Chromatin and epigenetic landscape of differentiating CD34 haematopoietic cells and the impact of viral infection

造血是免疫系统正常发育的重要过程。驻留在骨髓中的CD 34+祖细胞通过尚不完全清楚的机制被定向为不同的细胞命运，但被假设为通过短距离和长距离表观遗传相互作用涉及基因表达的动态调节。CD 34+祖细胞是人巨细胞病毒（HCMV）终身潜伏感染的重要部位。有趣的是，HCMV优先在分化的髓样细胞中检测到，这表明病毒积极地调节细胞命运。我们的目的是了解在潜伏感染期间染色质和表观遗传学景观如何变化，并确定对髓样细胞定型至关重要的潜在特征。为了做到这一点，我们将使用荧光标记表达病毒来分类感染的细胞，并将它们培养7天，这项工作将在Reeves实验室（二级主管）完成，该实验室拥有丰富的HCMV专业知识，特别是对CD 34+细胞中病毒潜伏期的研究，并可使用UCL最先进的分选设备。一旦分离，我们将进行ChIP-seq以研究表观遗传标记（H3 K27 ac，H3 K4 me 1和H3 K4 me 3）和结构蛋白（CTCF和Cohesin）在人类基因组上的分布，并进行HiChIP以研究DNA在不同实验条件下的3D测序。我们还将提取RNA并对文库进行测序以检测基因表达的变化。这项工作将在Zabetlab（主要主管）完成，该实验室在染色质和表观遗传学方面具有广泛的专业知识，无论是wetlab还是计算。

Haematopoiesis is an essential process that underpins the normal development of ourimmune system. CD34+ progenitor cells resident in the bone marrow are committed todifferent cell fates through mechanisms that are not completely understood but arehypothesized to involve dynamic regulation of gene expression through short and long rangeepigenetic interactions. CD34+ progenitor cells are an important site for humancytomegalovirus (HCMV) lifelong latent infections in the host. Intriguingly, HCMV ispreferentially detected in differentiated myeloid cells suggesting that the virus activelyregulates cell fate.Our aim is to understand how does the chromatin and epigenetics landscapes change duringthe latent infection and to identify potential signatures crucial for myeloid cell commitment.To do this, we will use fluorescently tag expressing viruses to sort infected cells and culturethem for seven days to compare them with uninfected and bystander cells from the infection.This work will be done in the Reeves lab (the secondary supervisor) that has extensiveexpertise with HCMV, particularly studies of viral latency in CD34+ cells, and access to stateof-the-art sorting facilities at UCL. Once isolated, we will perform ChIP-seq to investigatedistribution of epigenetic marks (H3K27ac, H3K4me1 and H3K4me3) and of the architecturalproteins (CTCF and Cohesin) on the human genome and HiChIP to investigate the 3D wiringof the DNA under the different experimental conditions. We will also extract the RNA andsequence the libraries to detect changes in gene expression. This work will be done in Zabetlab (primary supervisor) that has extensive expertise in chromatin and epigenetics bothwetlab and computational.