The role of RUNX1 in MLL-AF4 leukaemia

RUNX1在MLL-AF4白血病中的作用

• 批准号: 1960359
• 金额: --
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1960359
• 关键词: role; RUNX1; MLL; AF4; leukaemia

During the past decades advances in our understanding of the biology of paediatric acute lymphoblastic leukaemias and optimization of therapeutic protocols have led to survival rates up to 85%. However, childhood leukaemias caused by translocation of MLL (Mixed Lineage Leukaemia) gene and its fusion with different partner genes are characterized by poor survival rates. Leukaemias caused by MLL-AF4 fusion protein are the most frequent among the MLL rearranged leukaemias and are characterized by small number of cooperating mutations and a very short latency, which underlines the role of MLL as a driver in the process of leukaemogenesis.The goal of this project will be to elucidate the role of specific transcription factors, previously identified in the lab, to MLL leukaemogenesis. The transcription factors involved in leukaemogenesis fall into two classes: (i) transcription factors that are downstream targets of the MLL-AF4 fusion protein; these are upregulated in MLL rearranged leukaemias and contribute directly to leukaemogenesis; and (ii) transcription factors that are normally expressed in lymphoid cells, but their normal function is abrogated in leukaemia cells due to the altered chromatin landscape and epigenetic signatures caused by the fusion protein.The binding patterns of the identified transcription factors will be investigated by high-throughput methods including ChIP-sequencing in the SEM leukaemia cell line, which harbours the MLL-AF4 mutation. In this way I will investigate whether this is affected after treatment with small molecule inhibitors that abrogate leukaemic growth. In addition, the role of the identified transcription factors will be examined by performing knockdown experiments in the SEM cell line, in order to determine the effects on leukaemic growth. Furthermore, I will employ newly developed techniques, including CITE-Seq and REAP-Seq, that combine high-throughput single cell RNA-sequencing with cell surface protein detection. This will enable us to examine the expression of different transcription factors in primary MLL-AF4 cells, compared to normal B cells and B cell progenitors, and will provide valuable insight into the origin of MLL leukaemias and the molecular biology of the disease.

在过去的几十年里，我们对儿科急性淋巴细胞白血病生物学的理解和治疗方案的优化使生存率高达85%。然而，由MLL（混合谱系白血病）基因易位及其与不同伴侣基因融合引起的儿童白血病的特征是生存率低。MLL-AF 4融合蛋白引起的白血病是MLL重排白血病中最常见的一种，其特点是协同突变数量少，潜伏期短，这突出了MLL在白血病发生过程中的驱动作用。参与白血病发生的转录因子分为两类：（i）作为MLL-AF 4融合蛋白下游靶点的转录因子;这些转录因子在MLL重排白血病中上调并直接促进白血病发生;和（ii）通常在淋巴细胞中表达的转录因子，但是由于融合蛋白引起的染色质景观和表观遗传特征的改变，它们的正常功能在白血病细胞中被废除。将通过高通量方法研究转录因子，包括在携带MLL-AF 4突变的SEM白血病细胞系中进行ChIP测序。通过这种方式，我将调查是否这是受小分子抑制剂治疗后，废除白血病的生长。此外，将通过在SEM细胞系中进行敲减实验来检查所鉴定的转录因子的作用，以确定对白血病生长的影响。此外，我将采用新开发的技术，包括CITE-Seq和REAP-Seq，这些技术将联合收割机高通量单细胞RNA测序与细胞表面蛋白检测相结合。这将使我们能够检查与正常B细胞和B细胞祖细胞相比，原代MLL-AF 4细胞中不同转录因子的表达，并将为MLL白血病的起源和该疾病的分子生物学提供有价值的见解。