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

细胞源性细胞外囊泡介导大脑中HIV的表观遗传沉默

• 批准号: 10748545
• 负责人: Fatah Kashanchi
• 金额: $ 63.14万
• 依托单位: GEORGE MASON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-02 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10748545
• 关键词: Acquired Immunodeficiency Syndrome; Adherence; Anti-Retroviral Agents; Astrocytes; Blood Vessels; Brain; CCR5 gene; CD34 gene; CXCR4 Receptors; Cell Line; Cell model; Cells; Central Nervous System; DNA Methylation; Death Rate; Disease; Drug Targeting; Drug resistance; Engineering; Enzymes; Epigenetic Process; Generations; Genes; Genetic Transcription; Genome; HIV; HIV Envelope Protein gp120; HIV Infections; HIV-1; Highly Active Antiretroviral Therapy; Human; Immune; Immune system; In Vitro; Individual; Infection; Intravenous; Lentivirus; Life Expectancy; Macrophage; Mediating; Meningeal; Methodology; Methods; Microglia; Modality; Morbidity - disease rate; Mus; Neurosphere; Nuclear Pore Complex; Opportunistic Infections; Pathway interactions; Patients; Persons; Pharmacotherapy; Provirus Integration; Proviruses; RNA; Recombinants; Repression; Shock; Site; Techniques; Testing; Therapeutic; Viral; Virus; Virus Diseases; Zinc Fingers; antiretroviral therapy; cost; epigenetic regulation; epigenetic silencing; extracellular vesicles; human disease; improved; in vivo; induced pluripotent stem cell; innovation; monocyte; mouse model; nerve stem cell; neuroAIDS; novel; particle; promoter; receptor; side effect; small hairpin RNA; therapeutic RNA

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.

项目概要 细胞外囊泡介导大脑中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，可引起持续性病毒感染并 导致免疫调节细胞死亡。免疫系统清除艾滋病毒感染的作用是 效率低下，将原病毒整合到宿主细胞基因组中提供了一种长期的手段 持续性和潜伏期需要终生抗逆转录病毒治疗。而且，越来越明显 感染 HIV 的单核细胞/巨噬细胞是中枢神经系统中 HIV-1 的避难所 (CNS)，它们似乎会导致 HIV 相关的神经系统疾病 (HAND)。一个 可以特异性靶向并表观遗传地沉默感染病毒内的 HIV 原病毒的方法 大脑中的小胶质细胞可能是开发功能性治疗方法的一种手段，也可能是一种 手部治疗。我们最近开发了一种锌指表观遗传阻遏物，可以表观遗传 通过细胞外囊泡 (EV) 静脉注射时，可抑制大脑中的 HIV。我们在这里提议 将这种重组锌指方法 EV 方法与小发夹 RNA (shRNA) EV 进行对比 该方法也针对 LTR，以表观遗传方式沉默 HIV 转录。前提是 该提议是细胞衍生的 EV 可用于传递新型抗 HIV 锌指或 LTR 将转录调节 shRNA 靶向大脑并通过表观遗传学沉默 HIV。我们建议3 这里的目的是检验细胞衍生受体靶向含有抗 HIV 锌的 EV 的假设 Finger 和 LTR 定向 shRNA，两者都是利用内源细胞的 HIV 转录调节因子 表观遗传沉默机制 (3-5, 14)，可以在体内系统性传播并稳定地沉默 HIV 转录。我们将使用模块化细胞外囊泡 (EV) 递送在体内测试这一假设 方法，通过神经干细胞（NSC）将被改造，使其组成型产生 抗HIV EVs能够对HIV进行细胞定向稳定表观遗传沉默。如果该方法成功 这里概述的可能不仅会导致大脑中艾滋病毒的表观遗传沉默，而且还有助于迎来 新一代 EV-RNA 疗法可与内源性细胞无缝协作 针对基因转录的表观遗传调控的机制。