Single-Cell Analysis of HIV Latency Reversal

HIV潜伏期逆转的单细胞分析

• 批准号: 9203758
• 负责人: Jonathan Li
• 金额: $ 23.84万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-20 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9203758
• 关键词: Address; Biological Assay; CD4 Positive T Lymphocytes; Cell Count; Cell Proliferation; Cells; Chromosomes; Collaborations; DNA; Development; Disease remission; Effectiveness; Etiology; Evaluation; Exposure to; Frequencies; Genes; Genetic Transcription; HIV; HIV Genome; HIV Infections; Heterogeneity; Individual; Life; Location; Measures; Methods; Nature; Participant; Pathogenesis; Patients; RNA; Role; Sampling; Site; Study Section; Techniques; Technology; Testing; Therapeutic; Transcript; Viral; Work; antiretroviral therapy; cell fixing; cost; cost efficient; design; integration site; novel; novel strategies; response; single cell analysis; viral RNA

PROJECT SUMMARY The persistence of HIV within long-lived reservoirs results in the need for life-long antiretroviral therapy (ART). The accurate quantification of these reservoirs is challenging due to the relatively small number of latently infected cells within an individual. The optimal assay for HIV reservoir quantification should be high-throughput, sensitive, cost-efficient, and would ideally be able to quantify the frequency of HIV-infected cells. Current assays to measure the size of these reservoirs are hindered by a number of drawbacks, including the inability to determine infected cell numbers, the need for large sample volumes, and the cost and labor-intensive nature of certain assays. A single-cell assay that can quantify the number of infected cells with active HIV transcription would provide an unprecedented level of clarity on the state of the HIV reservoir, the effects of latency- reversing agents, and would have important implications for mechanistic and basic pathogenesis studies as well. We have recently validated a single-cell platform for detecting the frequency of HIV-expressing cells. Using this assay, we propose to evaluate the effectiveness of a range of latency-reversing agents for HIV reactivation and to explore a possible underlying mechanism behind their differential effects.

项目摘要 艾滋病毒在长寿命的储层中的持久性导致需要终身抗逆转录病毒疗法（ART）。 由于较少的潜在数量，这些储层的准确量化是具有挑战性的 一个人内感染细胞。 HIV储层定量的最佳测定法应该是高通量的， 敏感，成本效益，理想情况下可以量化感染HIV感染细胞的频率。当前的 测量这些储层大小的测定受到许多缺点的阻碍，包括无法 为了确定受感染的细胞数量，需要大量样品量以及成本和劳动密集型性质 某些测定法。单细胞测定，可以用活跃的HIV转录来量化感染细胞的数量 将为艾滋病毒水库的状态提供前所未有的清晰度，潜伏期的影响 逆转剂，并将对机械和基本发病机理研究具有重要意义 出色地。我们最近验证了一个单细胞平台，用于检测表达艾滋病毒细胞的频率。 使用此测定，我们建议评估一系列艾滋病毒的延迟逆转剂的有效性 重新激活并探索其差异作用背后的潜在机制。