Harnessing the PIWI piRNA pathway to silence HIV

利用 PIWI piRNA 途径抑制 HIV

基本信息

批准号：
9762886
负责人：
Mario Ostrowski
金额：
$ 44.1万
依托单位：
UNIVERSITY OF TORONTO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-15 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9762886
关键词：
Address Antiviral Therapy Biology Blood CD4 Antigens CD4 Positive T Lymphocytes CRISPR/Cas technology Cell Line Cell model Cells Cytokine Activation Ensure Epigenetic Process Family Gene Silencing Gene Silencing Pathway Genes Germ Cells HIV HIV Infections HIV-1 Human Human immunodeficiency virus test Individual Knowledge Length Life Methylation MicroRNAs Modeling Modification Ovary Pathway interactions Persons Preparation Proteins Proviruses RNA Interference Records Research Proposals Retroelements Role Small Interfering RNA Small RNA Somatic Cell Stem cells T-Lymphocyte Testing Testis Tissues Transgenic Organisms Untranslated RNA Virion Virus Work aptamer cohort cytokine design embryo cell high risk immune activation in vivo insight mRNA Transcript Degradation member memory CD4 T lymphocyte novel novel therapeutic intervention piRNA pre-clinical preclinical development programs small hairpin RNA success

项目摘要

PROJECT SUMMARY It is clear that latently infected CD4+ T cells in cART treated HIV-infected individuals are the major reservoir and obstacle to eradication. This research proposal specifically intends to address the FOA entitled “Silencing of HIV-1 Proviruses” with the following specific objective proposed by the FOA: “ to harness the PIWI RNA pathway to silence HIV”. In contrast to other small non-coding RNA silencing pathways (siRNA, shRNA, miRNA), the piRNA pathway has the theoretical advantage of permanently silencing genes through epigenetic modifications (H3K9 methylation) in addition to mRNA degradation activity, the latter being the primary mode of silencing by other small RNAs, thus, ensuring gene silencing in progeny cells. Our preliminary findings indicate that PIWI proteins are expressed in CD4+ T cells and T cell lines, and appear to be regulated by immune activation. It is unknown, however whether the PIWI-piRNA pathway can be induced to perform similar functions in somatic cells as they do in germ cells. We hypothesize that the piRNA-PIWI pathway is functional in primary CD4+ T-cells and can be manipulated to silence HIV. Attempts will be made to ensure success of the project by teaming experts in fundamental small noncoding RNA and piRNA biology with an HIV expert, all having long track records in their fields. In the R66 portion of the proposal we will test the proof of concept that that one can induce a piRNA pathway against HIV genes in ACH-2 cells, so that, when they are activated by cytokine, they have ceased to produce HIV virions. If successful, the R33 portion of the proposal will further characterize ACH-2 piRNAs that have activity against a broad range of HIV strains, including cross Clades within ex vivo primary cells taken from cART treated individuals. As part of pre-clinical development a CD4 receptor targeting chimeric aptamer-piRNA strategy will be explored. The current proposal will reveal further insights on the role of the piRNA pathway in human cells, whether they can be exploited as an HIV silencing strategy as part of the armamentarium toward HIV cure therapy.

项目摘要很明显，在被接受的艾滋病毒感染的个体中，受到潜在感染的CD4+ T细胞是主要的研究者和根除的障碍。这项研究建议专门打算解决标题为“无声的FOA” FOA提出的以下特定目标：“利用Piwi RNA 与其他小型非编码RNA沉默途径相比（sirna，shrna， miRNA），piRNA途径具有通过表观遗传学永久沉默基因的理论优势除mRNA降解活性外，修饰（H3K9甲基化），后者是主要模式因此，由其他小的RNA沉默，以确保进度细胞中的基因沉默。我们的初步发现表明PIWI蛋白在CD4+ T细胞和T细胞系中表达，并且似乎受免疫激活。这是未知的，但是是否可以诱导Piwi-piRNA途径执行在体细胞中的功能类似，就像在生殖细胞中一样。我们假设Pirna-Piwi途径是在原代CD4+ T细胞中发挥作用，可以操纵以沉默HIV。将尝试确保通过将基本小型非编码RNA和PIRNA生物学的专家组成的专家与艾滋病毒专家，在自己的领域都有长期记录。在提案的R66部分中，我们将测试证明人们可以在ACH-2细胞中诱导针对HIV基因的PIRNA途径的概念，因此，当它们被细胞因子激活，它们已经停止产生HIV病毒。如果成功，则R33部分提案将进一步表征ACH-2 PIRNA具有针对广泛艾滋病毒菌株的活性，包括从接受过的手推车的个体中取出的离体主细胞内的跨进枝。作为临床前的一部分开发将探讨CD4受体靶向嵌合磷脂策略。当前的建议将揭示对PIRNA途径在人类细胞中的作用的进一步见解，是否可以探讨它们 HIV沉默策略是艾滋病毒治疗疗法的武器库的一部分。