Bioengineering of the blood-brain barrier permeability

血脑屏障通透性的生物工程

• 批准号: 7250938
• 负责人: SERGUEI V VINOGRADOV
• 金额: $ 19.34万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7250938
• 关键词: Animal Model; Back; Binding; Biomedical Engineering; Blood; Blood - brain barrier anatomy; Blood Circulation; Brain; Brain Neoplasms; Capillary Endothelial Cell; Carrier Proteins; Cell Line; Cells; Central Nervous System Viral Diseases; Characteristics; Cultured Cells; DNA delivery; Data; Development; Disease; Dose; Down-Regulation; Drug Efflux; Drug Transport; Encapsulated; Endothelial Cells; Endothelium; Epithelial Cells; Ethylene Glycols; Eukaryota; Eukaryotic Cell; Ganciclovir; Gene Expression; Genes; Genetic; Goals; Homing; In Vitro; Lead; Libraries; Luciferases; Membrane; Membrane Proteins; Messenger RNA; Methods; Molecular Weight; Multi-Drug Resistance; Mus; NanoGel; Neuraxis; P-Glycoprotein; P-Glycoproteins; Patients; Peptides; Permeability; Phage Display; Pharmaceutical Preparations; Phenotype; Plasmids; Polymers; Polynucleotides; Principal Investigator; Process; Property; RNA Interference; Reagent; Resistance; Route; Small Interfering RNA; Structure; Structure of choroid plexus; Surface; System; Therapeutic; Therapeutic Agents; Thioguanine; Toxin; Transfection; Viral; Virus Diseases; Xenobiotics; base; blood cerebrospinal fluid barrier; chemical synthesis; chemotherapy; clinical application; crosslink; day; design; ethylene glycol; gene delivery system; gene therapy; in vivo; inhibitor/antagonist; lipophilicity; membrane activity; nanosized; non-viral gene delivery; non-viral gene therapy; nucleoside analog; particle; plasmid DNA; programs; research study; small hairpin RNA; stem; targeted delivery; tool; vector; viral gene delivery

DESCRIPTION (provided by applicant): Permeability of various drugs across the blood-brain barrier (BBB) is significantly dependent on the expression and functional activity of specific efflux transporters located in the membrane of brain capillary endothelial cells (BCEC). Selective transient downregulation of the transporters will lead to the application of more effective and less toxic doses of therapeutic drugs against brain tumors or viral infections in CNS. Previously, antisense inhibitors have been shown to temporarily arrest the synthesis of major multidrug resistance agent, membrane P-glycoprotein, and promote reversal of the resistant cell phenotype. The more effective RNA interference mechanism has been recently discovered for selective switching off expression of various genes. Short hairpin RNA (siRNA) could be introduced into target cells through a plasmid DNA precursor using methods of non-viral gene therapy. However, targeted delivery of the pDNA to the cells of the BBB requires a good systemic carrier and selective vectors that bind to the BCEC. As such a carrier, polymer crosslinked Nanogel particles modified with the brain-specific homing peptides (BSHP) have been chosen for tranfection of the BBB by shRNA-encoding plasmid DNA with an ultimate goal suppressing the specific membrane proteins, drug efflux transporters, in the BBB. Specific BSHPs to be attached to the surface of the Nanogel and target delivery to the BBB have been selected in vivo from a vast amount of peptides in the phage display library. Nanogel is non-toxic and highly effective as a transfection agent in many cell lines and evidently, one of the carriers with great potential for systemic administration. The vectorized RNA Interference-Producing system (RIP system) could be used for bioengineering of the BBB permeability for therapeutic agents whose brain accessibility was hampered by specific drug efflux transporters. The central hypothesis of the proposal is that transient suppression of selected drug efflux transporters in the BBB via systemic transfection of brain endothelium using targeted RNAi-producing systems can result in significant enhancement of drug transport to the brain during chemotherapy of the CNS-related diseases. Our Specific aim 1 is to develop the BCEC-targeted Nanogel carriers for systemic delivery of plasmid DNA to the BBB. Specific aim 2 is the enhance transfection efficacy of the BCEC-targeted Nanogel carriers in vitro and in vivo. Specific aim 3 is to suppress selected drug efflux transporters in the BBB in vivo and temporary increase drug transport into the brain. In this Aim brain transport of several representive nucleoside analogue drugs will be assessed in animal model following the transient downregulation of drug efflux transporters in the BBB by the Nanogel-based RNAi-producing systems.

描述（由申请人提供）：各种药物在血脑屏障（BBB）中的渗透性显着取决于位于脑毛细血管内皮细胞（BCEC）膜上的特定外排转运蛋白的表达和功能活性。 转运蛋白的选择性短暂下调将导致对CNS中脑肿瘤或病毒感染的更有效且毒性更少的治疗药物的应用。 以前，反义抑制剂已被证明可以暂时阻止主要的多药抗性剂，膜P-糖蛋白的合成，并促进抗性细胞表型的逆转。 最近已经发现了更有效的RNA干扰机制，用于关闭各种基因的表达。 短发夹RNA（siRNA）可以使用非病毒基因治疗方法通过质粒DNA前体引入靶细胞。 但是，将pDNA的靶向递送到BBB的细胞中需要一个良好的全身载体和与BCEC结合的选择性载体。 因此，通过脑特异性寄居肽（BSHP）修饰的聚合物交联的纳米凝胶颗粒已被选择通过用shrna对质粒DNA抑制特异性膜蛋白，药物污染物的特异性质子蛋白，在BBB中的特异性膜蛋白，在BBB中抑制质膜蛋白，以抑制特定的膜蛋白，在BBB中抑制BBB。 已从噬菌体显示库中的大量肽中选择了要连接到纳米凝胶表面和目标递送到BBB表面的特定BSHP。 纳米凝胶在许多细胞系中是无毒的，并且是转染剂的高效，显然是具有巨大潜力进行全身给药的载体之一。 矢量化的RNA干扰系统（RIP系统）可用于生物工程BBB渗透性，用于治疗剂，其大脑可及性受到特定药物外排转运蛋白的阻碍。 该提案的核心假设是，使用靶向的RNAI-生成系统通过全身转染BBB中选定的药物外反转运蛋白，可以在CNS相关疾病的化学疗法期间显着增强药物向大脑的药物转运。 我们的特定目的1是开发靶向BCEC的纳米凝胶载体，以便将质粒DNA全身递送到BBB。 具体目标2是增强BCEC靶向纳米凝胶载体在体外和体内的转染功效。 具体目的3是抑制体内BBB中选定的药物外运输蛋白，并暂时增加药物转运到大脑。 在此目标中，将在动物模型中评估几种具有核苷类模拟药物的大脑转运，因为基于纳米凝胶的RNAi-oproducing Systems在BBB中对药物外排转运蛋白的短暂下调。