Targeting the Blood-Brain Barrier in Neuroinflammation

针对神经炎症中的血脑屏障

• 批准号: 6559719
• 负责人: ULRICH BICKEL
• 金额: $ 26.52万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-01-01 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6559719
• 关键词: antiinflammatory agents; biotherapeutic agent; blood brain barrier; cell line; cerebrovascular system; drug delivery systems; drug design /synthesis /production; experimental allergic encephalomyelitis; inflammation; laboratory mouse; monoclonal antibody; nitric oxide synthase; nuclear factor kappa beta; oligonucleotides; pharmacokinetics; polyethylene glycols; polyethylenes; prostaglandin endoperoxide synthase; transferrin receptor; vascular endothelium

DESCRIPTION: [Applicant's Abstract] This proposal explores a novel use of transferrin receptor mediated drug delivery. An oligonucleotide (ODNbased drug will be targeted to brain microvascular endothelial cells, with the goal to inhibit inflammatory responses in these cells. The approach could have broad applicability for diseases affecting the blood-brain barrier and the central nervous system. Here, we focus on experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis, to demonstrate the feasibility of this vascular targeting strategy. Crucial events in the pathogenesis of that disease affect function and integrity of the BBB. The endothelial cells forming the BBB express adhesion molecules affecting lymphocyte transmigration, and enzymes such as inducible NO-synthase and cyclooxygenase-2 contributing to inflammation. Expression of these proteins is under control of the transcription factor NF-kappaB. Therefore, preventing NF-kappaB activation could be a promising therapeutic strategy. An elegant way to inhibit a transcription factor is the cellular delivery of decoy ODNs, short double stranded ODNs containing the consensus binding sequence of that factor. Unfortunately, decoy ODNs face similar problems as related antisense approaches, namely poor permeability through cell membranes. The receptor-mediated endocytosis provided by the transferrin receptor system can overcome that obstacle. Here, the 8D3 antibody specific for the mouse transferring receptor will serve as the vector for delivery of a NF-kappaB decoy ODN. The polyanionic ODN will be bound in a complex by the cationic polymer polyethylenimine, and then coupled to the 8D3 antibody via an avidin-biotin bridge. Polyethylenimine is an attractive carrier for DNA due to its capacity to mediate "endosomal escape", thus enabling the ODNs to reach their cytosolic or nuclear sites of action. The Specific Aims of the project comprise (i) the determination of the cellular uptake and pharmacological activity of the ODN delivery system in an in vitro model with a murine brain endothelial cell line; (ii) determining the in vivo pharmacokinetics of the delivery system in normal mice; and (iii) evaluating in vivo pharmacological effects of vector-mediated ODN delivery in the mouse EAE-model.

描述：[申请人的摘要]该提案探索了转铁蛋白受体介导的药物递送的新用途。一种基于寡核苷酸（ODN）的药物将靶向脑微血管内皮细胞，目的是抑制这些细胞的炎症反应。该方法可能对影响血脑屏障和中枢神经系统的疾病具有广泛的适用性。在这里，我们重点关注实验性自身免疫性脑脊髓炎（EAE）是一种多发性硬化症动物模型，旨在证明这种血管靶向策略的可行性。该疾病发病机制中的关键事件会影响血管的功能和完整性。 BBB。形成 BBB 的内皮细胞表达影响淋巴细胞迁移的粘附分子，以及导致炎症的诱导型 NO 合酶和环氧合酶 2 等酶，这些蛋白的表达受到转录因子 NF-κB 的控制。 -kappaB 激活可能是抑制转录因子的一种优雅方法，即诱饵 ODN（包含共有结合的短双链 ODN）的细胞递送。该因素的顺序。不幸的是，诱饵 ODN 面临与相关反义方法类似的问题，即细胞膜渗透性差。转铁蛋白受体系统提供的受体介导的内吞作用可以克服这一障碍。在此，针对小鼠转移受体的 8D3 抗体将作为载体用于递送 NF-kappaB 诱饵 ODN。聚阴离子 ODN 将通过阳离子聚合物聚乙烯亚胺结合成复合物，然后通过亲和素-生物素桥偶联至 8D3 抗体。聚乙烯亚胺是一种有吸引力的 DNA 载体，因为它能够介导“内体逃逸”，从而使 ODN 能够到达其细胞质或细胞核的作用位点。该项目的具体目标包括 (i) 在小鼠脑内皮细胞系体外模型中测定 ODN 递送系统的细胞摄取和药理活性； (ii) 确定正常小鼠体内递送系统的药代动力学； (iii) 评估载体介导的 ODN 递送在小鼠 EAE 模型中的体内药理作用。