Targeting CNS HIV reservoirs across the blood-brain barrier

描述(由申请人提供)：尽管进行了高度有效的抗逆转录病毒疗法(HAART)，但艾滋病毒相关神经疾病(HAND)的流行率仍在上升。HAND的发病机制主要是由于HIV-1gp120和Tat等神经毒性病毒蛋白在大脑中复制造成的。抗逆转录病毒药物在跨越血脑屏障方面具有不同的效率，因此无法消除中枢神经系统中的艾滋病毒-1或减少神经毒性病毒蛋白。为了克服这个问题，我们建议使用一种模块化的、基于适体的方法来提供治疗药物，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。我们将开发下一代递送剂，这些递送剂可以跨细胞转运BBB，专门针对艾滋病毒，从而在中枢神经系统实现更高的有效药物浓度，从而能够对抗和中和艾滋病毒-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能够穿过由人脑微血管内皮细胞(HBMECs)组成的体外模型BBB的抗转铁蛋白受体(HTfR)适配子来靶向TE CNS中的HIV储存库。利用这些适配子和我们的体外血脑屏障模型，我们将开发一种策略，在血脑屏障中传递治疗性抗体、适配子或其他分子，并试图中和重组gp120或Tat蛋白介导的神经元杀伤或防止巨噬细胞感染艾滋病毒。在目标2中，使用一种新的基于细胞的SELEX方法(通过指数扩增选择性进化配体)和使用基于流动的体外动态血脑屏障(DIV-BBB；Flowcel Inc.)为了在体内准确地复制BBB，我们试图识别能够特异性地穿过BBB的RNA适配子，并随后将这些适体调整为跨BBB的药物输送。

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.