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.

呢1！ 概括 该提案是对PA-19-237的回应：病毒学中的新RNA（包括艾滋病毒）和免疫调节： 基础科学和治疗发现。 HIV-1（HIV）感染CD4阳性细胞引起获得性免疫缺陷综合征（AIDS）。多种形式 细胞死亡（细胞凋亡，坏死，坏死，自噬，凋亡，有丝分裂灾难等） 证明参与病毒引起的直接感染和旁观者细胞的细胞丧失。当前 现在，抗逆转录病毒疗法（ART）可防止CD4下降，并将其数量恢复到大多数人的数量正常 案例，一个主要的未解决的问题是为什么某些细胞在HIV感染而不是死亡的情况下持续存在。 病毒。对艾滋病毒诱导细胞死亡机制的基本理解可以提供手段 杀死那些构成命令终身艺术的可重复病毒储层的细胞。这些细胞具有 到目前为止，从实验性的“冲击和杀死”治愈策略中逃避了死亡。我们最近发现了一本小说 细胞死亡的形式是迄今为止几乎所有上述机制的组合 与未处理的HIV感染相关的细胞死亡。 6mer种子毒性（6mer种子托克斯）是RNA干扰 基于（RNAi）通过有毒种子杀死细胞的基于（RNAi）的机制 在大量基因的3'UTR中，这对于所有细胞的存活至关重要。我们最近的数据暗示 原代组织受到高度表达的miRNA的保护，不携带有毒的miRNA 种子和阻止潜在有毒的小RNA进入RNA引起的沉默复合物（RISC） 介导RNAi。我们的新初步数据表明，艾滋病毒感染杀死缺乏这些受保护的细胞 miRNA更有效地杀死细胞的效率较低，无法形成功能性RISC以介导 RNAi。这些数据表明，HIV诱导的细胞死亡涉及RNAi机械。第一个假设 该建议是，艾滋病毒通过触发6mer种子托克斯机制来杀死受感染的细胞 生成细胞源性毒性SRNA或通过产生进入RISC的病毒编码的有毒sRNA。 第二个假设是，艾滋病毒以导致miRNA下调的方式将受感染的细胞压力 生物发生酶dicer降低了受保护的miRNA的成熟，使感染细胞对有毒siRNA敏感 独立于DICER产生的。这些假设将以两个目的进行研究：特定目标1： 确定HIV-1是否通过产生有毒病毒或 细胞SRNA。特定目标2：确定是否在任一方向上调节受保护的miRNA HIV-1感染的细胞或多或少容易受到6mer种子托克斯的影响。我们的工作将确定艾滋病毒是否杀害 通过6mer种子毒性感染细胞。它可能为通过 在潜伏期逆转后，对细胞诱导6mer种子毒物的敏感性减少了手头的神经元死亡。