The Role of CBFb-MYH11 in Acute Myeloid Leukemia

影响核心结合因子（CBF）基因的易位是最常见的遗传畸变之一， 人类急性髓细胞白血病（AML），占所有AML的25%。其中近一半是 与影响16号染色体的易位相关，导致融合蛋白CBF的表达[3- MYH 11.拟议研究的总体目标是确定信号通路和下游 研究CBF（3-MYH 11）调控的靶基因，以评估这些调控基因在白血病发病中的作用。 干细胞自我更新，存活和分化，并确定与干细胞相互作用的信号通路。 CBFP-MYH 11在白血病发生中的作用为了实现这一目标，我们提出使用融合基因模型 我们已经开发了在人CD 34+细胞中的表达，以研究CBFP导致的白血病前期， MYH 11在获得导致明显白血病的协同突变之前的表达。我们有 利用该系统成功构建了相关白血病融合蛋白AML 1-ETO。的具体目标 目的1：明确CBF（3-MYH 11）下游信号通路。 CBF β-MYH 11在人CD 34+细胞中的表达导致其在体外持续生长， 超过五个月的期限。这些细胞的特征是它们的增殖，存活 和分化，并与相关CBF融合AML 1-ETO启动的长期培养进行比较。 目的2：鉴定CBFp-MYH 11的靶基因，并分析其在CBFp-MYH 11诱导的细胞增殖中的作用。 将通过比较CD 34+细胞的转录组来鉴定共同的基因表达谱， 我们的长期培养与正常的CD 34+细胞。我们将分析这些调控目标的贡献 通过过度表达那些被抑制的基因，反过来通过敲低那些成为 上调。目的3：确定与CBFp-MYH 11在白血病发生中协同作用的信号通路。 表达CBFp-MYH 11的白血病前培养物将作为背景，在其中合作 突变将被测试。识别将在急性转化中合作的特定信号 白血病、确定的遗传元件和饱和逆转录病毒突变将被使用。

Translocations affecting a core binding factor (CBF) gene are one of the most frequent genetic aberrations in human acute myeloid leukemia (AML), accounting for up to 25% of all AMLs. Nearly half of these are associated with translocations affecting chromosome 16, resulting in expression of the fusion protein CBF[3- MYH11. The overall objective of the proposed studies is to define the signaling pathways and downstream target genes that are affected by CBF(3-MYH11,to evaluate the role of these regulated genes in leukemic stem cell self-renewal, survival and differentiation,and to identify the signaling pathways that cooperate with CBFP-MYH11 in leukemogenesis. To accomplish this goal, we propose to use the model of fusion gene expression in human CD34+ cells we have developed, to study the pre-leukemia that is a result of CBFP- MYH11 expression prior to the acquisition of cooperating mutations that lead to frank leukemia. We have successfully used this system to model the related leukemia fusion protein AML1-ETO. The specific aims of this proposal are as follows: Aim 1: Define the signaling pathways downstream of CBF(3-MYH11. Expression of CBF(3-MYH11 in human CD34+ cells leads to their continued growth in vitro for an extended period of greater than five months. These cells will be characterizedin terms of their proliferation, survival and differentiation,and compared to the long-term cultures initiated by the related CBF fusion, AML1-ETO. Aim 2: Identify target genes of CBFp-MYHl1 and analyze their role in CBFp-MYHl 1 -inducedproliferation. A common gene expression profile will be identified by comparing the transcriptome of CD34+ cells from our long-term cultures with normal CD34+ cells. We will analyze the contribution of these regulated target genes by over-expressing those that are repressed, and conversely by knocking-down those genes that become upregulated. Aim 3: Identify the signaling pathways that cooperate with CBFp-MYHl1 in leukemogenesis. The pre-leukemic cultures expressing CBFp-MYHl1 will serve as the background in which cooperating mutations will be tested. To identify specific signals that will cooperate in the transformation to acute leukemia, defined eenetic elements and saturatine retroviral mutaeenesis will be used.