Silencing of HIV-1 proviruses by epigenetically targeting transcriptional elongation

通过表观遗传靶向转录延伸来沉默 HIV-1 原病毒

• 批准号: 10205410
• 负责人: Qin Feng
• 金额: $ 38.71万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10205410
• 关键词: AIDS prevention; Alkaloids; Baculoviruses; Binding; Biological Assay; Cell model; Cells; Chemistry; Combined Modality Therapy; Complex; DNA; Disease; Drug Kinetics; Economics; Epigenetic Process; Gene Expression; Genes; Genetic Transcription; Goals; HIV; HIV Infections; HIV-1; Hepatitis C virus; Herpesviridae; Human; Human Herpesvirus 4; Immunotherapy; Infection; Interruption; Knowledge; Life; Names; Nature; Patients; Peptide Sequence Determination; Pharmaceutical Preparations; Phase; Positive Transcriptional Elongation Factor B; Production; Proteins; Proviruses; Reader; Recurrence; Rest; Shock; Steroids; Technology; Testing; Therapeutic; Tissues; Toxic effect; Transcription Elongation; Translating; Treatment Efficacy; Validation; Viral; Virus; Virus Latency; Virus Replication; analog; antiretroviral therapy; base; candidate identification; clinical application; cortistatin; epigenetic regulation; inhibitor/antagonist; latent HIV reservoir; memory CD4 T lymphocyte; novel; novel therapeutics; prevent; purge; reactivation from latency; screening; side effect; small molecule; small molecule inhibitor; tat Protein; therapeutic target

Project Summary/Abstract Combination Antiretroviral Therapy (cART) has been tremendously effective in HIV-infected patients by suppressing HIV replication and maintaining the virus at undetectable levels. However, although cART is life- prolonging, it never eradicates HIV infection or cures this disease. Interruption of treatment quickly results in virus replication and recurrence of the disease. As a result, HIV-infected patients have to receive lifelong cART treatment which presents economic and therapeutic burdens. To this end, a “shock and kill” strategy has been proposed to reactivate (“shock”) latent HIV in the presence of cART and immune-based therapy to purge (“kill”) these reservoirs. However several serious issues surrounding this strategy limit its clinical application. Therefore there is an urgent need to develop novel effective therapeutic strategy. Recently a “soothe and snooze” strategy has been proposed to permanently silence the latent provirus by stronger and more durable virus-suppressing agents. Didehydro-cortistatin A (dCA), a specific inhibitor for HIV-1 Tat, a key HIV viral trans-activating regulatory protein, has been shown to prevent HIV-1 reactivation from latency. In this project, we seek to develop novel and more effective drugs to silence the latent HIV provirus, based on our knowledge on epigenetic regulation of HIV gene transcription. It has been established that elongation is the rate limiting step of HIV gene transcription. We hypothesize that inhibition of elongation will block HIV provirus gene expression, resulting in deep viral suppression. The super elongation complex (SEC) is critical for HIV transcription elongation. In our preliminary study, we have shown that the core component of the SEC, the AFF proteins, contain novel epigenetic ‘reader and ‘writer’ (HAT) activities, making them ideal therapeutic targets. We will apply the cutting edge DNA-encoded chemistry technology (DEC-Tec) to perform large scale screening to identify small molecule inhibitors of AFF1 and AFF4. Such inhibitors would silence the HIV provirus, and combination of AFF inhibitors and Tat inhibitor dCA might further suppress HIV activation to achieve a functional cure for HIV-1 infection. Our proposal will be carried in two phases. In R61 phase of the first three years, the goal is to identify AFF1 and AFF4 inhibitors. Aim 1, to identify small-molecule compounds that bind to AFF1/4 by DNA-encoded chemistry technology. Aim 2, to test if these compounds inhibit HIV transcription and AFF function. In R33 phase of the last two years, the goal is to perform functional characterization of AFF1/4 inhibitors. Aim 3, validation of inhibitory activity of AFF1/4 inhibitors on HIV transcription in primary cell model and combination therapy with Tat inhibitor dCA. Aim 4, pharmacokinetics (PK) and toxicity studies of AFF1/4 inhibitors.

项目总结/摘要 联合抗逆转录病毒疗法（cART）在HIV感染患者中非常有效， 抑制HIV复制并将病毒维持在无法检测的水平。然而，尽管cART是生命- 长期以来，它从未根除艾滋病毒感染或治愈这种疾病。治疗中断很快会导致 病毒复制和疾病复发。因此，艾滋病毒感染者必须终身接受 cART治疗带来经济和治疗负担。为此，采取了“震慑”策略 已经提出在cART和基于免疫的治疗存在下重新激活（“休克”）潜伏的HIV， 清除（“杀死”）这些水库。然而，围绕该策略的几个严重问题限制了其临床应用。 应用程序.因此，迫切需要开发新的有效的治疗策略。 最近提出了一种“安抚和打盹”策略，通过以下方式永久沉默潜伏的前病毒： 更强和更持久的病毒抑制剂。二脱氢皮质抑素A（dCA），一种特异性抑制剂， HIV-1达特是HIV病毒反式激活的关键调节蛋白，已被证明可阻止HIV-1的再激活 从latency。在这个项目中，我们寻求开发新的和更有效的药物来沉默潜伏的艾滋病毒 前病毒，基于我们对HIV基因转录的表观遗传调控的知识。 已经确定延伸是HIV基因转录的限速步骤。我们假设 延长的抑制将阻断HIV前病毒基因表达，导致深度病毒抑制。超级 延伸复合物（SEC）对于HIV转录延伸至关重要。在我们的初步研究中， SEC的核心成分AFF蛋白含有新的表观遗传“阅读器和”写入器“（HAT）， 使其成为理想的治疗目标。我们将运用尖端的DNA编码化学 技术（DEC-Tec）进行大规模筛选，以鉴定AFF 1的小分子抑制剂， AFF4。这种抑制剂将使HIV前病毒沉默，AFF抑制剂和达特抑制剂的组合 dCA可能进一步抑制HIV活化，以实现HIV-1感染的功能性治愈。 我们的建议将分两个阶段进行。在前三年的R61阶段，目标是识别AFF 1 AFF 4抑制剂目的1，鉴定通过DNA编码的结合AFF 1/4的小分子化合物， 化学技术目的2：检测这些化合物是否抑制HIV转录和AFF功能。在R33中 在过去两年的第一阶段，目标是对AFF 1/4抑制剂进行功能表征。目标3， 在原代细胞模型和组合中验证AFF 1/4抑制剂对HIV转录的抑制活性 用达特抑制剂dCA治疗。目的4：AFF 1/4抑制剂的药代动力学（PK）和毒性研究。