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.

！1！ 摘要 该提案是对PA-19-237：病毒学(包括艾滋病毒)和免疫调节中的新RNA的回应： 基础科学和治疗发现。 HIV-1(HIV)感染CD4阳性细胞，导致获得性免疫缺陷综合征(AIDS)。多种形式的 细胞死亡(细胞凋亡、坏死、坏死性下垂、自噬、下垂、有丝分裂灾难等) 表明参与了病毒引起的直接感染细胞和旁观者细胞的丢失。当当前 现在，抗逆转录病毒疗法(ART)可以防止CD4的下降，并使大多数患者的CD4细胞数量恢复到接近正常的水平 在一些病例中，一个尚未解决的主要问题是为什么一些细胞在艾滋病毒感染后存活下来，而不是死亡，持续潜伏 普罗鲁斯。对艾滋病毒诱导细胞死亡机制的基本了解可以提供手段 杀死那些构成可激活病毒储存库的细胞，这些病毒决定了终身抗逆转录病毒。这样的细胞有 到目前为止，通过实验性的“电击并杀死”治愈策略避免了死亡。我们最近发现了一本小说 一种细胞死亡的形式，是到目前为止所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聚体种子毒性(6聚体种子毒素)是一种RNA干扰 基于(RNAi)的机制，通过针对反向互补种子匹配的有毒种子杀死细胞 在3‘非编码区中有大量对所有细胞的生存至关重要的基因。我们最近的数据表明 初级组织受到高度表达的miRNAs的保护，不受6聚体种子毒素的影响，这些miRNAs不携带有毒的 种子和阻止潜在有毒小RNA对RNA诱导的沉默复合体(RISC)的访问 介导RNAi。我们的新的初步数据表明，艾滋病毒感染会杀死缺乏这些保护的细胞 MiRNAs的效率要高得多，但它杀死无法形成功能RISC的细胞的效率较低 RNAi。这些数据表明，HIV诱导的细胞死亡涉及RNAi机制。第一个假设是 这一建议是，艾滋病毒通过启动6聚体种子毒素机制来杀死受感染的细胞，要么通过触发 产生细胞内源性有毒sRNA，或通过产生病毒编码的有毒sRNA进入RISC。 第二种假设是，HIV以一种导致miRNA下调的方式对感染细胞施加压力。 生物生成酶Disher降低保护性miRNAs的成熟度，使受感染的细胞对有毒的siRNAs敏感 它们是独立于迪特尔生成的。这些假设将在两个目标下进行研究：具体目标1： 确定HIV-1是否通过产生有毒病毒或在感染细胞中触发6聚体种子毒素 细胞sRNA。具体目标2：确定保护性miRNAs在两个方向上的调制是否呈现 HIV-1感染细胞对6聚体种子毒素有不同程度的敏感性。我们的工作将确定艾滋病毒是否会导致 通过6聚体种子毒性感染细胞。它可能通过以下方式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聚体种子毒素的诱导敏感，并减少手部神经元死亡。