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）， 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HIV-1 gp120和Tat等神经毒性病毒蛋白在大脑中的复制。抗逆转录病毒药物在穿过血脑屏障方面的效率各不相同，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。为了克服这一问题，我们建议采用模块化的、基于适配体的方法来提供旨在中和HIV-1和抑制隐藏在血脑屏障（BBB）后面的神经毒性病毒蛋白的治疗方法。我们将开发下一代递送剂，可以转糖血脑屏障，特异性靶向HIV，从而在中枢神经系统中获得更高的有效药物浓度，从而能够对抗和中和HIV-1及其有毒副产物。有两个具体目标。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CNS中的HIV库，该适体可以穿过由人脑微血管内皮细胞（HBMECs）组成的体外模型血脑屏障。利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，通过血脑屏障递送治疗性抗体、适体或其他分子，并尝试中和重组gp120或Tat蛋白介导的神经元杀伤或预防巨噬细胞的hiv感染。在Aim 2中，使用一种新颖的基于细胞的SELEX方法（通过指数扩增的配体选择性进化）和基于流动的动态体外血脑屏障（DIV-BBB; Flowcel Inc.），该方法在体内准确地再现了血脑屏障，我们寻求识别能够特异性穿过血脑屏障的RNA适体，并随后将其用于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.