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.

项目概要很明显，cART 治疗的 HIV 感染者中潜伏感染的 CD4+ T 细胞是主要的储存库本研究提案特别旨在解决题为“沉默”的 FOA。 HIV-1 Proviruses”，并由 FOA 提出以下具体目标：“利用 PIWI RNA 与其他小非编码 RNA 沉默途径（siRNA、shRNA、 miRNA），piRNA途径具有通过表观遗传永久沉默基因的理论优势除了 mRNA 降解活性之外，修饰（H3K9 甲基化），后者是主要模式其他小RNA的沉默，从而确保子代细胞中的基因沉默。表明 PIWI 蛋白在 CD4+ T 细胞和 T 细胞系中表达，并且似乎受到然而，PIWI-piRNA 通路是否可以被诱导发挥作用尚不清楚。我们发现 piRNA-PIWI 途径在体细胞中具有与在生殖细胞中相似的功能。在初级 CD4+ T 细胞中发挥作用，并且可以通过操纵来沉默 HIV。该项目的成功是由基础小非编码 RNA 和 piRNA 生物学领域的专家团队组成的 HIV 专家，在各自的领域都有长期的记录，我们将在提案的 R66 部分中测试证明。一个概念是，可以在 ACH-2 细胞中诱导针对 HIV 基因的 piRNA 途径，因此，当它们被细胞因子激活后，它们就停止产生 HIV 病毒体。如果成功，则病毒的 R33 部分。该提案将进一步描述 ACH-2 piRNA 的特征，这些 ACH-2 piRNA 具有针对广泛的 HIV 毒株的活性，包括取自 cART 治疗个体的离体原代细胞内的交叉进化枝作为临床前的一部分。目前的提案将探讨开发针对 CD4 受体的嵌合适体-piRNA 策略。将进一步揭示 piRNA 通路在人类细胞中的作用，以及它们是否可以被用作 HIV 沉默策略作为 HIV 治愈疗法的一部分。