Martin Delaney Collaboratory to Eradicate HIV-1 Infection

描述(由申请人提供)：尽管抗逆转录病毒疗法(ART)在临床上取得了成功，但每天感染人类免疫缺陷病毒(HIV)的人比开始抗逆转录病毒治疗的人更多。终身抗逆转录病毒治疗的困难--特别是在发展中国家--使根除艾滋病毒成为当务之急。但对于接受抗逆转录病毒治疗的患者来说，清除逆转录病毒感染是一项艰巨的任务。尽管人们对艾滋病的持续存在有很多了解，但仍有许多谜团。应对潜伏感染的新工具必须取代用于发展抗逆转录病毒治疗的范例和模式。现有的代表体内艾滋病毒潜伏期的细胞和动物模型需要进一步开发，虽然潜伏期前病毒可以在实验室中清除，但可测试的全面治疗策略尚不存在。因此，我们建议成立马丁·德莱尼合作实验室来根除艾滋病毒-1感染，这是21名杰出的研究人员的密切合作，他们在过去10年里共同领导了艾滋病毒潜伏期领域。为了最大限度地取得成功，我们将在四个研究领域开展工作，以开发定义根除疗法所需的基础设施和系统，识别具有治疗潜力的新分子，并在动物模型中为基于小分子的根除策略提供概念验证。目标1将确定病毒持续和潜伏的分子机制；目标2将确定减少潜伏病毒库的候选药物和治疗策略；目标3将建立信息丰富的动物模型系统以评估潜伏期和测试治疗策略；最后。目标4将在人类身上进行研究，以描绘病毒持续存在的基础。三个核心将在药理学、分子分析以及序列和表达分析方面协助研究项目。管理核心将确保协调，并保持这一经验丰富和强有力的团队对翻译产品开发的关注。作为一个团队，我们致力于集中我们的资源和专业知识，以超越学术研究的正常限制。值得注意的是，默克研究实验室的专业知识和持久承诺将是提供治疗进展的关键。我们相信，我们将共同推动取得进展，最终根除艾滋病毒感染。 公共卫生相关性：尽管抗逆转录病毒疗法(ART)在降低HLV-1感染患者死亡率方面取得了成功，但ART并未治愈这种疾病。接受有效抗逆转录病毒治疗的HIV患者T细胞中的持久病毒储备库是阻止HIV治愈的重要障碍。包括来自八所大学和默克研究实验室的科学家在内，马丁·德莱尼合作实验室将寻求通过开发和测试疗法来根除艾滋病毒感染，这些疗法最终能够在临床上进行测试，从而永久地摧毁病毒库。 项目1.1-确定调节艾滋病毒潜伏期的新宿主因素 项目负责人(PL)：华纳·格林 (申请人提供的描述)静态记忆CD4T细胞含有整合的、积极抑制的艾滋病毒前病毒，目前形成了根除病毒的强大障碍。通过激活前病毒基因的表达，从而使病毒对抗逆转录病毒治疗敏感，可以攻击这个潜伏的蓄水池。为了取得成功，这一策略必须既防止病毒传播，又导致所有潜伏感染的细胞死亡，而不会产生普遍的细胞激活的有毒状态。目前，还没有有效的治疗艾滋病毒潜伏期的方法。我们假设，更全面地了解HIV潜伏期的分子基础--尤其是促进和对抗潜伏期的宿主调控因素的全面范围--将有助于开发有效的治疗方法。在目标1中，我们将检查通过对感染艾滋病毒的HeLa细胞进行全基因组siRNA筛选而确定的五个新的候选艾滋病毒抑制因子。这些候选基因具有与潜伏期作用相一致的生物学特性，并在淋巴组织中表达。这些候选基因的表达将在生物相关的细胞中进行分析，并通过慢病毒shRNA敲除来评估它们的功能。在目标2中，我们将筛选HIV微小RNAs(MiRs)潜伏期的CD4T细胞模型，该模型通过损害细胞激活物的表达来促进病毒潜伏期。我们将在潜伏感染的原代CD4T细胞中使用抗配子来验证miR的作用。在目标3中，我们将使用生物信息学和转录图谱方法来鉴定被这些miRs抑制的宿主基因产物。使用这种双重实验方法，我们将确定自然促进和拮抗HIV潜伏期的细胞因素。在适当的情况下，将进行行动机制研究。将根据其活性的稳健性和总体的“可药性”，确定确定的目标的优先顺序，以便进入小分子高通量筛选试验，并将开发一次和二次筛选试验。这些拟议的研究可以识别出小分子，它们要么抑制最有趣的细胞抑制因子，要么激活最引人注目的受mRNA调控的艾滋病毒激活剂。这些研究完全支持合作实验室的目标--确定能够根除病毒或产生功能性治疗(无药物缓解)的无毒小分子组合。 与公共卫生相关：这些研究有望加深我们对促进和对抗艾滋病毒潜伏期的细胞因素的理解，并最终提供从潜伏库中清除艾滋病毒的新方法。最终，我们试图将这些对HIV潜伏期的基本见解转化为小分子诱导剂的组合，从而有效地清除潜伏期。这些研究的成功可能会从根本上改变世界各地艾滋病毒感染者的临床护理格局。

DESCRIPTION (provided by applicant): Despite the clinical success of antiretroviral therapy (ART), more people contract human immunodeficiency virus (HIV) infection daily than initiate ART. The difficulties of lifelong ART - particularly in the developing world - make the eradication of HIV imperative. But clearance of a retroviral infection for patients on ART is a herculean task. While much is known about HIV persistence despite ART, many puzzles remain. New tools to address latent infection must replace the paradigms and models used to develop ART. Existing cellular and animal models that represent HIV latency in vivo require further development, and while latent provirus can be purged in the laboratory, a testable, comprehensive therapeutic strategy is not at hand. Therefore we propose the Martin Delaney Collaboratory to Eradicate HIV-1 Infection, a close collaboration of 21 exceptional investigators who have collectively led the field of HIV latency over the last 10 years. To maximize success, we will work across four areas of research to develop the infrastructure and systems needed to define eradication therapies, identify new molecules with therapeutic potential and provide a proof-of-concept for a small molecule based eradication strategy in animal models. Objective 1 will identify the molecular mechanisms underlying viral persistence and latency; Objective 2 will identify drug candidates and therapeutic strategies to reduce the latent viral pool; Objective 3 will establish informative animal model systems to evaluate latency and test therapeutic strategies; and finally. Objective 4 will perform studies in humans to delineate the basis for viral persistence. Three cores will assist research projects with pharmacology, molecular assays, and sequence and expression analysis. An administrative core will assure coordination, and maintain the focus of this experienced and potent group towards translational product development. As a group, we are committed to pooling our resources and