Cooperating genes in inv(16) acute myeloid leukemia

描述（由申请人提供）：急性髓性白血病（AML）由造血祖细胞不受控制的克隆扩增引起。不同亚型的AML与特定的染色体易位相关。例如，M4Eo亚型与编码融合基因CBFb-MYH11的基因的16号染色体断裂-连接反转有关。在亚型中，在其他基因中也发现了额外的突变，包括Ras、p53和NFl。这些突变可能在CBFB-MYH11介导的AML中发挥作用。我们生成了Cbfb-MYH11敲入小鼠，模拟了Cbfb-MYH11在人类AML-M4Eo中的存在。我们已经证明Cbfb-MYH11通过阻断造血分化在白血病发生中发挥作用。我们假设AML是一个涉及两个事件的过程的结果：1。2.改变造血干细胞分化的融合基因的产生。引入一种或多种与细胞凋亡和/或增殖有关的其他突变。为了验证这些假设，我们将鉴定与Cbfb-MYH11协同发展AML的基因。我们将首先将我们敲入小鼠的逆转录病毒插入突变与反向PCR相结合，以鉴定AML中改变的基因（目的1）。然后，我们将利用已鉴定基因的逆转录病毒转导和移植实验来测试这些基因的转化作用。作为检验我们假设的另一种方法，我们将使用遗传杂交和逆转录病毒转导实验来评估Ras相关基因和Cbfb-MYH11在AML发展中的功能相互作用（目的2）。我们的长期目标是了解决定AML的遗传机制，并为改进疗法的设计提供靶点。

DESCRIPTION (provided by applicant): Acute myeloid leukemias (AML) arise from the uncontrolled clonal expansion of hematopoietic progenitor cells. Different subtypes of AML are associated with specific chromosomal translocations. For example, the subtype M4Eo is associated with the chromosome 16 break-and-join inversion of the genes that code for the fusion gene CBFb-MYH11. Within subtypes, additional mutations have also been found in other genes including Ras, p53, and NFl. These mutations could play a role in CBFB-MYH11 mediated AML .We generated the Cbfb-MYH11 knock-in mouse, mimicking the presence of CBFb-MYH11 in human AML-M4Eo. We have shown that Cbfb-MYH11 plays a role in leukemogenesis by blocking hematopoietic differentiation. We hypothesize that AML is the result of a process that involves two-events: 1. The creation of a fusion gene that alters hematopoietic stem cell differentiation and 2. the introduction of one or more other mutations that are associated with apoptosis and/or proliferation. To test these hypotheses we will identify genes that synergize with Cbfb-MYH11 to develop AML. We will first combine retroviral insertional mutagenesis in our knock in mice with inverse PCR to identify genes altered in AML (Aim 1). We will then use retroviral transduction of identified genes and transplantation experiments to test for a transforming role of these genes. As an alternative approach to test our hypotheses, we will use genetic crosses and retroviral transduction experiments to evaluate the functional interactions between Ras associated genes and Cbfb-MYH11 in AML development (Aim 2). Our long-term goals are to understand the genetic mechanisms that determine AML, and to provide targets for the design of improved therapies.