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.

项目摘要 细胞外囊泡介导的HIV脑内表观遗传沉默 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， 导致免疫调节细胞死亡。免疫系统清除HIV感染是 原病毒整合到宿主细胞的基因组中提供了一种长期的手段， 持久性和潜伏期，需要终身抗逆转录病毒治疗。此外，很明显， HIV感染单核/巨噬细胞是HIV-1在中枢神经系统中的避难所 (CNS)，在那里他们似乎有助于艾滋病毒相关的神经系统疾病（手）。一 可以特异性靶向和表观遗传学沉默病毒感染者体内的HIV前病毒的方法 大脑中的小胶质细胞可能是开发功能性治疗的一种手段， 手的治疗。我们最近开发了一种锌指表观遗传阻遏物， 当通过细胞外囊泡（EV）静脉内递送时，沉默脑中的HIV。我们在此提议 为了将这种重组锌指方法EV方法与小发夹RNA（shRNA）EV方法进行对比， 方法，其也靶向LTR以表观遗传学沉默HIV转录。的前提 这一提议是，细胞衍生的EV可用于递送新型抗HIV锌指或LTR 靶向转录调节shRNA到大脑和表观遗传沉默HIV。我们建议3 目的是检验细胞衍生受体靶向EV含有抗HIV锌的假设 finger和LTR指导的shRNA，两者都是HIV转录的调节因子， 表观遗传沉默机制（3-5，14），可在体内全身传播并稳定沉默HIV 转录。我们将在体内使用模块化细胞外囊泡（EV）递送来测试这一假设。 方法，其中神经干细胞（NSC）将被工程化，使得它们组成性地产生 抗HIV EV能够细胞定向的HIV的稳定表观遗传沉默。如果成功， 这里概述的可能不仅会导致大脑中HIV的表观遗传沉默，而且还有助于迎来一个新的艾滋病病毒感染者。 新一代EV-RNA疗法可以与内源性细胞 靶向基因转录的表观遗传调控机制。