High Throughput Screening to Identify Antagonists of HIV-1 Latency

高通量筛选以鉴定 HIV-1 潜伏期拮抗剂

• 批准号: 7163959
• 负责人: JOSEPH DOUGHERTY
• 金额: $ 53.88万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7163959
• 关键词: clinical research; high throughput technology; infection; latent virus infection; library; model; small molecule

DESCRIPTION (provided by applicant): Project Summary: Highly active antiretroviral therapy (HAART) has had a fundamental impact on the control of HIV-1 infection. HAART is remarkably effective at suppressing HIV-1 replication; however, it cannot completely eradicate the virus even during long-term therapy. A primary cause for the lack of complete viral clearance is a population of latently infected cells that appear to be a source for viral rebound upon withdrawal of antiretroviral therapy, thus perpetuating the infection and replenishing the latent reservoirs. Attempts to eliminate the latently infected cells by activating them with cytokines and lymphokines have not met with success probably because this treatment is unable to reach all of the sites that harbor latent virus and carries with it a high degree of toxicity. A promising alternative method using small molecules with pharmacological properties that would penetrate even the most inaccessible viral reservoirs and subsequently activate latent HIV-1 proviruses could very well improve the prospects for clearance of HIV-1. In this scenario, antiretroviral drugs, virus-induced cytotoxicity and immune-mediated elimination would control infection of freshly activated cells, while coincident administration of the anti- latency drug would not only force the emergence of latent virus but would also prevent the establishment of new latency. We developed a latency model that can be used for high throughput screening (HTS). With this system a very reliable signal was detected in 96- and 384-well formats as characterized by excellent Z' scores ranging between 0.55 and 0.80. Here we propose to utilize our model system for HTS to identify small molecule antagonists of latency. Once our library of low molecular weight molecules has been screened, the most active compounds will be further characterized both to probe the mechanism underlying the maintenance and reactivation of latent virus as well as providing molecules for further therapeutic development. We believe this system provides an efficient, rapid and safe method to discover novel agents ultimately aimed at elimination of HIV-1 infection. Relevance: When a person is infected with HIV, a small percentage of the cells that have been infected harbor viruses that can remain dormant for long periods of time, a phenomenon known as viral latency. Existing antiviral therapies are able to control actively replicating virus, but they cannot purge the hidden, latent virus, so patients remain perpetually infected because the latent virus may be activated at any time, leading to new rounds of viral production. Our goal is to identify new compounds that help to understand how to flush out the dormant virus and eliminate the infection in order to aid in reversing the course of the HIV pandemic.

描述（由申请人提供）：项目摘要：高度活跃的抗逆转录病毒疗法（HAART）对HIV-1感染的控制产生了根本的影响。 Haart在抑制HIV-1复制方面非常有效。但是，即使在长期治疗期间，它也无法完全消除病毒。缺乏完全病毒清除率的主要原因是戒断抗逆转录病毒疗法后似乎是病毒反弹的潜在细胞的种群，从而使感染永存并补充潜在储层。试图通过用细胞因子和淋巴细胞激活潜在感染的细胞来消除它们，这可能没有取得成功，这可能是因为这种治疗方法无法到达含有潜在病毒的所有部位，并且具有高度的毒性。使用具有药理学特性的小分子的一种有希望的替代方法，即使是最无法接近的病毒储存剂，并随后激活潜在的HIV-1病毒，可以很好地改善清除HIV-1的前景。在这种情况下，抗逆转录病毒药物，病毒诱导的细胞毒性和免疫介导的消除将控制新鲜活化的细胞的感染，而抗潜伏药的同时给药不仅会迫使潜在病毒的出现，而且还将防止建立新的潜伏期。我们开发了一个可用于高吞吐量筛查（HTS）的延迟模型。使用该系统，以96和384孔格式检测到非常可靠的信号，其特征在于出色的Z'分数在0.55至0.80之间。在这里，我们建议利用我们的模型系统来识别潜伏期的小分子拮抗剂。一旦我们筛选了低分子量分子的库，将进一步表征最活跃的化合物，以探测潜在病毒的维持和重新激活的基础机制，并为进一步的治疗发育提供分子。我们认为，该系统提供了一种有效，快速和安全的方法，可以发现最终旨在消除HIV-1感染的新型药物。相关性：当一个人感染艾滋病毒时，一小部分被感染的港口病毒可能会长时间保持休眠状态，这是一种称为病毒潜伏期的现象。现有的抗病毒疗法能够控制主动复制病毒，但不能清除隐藏的潜在病毒，因此患者仍然会永久感染，因为可以随时激活潜在病毒，从而导致新的病毒产生。我们的目标是确定有助于了解如何冲洗休眠病毒并消除感染的新化合物，以帮助逆转HIV大流行病学。