Role of AML1/Runx1 isoforms in development, self-renewal and differentiation of haematopoietic progenitor/stem cells

这项研究计划的主要焦点是一种名为AML1/RUNX1的基因对血液干细胞的发育和调控。干细胞是当前生物学研究的主要焦点之一，因为干细胞具有自我复制和分化为多种成熟细胞的能力。AML1/RUNX1对成人血液系统的发育至关重要，并经常参与人类白血病的发生。AML1/RUNX1基因产生不同形式的蛋白质，可以竞争影响其他参与干细胞发育和功能的基因，以及白血病的发生。我们已经产生了基因改变的胚胎干细胞和小鼠实验系统，允许人工调节AML1/RUNX1基因。AML1/RUNX1基因的调控是通过现代双组分四环素诱导系统实现的。该系统的设计能够微调细胞中AML1/RUNX1蛋白的水平。这些AML1/RUNX1蛋白的影响将在培养的ES细胞、发育中的小鼠胚胎和成年小鼠中进行研究。另外两只带有由AML1/RUNX1控制的荧光标记基因的转基因小鼠也将被用于这项研究，以实现对表达AML1蛋白的细胞的准确识别。更好地理解干细胞调节和AML1在这一过程中的作用将对制定实用的再生医学方案具有潜在的意义。

The main focus of this research proposal is in the development and regulation of blood stem cells by a gene called AML1/Runx1. Stem cells are one of the main foci of current biology due to their potential to self-replicate and give rise to a variety of mature cells. AML1/Runx1 is critically important for the development of the adult blood system and is frequently involved in the development of human leukaemias. AML1/Runx1 gene produces different forms of the protein which can compete for the influence on other genes which are involved in stem cell development and function as well in the development of leukaemias. We have generated genetically altered embryonic stem (ES) cells and mouse experimental systems which allow AML1/Runx1 gene to be artificially regulated. Regulation of AML1/Runx1 gene is achieved by the use of a modern bi-component tetracycline inducible system. The design of the system enables fine tuning of levels of the AML1/Runx1 protein in cells. The influence of these AML1/Runx1 proteins will be studied in cultured ES cells as well as in developing mouse embryos and in the adult mouse. Two additional genetically modified mice which have fluorescent tagged genes controlled by AML1/Runx1 will also be used in this research to enable accurate identification of cells which express AML1 protein. Better understanding of stem cell regulation and the role of AML1 in this process will have potential implications for the development of protocols for practical regenerative medicine.