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

基本信息

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
项目类别：
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

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

展开

项目摘要

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.

项目摘要细胞囊泡介导的大脑中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，会引起持续的病毒感染和导致免疫调节细胞的消亡。免疫系统清除HIV感染的是效率低下，并且将病毒病毒整合到宿主细胞的基因组中，为长期提供了一种手段持久性和潜伏期需要终身抗逆转录病毒疗法。而且，它变得显而易见感染HIV的单核细胞/巨噬细胞代表了中枢神经系统中HIV-1的避难所（CNS），它们似乎有助于与HIV相关的神经系统疾病（手）。一个可以在病毒感染中特异性靶向和表观遗传沉默的HIV病毒的方法论大脑中的小胶质细胞可能是开发功能治愈的一种手段，可能是手的治疗。我们最近开发了一个锌指表观遗传阻遏物，可以表观遗传通过静脉注射细胞外囊泡（EV）传递大脑中的艾滋病毒。我们在这里提出建议为了将这种重组锌指方法与小发夹RNA（shRNA）eV进行对比方法也针对LTR到表观遗传上的沉默HIV转录。前提该建议是，可以使用细胞来源的电动汽车来传递新型的抗HIV锌指或LTR 针对大脑的靶向转录调节shRNA和表观遗传沉默的HIV。我们提出3 目的是检验以下假设：含有抗HIV锌的细胞衍生受体靶向EV 手指和LTR的shRNA，这是使用内源细胞的HIV转录的调节剂表观遗传沉默机制（3-5，14）可以在体内全身传播，稳定的HIV稳定转录。我们将在这里使用模块化细胞外囊泡（EV）递送在体内检验该假设方法，通过神经干细胞（NSC）进行设计，以使它们构成生成抗HIV电动汽车，能够定向稳定的艾滋病毒表观遗传沉默。如果成功的方法这里概述的不仅可能导致大脑中艾滋病毒的表观遗传沉默，还可以帮助迎接新一代的EV-RNA疗法，可以用内源性细胞无缝运行靶向基因转录表观遗传调节的机制。