Targeting CNS HIV reservoirs across the blood-brain barrier

描述（由申请人提供）：尽管进行了高效抗逆转录病毒治疗（HAART），但HIV相关神经系统疾病（HAND）的患病率仍在上升。HAND的发病机制主要是由于神经毒性病毒蛋白，如HIV-1 gp 120和达特，由于HIV在脑中的复制。抗逆转录病毒药物在穿过血脑屏障方面具有不同的效率，因此不能消除CNS中的HIV-1或减少神经毒性病毒蛋白。为了克服这个问题，我们建议采用一种模块化的，基于适体的方法来提供治疗，旨在中和HIV-1和抑制隐藏在血脑屏障（BBB）后面的神经毒性病毒蛋白。我们将开发下一代的传递剂，可以transcytose血脑屏障，专门针对艾滋病毒，从而实现更高的有效药物浓度在中枢神经系统，能够打击和中和艾滋病毒-1及其有毒副产物。将有两个具体目标。在目标1中，我们将使用先前选择的抗转铁蛋白受体（hTfR）适体靶向CNS中的HIV储库，所述适体可以穿过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血脑屏障模型，我们将开发一种策略，以提供治疗性抗体，适体或其他分子通过血脑屏障，并试图中和重组gp 120或达特蛋白介导的神经元杀伤或防止艾滋病毒感染的巨噬细胞。在目的2中，使用新的基于细胞的SELEX方法（通过指数扩增的配体的选择性进化）和使用基于流动的动态体外血脑屏障（DIV-BBB; Flowcel Inc.）其在体内准确地复制血脑屏障，我们试图鉴定可以特异性地穿过血脑屏障的RNA适体，并随后使这些适体适应于穿过血脑屏障的药物递送。

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.