Cell-derived extracellular vesicle mediated epigenetic silencing of HIV in the brain

项目摘要 细胞外囊泡介导的脑内HIV的表观遗传沉默 人类免疫缺陷病毒1型(HIV)是一种慢病毒，可导致持续性病毒感染和 导致免疫调节细胞死亡。通过免疫系统清除艾滋病毒感染是 低效，将前病毒整合到宿主细胞的基因组中提供了一种长期 持续性和潜伏期，需要终生抗逆转录病毒治疗。此外，这一点正变得越来越明显 HIV感染的单核/巨噬细胞是HIV-1在中枢神经系统的避难所 (中枢神经系统)，在那里它们似乎有助于艾滋病毒相关的神经疾病(手)。一个 可特异性靶向并在表观上沉默感染病毒内的艾滋病毒前病毒的方法学 大脑中的小胶质细胞可能是开发功能性治愈方法的一种手段，并可能是一种 手部的治疗。我们最近开发了一种锌指表观遗传抑制因子，它可以在表观遗传上 通过静脉注射细胞外小泡(EVS)使HIV在大脑中静默。我们在这里求婚 为了对比这种重组锌指方法EV方法和小发夹状RNA(ShRNA)EV方法 方法，这也是针对LTR，以表观遗传沉默艾滋病毒转录。的前提是 这项提议是，细胞衍生的电动汽车可以用来传递新型的抗艾滋病毒锌指或LTR 靶向转录调控shRNA到大脑，并在表观遗传上沉默艾滋病毒。我们建议3个 目的是验证细胞衍生受体靶向含有抗HIV锌的EV的假设 Finger和LTR指导的shRNA，两者都是利用内源性细胞的HIV转录调节因子 表观遗传沉默机制(3-5，14)，可在体内系统传播并稳定沉默HIV 抄写。我们将在体内使用模块化的胞外囊泡(EV)传递来验证这一假设 方法，通过神经干细胞(NSC)将被工程，使其结构性地生成 抗HIV EVS能够细胞导向稳定的表观遗传沉默HIV。如果成功，则该方法 这里概述的可能不仅导致HIV在大脑中的表观遗传沉默，而且还有助于迎来 可以与内源性细胞无缝操作的新一代EV-RNA疗法 针对基因转录的表观遗传调控的机制。

Project Summary Extracellular vesicle mediated epigenetic silencing of HIV in the brain Human Immunodeficiency Virus type 1 (HIV) is a lentivirus that causes a persistent viral infection and results in the demise of immune regulatory cells. Clearance of HIV infection by the immune system is inefficient, and integration of provirus into the genome of host cells provides a means for long-term persistence and latency which require lifelong anti-retroviral therapy. Moreover, it is becoming apparent that HIV-infected monocyte/macrophages represent a sanctuary for HIV-1 in central nervous system (CNS), where they appear to contribute to HIV-associated neurological disorders (HAND). A methodology that can specifically target and epigenetically silence HIV provirus within virus infected microglial cells in the brain could be one means by which to develop a functional cure and possibly a treatment for HAND. We recently developed a zinc finger epigenetic repressor that can epigenetically silence HIV in the brain when delivered by extracellular vesicles (EVs) intravenously. We propose here to contrast this recombinant zinc finger approach EV approach with a small hairpin RNA (shRNA) EV approach, which is also targeted to the LTR to epigenetically silence HIV transcription. The premise of this proposal is that cellular-derived EVs can be used to deliver novel anti-HIV zinc finger or LTR targeted transcriptional modulating shRNAs to the brain and epigenetically silence HIV. We propose 3 aims here to test the hypothesis that cellular derived receptor targeted EVs containing anti-HIV zinc finger and the LTR directed shRNA, both regulators of HIV transcription that utilize endogenous cellular epigenetic silencing mechanisms (3-5, 14), can spread systemically in vivo and stably silence HIV transcription. We will test this hypothesis here in vivo using a modular extracellular vesicle (EV) delivery approach, whereby by neural stem cells (NSC) will be engineered such that they constitutively generate anti-HIV EVs capable of cell directed stable epigenetic silencing of HIV. If successful the approach outlined here may not only result in the epigenetic silencing of HIV in the brain but also help usher in a new generation of EV-RNA therapies that can operate seamlessly with endogenous cellular mechanisms to target epigenetic regulation of gene transcription.