6mer seed toxicity and AIDS

6mer 种子毒性和艾滋病

• 批准号: 10132980
• 负责人: Marcus E. Peter
• 金额: $ 23.85万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-25 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10132980
• 关键词: 3' Untranslated Regions; Acquired Immunodeficiency Syndrome; Antioxidants; Apoptosis; Automobile Driving; Autophagocytosis; Basic Science; Biogenesis; CD4 Positive T Lymphocytes; Cell Death; Cell Death Induction; Cell Line; Cells; Cessation of life; Code; Complex; Data; Data Set; Dicer Enzyme; Down-Regulation; Generations; Genes; Goals; HIV; HIV Infections; HIV-1; Hybrids; Infection; Knock-out; Ligation; Mediating; Messenger RNA; MicroRNAs; Mitotic; Molecular; Necrosis; Pathway interactions; Pharmacotherapy; Production; Proteins; Proviruses; RNA; RNA Interference; RNA-Induced Silencing Complex; Reactive Oxygen Species; Route; Seeds; Shock; Site; Small Interfering RNA; Small RNA; Stress; T-Lymphocyte; Testing; Therapeutic; Tissues; Toxic effect; Untranslated RNA; Viral; Virus; Virus Replication; Work; antiretroviral therapy; base; cell killing; crosslink; effective therapy; immunoregulation; in vivo; knock-down; mutant; neuron loss; novel; overexpression; prevent; response; transcriptome sequencing; virology

! 1! Summary The proposal is in response to PA-19-237: Novel RNAs in Virology (including HIV) and Immune Regulation: Basic Science and Therapeutic Discovery. HIV-1 (HIV) infects CD4 positive cells causing acquired immunodeficiency syndrome (AIDS). Many forms of cell death (apoptosis, necrosis, necroptosis, autophagy, pyroptosis, mitotic catastrophe, and others) have been shown to be involved in virus induced cell loss of directly infected and bystander cells. While current antiretroviral therapy (ART) now prevents CD4 decline and restores their numbers to nearly normal in most cases, a major unsolved problem is why some cells survive HIV infection rather than dying, persist with latent provirus. A fundamental understanding of mechanisms of cell death induction by HIV could provide the means to kill those cells that constitute a reservoir of reactivatable virus that mandates lifelong ART. Such cells have so far evaded death from experimental “shock and kill” cure strategies. We have recently discovered a novel form of cell death that is a combination of almost all of the above-mentioned mechanisms implicated to date in cell death associated with untreated HIV infection. 6mer Seed Toxicity (6mer Seed Tox) is an RNA interference (RNAi) based mechanism that kills cells through toxic seeds that target reverse complementary seed matches in the 3'UTR of a large number of genes that are critical for the survival of all cells. Our recent data suggest that primary tissues are protected from 6mer Seed Tox by highly expressed miRNAs that do not carry a toxic seed and block access of the potentially toxic small RNAs to the RNA induced silencing complex (RISC) that mediates RNAi. Our new preliminary data demonstrate that HIV infection kills cells that lack these protective miRNAs much more efficiently and it kills cells less efficiently that cannot form a functional RISC to mediate RNAi. These data suggest that cell death induced by HIV involves the RNAi machinery. The first hypothesis of this proposal is that HIV kills infected cells by engaging the 6mer Seed Tox mechanism either by triggering the generation of cell-endogenous toxic sRNAs or by producing virus-encoded toxic sRNAs that enter the RISC. The second hypothesis is that HIV stresses infected cells in a way that causes downregulation of the miRNA biogenesis enzyme Dicer decreases maturation of protective miRNAs, sensitizing infected cells to toxic siRNAs that are generated independently of Dicer. These hypotheses will be studied in two aims: Specific Aim 1: Determine whether HIV-1 triggers 6mer Seed Tox in infected cells through the production of toxic viral or cellular sRNAs. Specific Aim 2: Determine whether modulation of protective miRNAs in either direction renders HIV-1 infected cells more or less susceptible to 6mer Seed Tox. Our work will establish whether HIV kills infected cells through 6mer seed toxicity. It may pave the way to advance current HIV eradication strategies by sensitizing cells to induction of 6mer Seed Tox after latency reversal and decreasing neuronal death in HAND.

!一个！ 总结 该提案是对PA-19-237：病毒学（包括HIV）和免疫调节中的新型RNA的回应： 基础科学和治疗发现。 HIV-1（HIV）感染CD 4阳性细胞，导致获得性免疫缺陷综合征（AIDS）。许多形式的 细胞死亡（凋亡、坏死、坏死性凋亡、自噬、焦亡、有丝分裂灾难等）已被 显示参与病毒诱导的直接感染细胞和旁观者细胞的细胞损失。虽然目前的 抗逆转录病毒疗法（ART）现在可以防止CD 4下降，并使大多数人的CD 4数量恢复到接近正常水平。 在许多病例中，一个尚未解决的主要问题是为什么有些细胞在艾滋病毒感染后存活而不是死亡， 前病毒对HIV诱导细胞死亡机制的基本理解可以提供一种方法， 杀死那些构成可再活化病毒库的细胞，这些病毒库要求终身进行ART。 迄今为止，从实验性的“休克和杀死”治疗策略中逃脱了死亡。我们最近发现了一本小说 细胞死亡的一种形式，它是迄今为止涉及的几乎所有上述机制的组合， 与未经治疗的艾滋病毒感染有关的细胞死亡。6 mer种子毒性（6 mer Seed Tox）是一种RNA干扰 基于RNAi的机制，通过靶向反向互补种子匹配的毒性种子杀死细胞 在对所有细胞的存活至关重要的大量基因的3 'UTR中。我们最近的数据显示 原代组织通过高表达的miRNAs保护不受6聚体种子毒素的影响， 接种并阻断潜在毒性的小RNA进入RNA诱导沉默复合物（RISC）， 介导RNAi。我们新的初步数据表明，艾滋病毒感染杀死缺乏这些保护性的细胞， 它杀死不能形成功能性RISC来介导的细胞的效率较低 RNA干扰这些数据表明，HIV诱导的细胞死亡涉及RNAi机制。第一个假设 该提议是艾滋病毒通过启动6 mer种子毒性机制或通过触发 产生细胞内源性毒性sRNA或通过产生进入RISC的病毒编码的毒性sRNA。 第二个假设是HIV以一种方式胁迫感染细胞，导致miRNA下调 生物合成酶Dicer降低保护性miRNA的成熟，使感染细胞对毒性siRNA敏感 这些都是独立于切丁酶产生的。这些假设将在两个目标中进行研究：具体目标1： 确定HIV-1是否通过产生毒性病毒或毒素在感染细胞中触发6聚体种子毒性， 细胞sRNAs。具体目标2：确定在任一方向上调节保护性miRNAs是否会导致 HIV-1感染的细胞或多或少对6聚体种子毒性敏感。我们的工作将确定艾滋病毒是否会杀死 通过6 mer种子毒性感染细胞。它可能为推进当前的艾滋病毒根除战略铺平道路， 在潜伏期逆转后使细胞对6聚体种子Tox的诱导敏感，并减少HAND中的神经元死亡。