Innate Sensing of Cell-Free DNA and the Interferon-Mediated Control of HIV In Vivo

项目摘要 抗逆转录病毒治疗（ART）的出现大大降低了发病率和死亡率， 与HIV感染相关，由于潜伏感染细胞在治疗期间的持续存在， 治疗越来越多的数据表明，艾滋病毒感染者经常经历持续的免疫反应， 即使在ART介导的病毒感染的情况下， 镇压这些现实使人们对制定治疗艾滋病毒感染的战略产生了明显的兴趣。 确定艾滋病毒持续存在的宿主分子决定因素和停药后的反弹动力学 抗逆转录病毒疗法（ART）将是制定有效的策略，治愈艾滋病毒感染的关键。我们最近 对来自HIV感染者的血浆样本应用了一种全面的系统分析方法， 接受分析治疗中断（ATI），以确定能够预测 HIV在ART中断后的反弹动力学，并作为潜在的药理学靶点，以促进 在没有抗逆转录病毒治疗的情况下，持久的病毒学缓解。 与ART停止后HIV反弹时间相关的是循环游离DNA（cfDNA）， 等离子体具体地，血浆中cfDNA丰度增加与延迟的反弹时间相关。 在程序性细胞死亡期间释放到循环中的cfDNA已经在生物医学领域中被广泛利用。 临床肿瘤学（通常称为“液体活检”）。然而，cfDNA仍然在很大程度上未被探索的背景下， 艾滋病毒感染。在R 01提案中，我们严格探索cfDNA作为HIV疾病状态的生物标志物， 研究cfDNA与病毒表达和反弹的分子和免疫机制。 我们的中心假设是：1）cfDNA反映了体内HIV感染细胞的死亡率;以及2） cfDNA促进宿主中的抗病毒状态（例如诱导I型干扰素应答）， 防止病毒反弹我们的项目具有基础，翻译和临床研究团队 它利用了来自特征明确的艾滋病毒感染者的大量临床样本。 在目标1中，我们将应用下一代测序（NGS）方法对来自HIV感染者的血浆样本进行测序。 验证和增强cfDNA作为自然HIV控制生物标志物的预后意义 in vivo.在目标2中，我们将评估HIV靶细胞对cfDNA的摄取和感知如何影响细胞命运。 以及艾滋病毒的重新激活和复制。在目标3中，我们将确定HIV相关的线粒体 功能障碍导致cfDNA挤出并最终诱导I型干扰素应答， 反弹我们的项目将推动艾滋病毒治疗策略的开发和评估。

PROJECT SUMMARY While the advent of antiretroviral therapy (ART) has dramatically reduced the morbidity and mortality associated with HIV infection, a cure is not achieved due to the persistence of latently-infected cells during treatment. Accumulating data suggest that HIV-infected individuals often experience persistent immune dysregulation, chronic inflammation, and accelerated aging even in the setting of ART-mediated viral suppression. These realities have created a pronounced interest in developing strategies to cure HIV infection. Identifying the host molecular determinants of HIV persistence and rebound kinetics following cessation of antiretroviral therapy (ART) will be critical in developing effective strategies to cure HIV infection. We recently applied a comprehensive systems profiling approach to plasma samples from HIV-infected individuals who underwent analytical treatment interruption (ATI), to identify circulating host factors that enable prediction of HIV rebound kinetics following ART interruption, and serve as potential pharmacological targets to promote durable virologic remission in the absence of ART. The host factor exhibiting the strongest statistical association with HIV rebound timing following ART cessation was circulating cell-free DNA (cfDNA) in plasma. Specifically, increased cfDNA abundance in plasma was associated with delayed time-to-rebound. cfDNA, released into circulation during programmed cell death, has been exploited extensively in the realm of clinical oncology (often termed “liquid biopsy”). However, cfDNA remains largely unexplored in the setting of HIV infection. In this R01 proposal, we rigorously explore cfDNA as a biomarker of HIV disease states, and we investigate the molecular and immunologic mechanisms linking cfDNA to viral expression and rebound. Our central hypotheses are that 1) cfDNA reflects the death rate of HIV-infected cells in vivo; and 2) cfDNA promotes an antiviral state in the host (e.g. induction of type I interferon response) which prevents viral rebound. Our project features an investigative team with basic, translational, and clinical expertise, and leverages large collections of clinical samples from well-characterized HIV-infected individuals. In Aim 1, we will apply next-generation sequencing (NGS) approaches to plasma samples from HIV-infected individuals to validate and enhance the prognostic significance of cfDNA as a biomarker of natural HIV control in vivo. In Aim 2, we will evaluate how the uptake and sensing of cfDNA by HIV target cells impacts cell fate and the reactivation and replication of HIV. In Aim 3, we will determine how HIV-associated mitochondrial dysfunction leads to cfDNA extrusion and ultimately induces type I interferon responses that prevent HIV rebound. Our project will advance the development and evaluation of HIV cure strategies.