expertise to transcend the normal constraints of academic research. Of note, the expertise and durable commitment of Merck Research Laboratories will be critical to delivering therapeutic advances. We are convinced that together we will catalyze advances that will ultimately lead to the eradication of HIV infection. PUBLIC HEALTH RELEVANCE: Despite the success of antiretroviral therapy (ART) in decreasing mortality for HlV-1-infected patients, ART has not cured the disease. A persistent viral reservoir in the T cells of HIV patients receiving potent ART is a significant barrier preventing an HIV cure. Including scientists from eight universities and Merck Research Laboratories, the Martin Delaney Collaboratory will seek to eradicate HIV infection by developing and testing therapies, capable of eventually being tested clinically, that will permanently destroy the viral reservoir. Project 1.1 - Identification Novel Host Factors Regulating HIV Latency Project Leader (PL): Warner Greene (Description as provided by applicant) Quiescent memory CD4 T cells harboring integrated, actively repressed HIV proviruses currently form a formidable barrier to viral eradication. This latent reservoir could be attacked by activating proviral gene expression thereby sensitizing the virus to antiretroviral therapy. For success, this strategy must both prevent viral spread and result in the death of all latently infected cells without producing a toxic state of generalized cellular activation. Currently, no effective therapies for HIV latency exist. We hypothesize that a more complete understanding of the molecular underpinnings of HIV latency - notably, the full range of the host's regulatory factors that promote and antagonize latency - will facilitate the development of effective therapies. In Aim 1, we will examine five novel candidate HIV repressors identified by genome wide siRNA screening of HIV-infected HeLa cells. These candidates have biological properties consistent with a role in latency and are expressed in lymphoid tissues. Expression of these candidate genes will be analyzed in biologically relevant cells and their function assessed by lentiviral shRNA knockdown. In Aim 2, we will screen a CD4 T-cell model of HIV latency for microRNAs (miRs) that promotes viral latency by impairing the expression of cellular activators. We will validate miR action using antagomirs in latently infected primary CD4 T-cells. In Aim 3, we will use bioinformatic and transcriptional profiling approaches to identify the host gene products that are suppressed by these miRs. Using this dual experimental approach, we will identify cellular factors that naturally promote and antagonize HIV latency. Where appropriate, mechanism-of-action studies will be performed. Identified targets will be prioritized, based on the robustness of their activity and overall "drugability," for entry into small-molecule high-throughput screening assays and primary and secondary screening assays will be developed. These proposed studies could identify small molecules that either inhibit the most interesting cellular repressors or activate the most compelling mRNA-regulated HIV activators. These studies fully support the goal of the Collaboratory - to identify combinations of nontoxic small molecules that can eradicate the virus or produce a functional cure (drug-free remission). PUBLIC HEALTH RELEVANCE: These studies promise to deepen our understanding of the cellular factors that both promote and antagonize HIV latency and ultimately to provide new approaches for purging HIV from the latent reservoir. Ultimately, we seek to translate these basic insights into HIV latency into combinations of small molecule inducers that effectively purge the latent reservoir. Success in these studies could radically change the landscape of clinical care for HIV-infected patients throughout the world.