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

造血是维持机体免疫系统正常发育的重要过程。驻留在骨髓中的CD34+祖细胞通过尚未完全了解的机制决定不同的细胞命运，但假设涉及通过短程和长程表观遗传相互作用来动态调节基因表达。CD34+祖细胞是人类巨细胞病毒(HCMV)终生潜伏感染宿主的重要部位。有趣的是，HCMV优先在分化的髓系细胞中被检测到，这表明病毒积极地调节细胞的命运。我们的目的是了解在潜伏感染过程中染色质和表观遗传学环境是如何变化的，并识别对髓系细胞承诺至关重要的潜在特征。为此，我们将使用荧光标签表达病毒对感染细胞进行分类，并培养它们七天，将它们与感染的未感染细胞和旁观者细胞进行比较。这项工作将在里夫斯实验室进行，该实验室在HCMV方面拥有丰富的专业知识，特别是CD34+细胞中的病毒潜伏期的研究，并访问UCL最先进的分选设施。一旦分离，我们将进行芯片序列研究人类基因组和HiChIP上表观遗传标记(H3K27ac、H3K4me1和H3K4me3)和结构蛋白(CTCF和Coherin)的分布，以研究不同实验条件下DNA的3D连接。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(首席主管)完成，该实验室在染色质和表观遗传学方面拥有广泛的专业知识，包括实验室和计算。

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.