Targeting CNS HIV reservoirs across the blood-brain barrier

跨越血脑屏障靶向中枢神经系统艾滋病毒储存库

• 批准号: 8544744
• 负责人: MATTHEW LEVY
• 金额: $ 25.42万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-25 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8544744
• 关键词: Acids; Animal Model; Anti-HIV Agents; Antibodies; Biological Assay; Blood - brain barrier anatomy; Blood Circulation; Blood capillaries; Brain; Brain Diseases; Cells; Chemistry; Clinic; Collaborations; Drug Combinations; Drug Delivery Systems; Endothelial Cells; Evolution; Exhibits; Future; HIV; HIV Antibodies; HIV Envelope Protein gp120; HIV Infections; HIV therapy; HIV-1; Health; Highly Active Antiretroviral Therapy; Human; In Vitro; Infection; Inflammatory; Laboratories; Length; Ligands; Mediating; Modality; Modeling; Morbidity - disease rate; Neurons; Neurotoxins; Pathogenesis; Patients; Penetration; Pharmaceutical Preparations; Prevalence; Property; Proteins; RNA; Recombinants; Research; Resistance; Role; Route; System; Testing; Therapeutic; Therapeutic antibodies; Transferrin; Transferrin Receptor; Variant; Viral Proteins; Virion; aptamer; base; capillary; chemokine; combat; cytokine; design; improved; in vitro Model; in vivo; killings; macrophage; monolayer; nervous system disorder; neurotoxic; neurotoxicity; neutralizing antibody; next generation; novel; nuclease; prevent; public health relevance; research study; tat Protein; therapeutic target

DESCRIPTION (provided by applicant): The prevalence of HIV-associated neurological disorders (HAND) is on the rise despite Highly Active Anti-Retroviral Therapy (HAART). The pathogenesis of HAND is primarily due to neurotoxic viral proteins such as HIV-1 gp120 and Tat due to the replication of HIV in the brain. Antiretrovirals have variable efficiencies in crossing he blood brain barrier and thus being unable to eliminate HIV-1 in the CNS or in reducing the neurotoxic viral proteins. To overcome this problem, we propose to employ a modular, aptamer-based approach to deliver therapeutics designed to neutralize HIV-1 and inhibit neurotoxic viral proteins hiding behind the blood brain barrier (BBB). We will develop the next generation of delivery agents that can transcytose the BBB, specifically target HIV and consequently achieve higher effective drug concentrations in the CNS to be able to combat and neutralize HIV-1 and its toxic byproducts. There will be two specific aims. In Aim 1, we will target HIV reservoirs in te CNS using previously selected anti-transferrin receptor (hTfR) aptamers that can cross an in vitro model BBB composed of Human Brain Microvascular Endothelial Cells (HBMECs). Using these aptamers and our in vitro BBB model, we will develop a strategy to deliver therapeutic antibodies, aptamers or other molecules across the BBB and attempt to neutralize recombinant gp120 or Tat protein-mediated neuronal killing or to prevent HIV-infection of macrophages. In Aim 2, using a novel cell-based SELEX approach (selective evolution of ligands by exponential amplification) and using a flow-based dynamic in vitro blood brain barrier (DIV-BBB; Flowcel Inc.) which accurately reproduces the BBB in vivo, we seek to identify RNA aptamers that can specifically cross the BBB and will subsequently adapt these for drug delivery across the BBB.

描述（由申请人提供）：尽管具有高度活跃的抗逆转录病毒疗法（HAART），与HIV相关的神经系统疾病（HARD）的患病率正在上升。手的发病机理主要是由于神经毒性病毒蛋白（例如HIV-1 GP120）和TAT引起的，这是由于大脑中HIV的复制而引起的。抗逆转录病毒在跨越血脑屏障的效率上有可变的效率，因此无法消除中枢神经系统中的HIV-1或减少神经毒性病毒蛋白。为了克服这个问题，我们建议采用一种基于适体的模块化方法来提供旨在中和HIV-1并抑制隐藏在血脑屏障（BBB）后面的神经毒性病毒蛋白。我们将开发下一代可以转化BBB的输送剂，特别是针对HIV，因此在中枢神经系统中获得了更高的有效药物浓度，以便能够对抗和中和HIV-1及其有毒的副产品。将有两个具体的目标。在AIM 1中，我们将使用先前选择的抗转铁蛋白受体（HTFR）适体靶向TE CNS中的HIV储存剂，该适体可以穿越由人脑微血管内皮细胞（HBMEC）组成的体外模型BBB。使用这些适体和我们的体外BBB模型，我们将制定一种策略，以跨BBB提供治疗性抗体，适体或其他分子，并试图中和重组GP120或TAT蛋白质介导的神经元杀伤或防止巨噬细胞HIV的感染。在AIM 2中，使用一种新型的基于细胞的SELEX方法（通过指数扩增对配体进行选择性演变），并使用基于流动的动态体外血脑屏障（Div-BBB； Flowcel Inc.），我们可以准确地重现BBB INVIVO，我们试图确定可以特异性地跨越BBB的RNA Aptamer，并将这些aptams识别为bbb和随后的药物进行适应bbb。