Regulation of HIV Latency by Host Cell Transcriptional and Epigenetic Networks

项目总结： 潜伏感染细胞的长期存活库的存在是公认的主要 这是治愈HIV-1感染的障碍。因此，治疗策略的发展 消除这些水库是一个关键的科学目标。这个领域的一个关键问题是，几乎没有 已知潜伏感染细胞的性质，以及 规范潜伏期。我们最近发现，延迟与不同的主机相关 细胞转录签名。我们的假设是这个信号代表一个宿主细胞 转录和表观遗传程序，调节建立或维持 延迟。在我们这里提出的一系列实验中，我们将直接检验这一假设，并且 研究宿主细胞环境调节的分子机制 HIV转录沉默。这将通过直接测试个人的角色来实现 来自HIV转录中潜伏期签名的基因。我们还将进行详细的 分析在潜伏感染细胞中运行的表观遗传途径，并直接检查 它们对整合的前病毒的影响。最后，我们将使用尖端技术， 结合时间推移显微镜和scRNAseq来发现细胞的转录特征 这与潜伏期逆转有关。因此，此数据集将导致新的目标 潜伏期的治疗操作，并指导进一步的实验来验证这些假说 潜伏期的体外和体内模型。

Project summary: The existence of long-lived reservoirs of latently infected cells is recognized as a primary barrier to a cure for HIV-1 infection. Thus, the development of therapeutic strategies to eliminate these reservoirs is a critical scientific goal. A key problem in this field is that little is known about the nature of latently infected cells, and the molecular mechanisms that regulate latency. We have recently discovered that latency is associated with a distinct host cell transcriptional signature. Our hypothesis is that this signature represents a host cell transcriptional and epigenetic program that regulates establishment or maintenance of latency. In the set of experiments we propose here, we will test this hypothesis directly, and investigate the molecular mechanisms by which the host cell environment regulates silencing of HIV transcription. This will be achieved directly testing the roles of individual genes from the latency signature in HIV transcription. We will also undertake a detailed analysis of epigenetic pathways that operate in latently infected cells, and directly examine their impact on integrated proviruses. Finally, we will use cutting edge technology that combines time-lapse microscopy and scRNAseq to discover transcriptomic features of cells that correlate with latency reversal. Thus, this dataset will lead to new targets for therapeutic manipulation of latency and guide further experiment to test these hypotheses in in vitro and in vivo models of latency.