain for non-lipophilic species severely limits the entry of many, otherwise useful, small molecule drugs. For example, cisplatin (<3% Brain/Plasma ratio), is effective in non-brain tumors, but has poor penetrance and low efficacy against brain tumors such as glioma. Similarly, delivery of immunotherapeutics such as bevacizumab (0.2%) and bi-specific antibodies is also limited, and much is still to be understood regarding the delivery of immune cells to the brain, including CAR-Ts, NK cells, γδ T cells and the patient’s own monocytes and lymphocytes. Finally, there are significant differences between mouse and human physiology including in the immune system. Many drug developers are therefore turning to human cell- based systems rather than mouse models. The primary goal of this Phase I SBIR application is to demonstrate the feasibility of Aracari Biosciences’ proprietary technology to model appropriate drug and immunotherapeutic delivery across the human BBB. Aracari’s patented core technology is a perfusable human vascular network within a microfluidic device that fits in a convenient 96-well plate format. This technology has been successfully developed into commercialized products including the vascularized micro-organ (VMOTM) and the vascularized micro-tumor (VMTTM). Importantly, leukocytes can be perfused through the vessels and on stimulation they will adhere and extravasate as they do in vivo. We have also developed a vascularized micro-brain (VMB), which is the focus of this application. This incorporates a perfused neurovascular unit (NVU) comprised of human BBB EC, pericytes and astrocytes, all embedded in a brain-mimicking extracellular matrix. The vessels show upregulation of BBB transporters and junctional proteins, and greatly-reduced permeability compared to non- NVU vessels. Tumor cells (glioma) can be added to model brain tumors (VMB-T) and the potential compromise of the BBB. This model will provide unprecedented opportunities to study drug responses of glioma and other brain tumors in a more natural environment. Our preliminary data show the feasibility of our goals which are: Specific Aim 1: Characterize permeability of a small panel of brain-targeted drugs in the VMB and VMB-T Specific Aim 2: Characterize delivery of a small panel of immunotherapeutic drugs in the VMB and VMB-T Specific Aim 3: Characterize delivery of immune cells in the VMB and VMB-T Completion of these aims will provide us with a unique tool we can offer to pharmaceutical companies for studies on BBB properties, and delivery of neuroactive drugs and immuno-oncology drugs and cells.

项目摘要 大脑是一个高度专业化的、精细调节的器官，其功能依赖于维持体内平衡 在狭窄定义的范围内。这在很大程度上是通过血脑屏障（BBB）实现的， 控制着什么能进入大脑什么不能进入在功能上，血脑屏障是内皮细胞的联合工作， (EC)，周细胞和星形胶质细胞，后两者作用于EC以维持广泛的紧密网络。 连接，促进专门的转运蛋白的表达，并限制跨细胞胞饮的速率。这 非亲脂性物质在血液和脑之间缺乏自由转移严重限制了许多， 其它有用的小分子药物。例如，顺铂（<3%脑/血浆比率）在非脑肿瘤中有效。 肿瘤，但对脑肿瘤如神经胶质瘤的抑制率差且功效低。同样，交付 免疫治疗如贝伐单抗（0.2%）和双特异性抗体也是有限的，还有很多工作要做。 术语“免疫细胞”应理解为关于向脑递送免疫细胞，包括CAR-T、NK细胞、γδ T细胞和 患者自身的单核细胞和淋巴细胞。最后，小鼠和 包括免疫系统在内的人体生理学。因此，许多药物开发人员转向人类细胞- 而不是鼠标模型。第一阶段SBIR应用的主要目标是证明 Aracari Biosciences专有技术模拟适当药物和免疫系统的可行性 通过人血脑屏障递送。Aracari的专利核心技术是一种可灌注的人体血管网络 在适合于方便的96孔板形式的微流体装置内。该技术已成功 开发成商业化产品，包括血管化微器官（VMO TM）和血管化微器官（VMO TM）。 微肿瘤（VMTTM）。重要的是，白细胞可以通过血管灌注，并且在刺激下， 就像它们在体内一样粘附和渗出。我们还开发了一种血管化的微型大脑（VMB）， 这个应用程序的重点。这包括由人血脑屏障组成的灌注神经血管单位（NVU） EC，周细胞和星形胶质细胞，都嵌入在模拟脑的细胞外基质中。这些血管显示 BBB转运蛋白和连接蛋白的上调，以及与非 NVU船只。肿瘤细胞（胶质瘤）可以添加到模型脑肿瘤（VMB-T）中， 的BBB。该模型将为研究胶质瘤和其他肿瘤的药物反应提供前所未有的机会。 在更自然的环境中生长脑瘤我们的初步数据显示了我们目标的可行性，这些目标是： 具体目标1：表征一小组脑靶向药物在VMB和VMB-T中的渗透性 具体目标2：表征VMB和VMB-T中一小组免疫抑制药物的递送 具体目标3：表征免疫细胞在VMB和VMB-T中的递送 这些目标的完成将为我们提供一个独特的工具，我们可以提供给制药公司的研究 对血脑屏障性质的影响，以及神经活性药物和免疫肿瘤药物和细胞的递送。