Regulation of HIV Latency by Host Cell Transcriptional and Epigenetic Networks

项目概要： 潜伏感染细胞的长寿储库的存在被认为是主要的 治疗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。这一领域的一个关键问题是， 已知潜伏感染细胞的性质，以及 调节延迟。我们最近发现延迟与不同的主机有关 细胞转录签名。我们的假设是这个信号代表了一个宿主细胞 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组实验中，我们将直接测试这个假设， 研究宿主细胞环境调节的分子机制 HIV转录的沉默。这将通过直接测试个人的角色来实现 从HIV转录的潜伏标记中提取基因。我们还将进行详细的 分析在潜伏感染细胞中起作用的表观遗传途径，并直接检查 它们对整合前病毒的影响。最后，我们将使用尖端技术， 结合延时显微镜和scRNAseq，发现细胞的转录组学特征 与潜伏期逆转相关。因此，该数据集将导致新的目标， 治疗操纵潜伏期，并指导进一步的实验，以测试这些假设， 体外和体内潜伏期模型。

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.