Müller-Tidow Carsten, Steffen Björn, Cauvet Thomas, Tickenbrock Lara, Ji Ping, Diederichs Sven, Sargin Bülent, Köhler Gabriele, Stelljes Matthias, Puccetti Elena, Ruthardt Martin, deVos Sven, Hiebert Scott W, Koeffler H Phillip, Berdel Wolfgang E, Serve Hubert
Department of Medicine, Gerhard Domagk Institute of Pathology, University of Münster, Münster, Germany.
Mol Cell Biol. 2004 Apr;24(7):2890-904. doi: 10.1128/MCB.24.7.2890-2904.2004.
The acute myeloid leukemia (AML)-associated translocation products AML1-ETO, PML-retinoic acid receptor alpha (RARalpha), and PLZF-RARalpha encode aberrant transcription factors. Several lines of evidence suggest similar pathogenetic mechanisms for these fusion proteins. We used high-density oligonucleotide arrays to identify shared target genes in inducibly transfected U937 cells expressing AML1-ETO, PML-RARalpha, or PLZF-RARalpha. All three fusion proteins significantly repressed the expression of 38 genes and induced the expression of 14 genes. Several of the regulated genes were associated with Wnt signaling. One of these, plakoglobin (gamma-catenin), was induced on the mRNA and protein level by all three fusion proteins. In addition, primary AML blasts carrying one of the fusion proteins significantly overexpressed plakoglobin. The plakoglobin promoter was cloned and shown to be induced by AML1-ETO, with promoter activation depending on the corepressor and histone deacetylase binding domains. The induction of plakoglobin by AML fusion proteins led to downstream signaling and transactivation of TCF- and LEF-dependent promoters, including the c-myc promoter, which was found to be bound by plakoglobin in vivo after AML1-ETO expression. beta-Catenin protein levels and TCF and LEF target genes such as c-myc and cyclin D1 were found to be induced by the fusion proteins. On the functional level, a dominant negative TCF inhibited colony growth of AML1-ETO-positive Kasumi cells, whereas plakoglobin transfection into myeloid 32D cells enhanced proliferation and clonal growth. Injection of plakoglobin-expressing 32D cells into syngeneic mice accelerated the development of leukemia. Transduction of plakoglobin into primitive murine hematopoietic progenitor cells preserved the immature phenotype during colony growth, suggesting enhanced self-renewal. These data provide evidence that activation of Wnt signaling is a common feature of several balanced translocations in AML.
急性髓系白血病(AML)相关的易位产物AML1-ETO、早幼粒细胞白血病视黄酸受体α(RARα)和早幼粒细胞白血病锌指蛋白-RARα编码异常转录因子。多条证据表明这些融合蛋白具有相似的致病机制。我们使用高密度寡核苷酸阵列来鉴定在可诱导转染的表达AML1-ETO、PML-RARα或PLZF-RARα的U937细胞中的共同靶基因。所有这三种融合蛋白均显著抑制38个基因的表达并诱导14个基因的表达。一些受调控的基因与Wnt信号通路相关。其中之一,桥粒斑蛋白(γ-连环蛋白),在mRNA和蛋白水平上均被这三种融合蛋白诱导。此外,携带其中一种融合蛋白的原发性AML原始细胞显著过表达桥粒斑蛋白。桥粒斑蛋白启动子被克隆,并显示受AML1-ETO诱导,启动子激活依赖于共抑制因子和组蛋白去乙酰化酶结合域。AML融合蛋白对桥粒斑蛋白的诱导导致下游信号传导以及TCF和LEF依赖启动子的反式激活,包括c-myc启动子,在AML1-ETO表达后在体内发现该启动子与桥粒斑蛋白结合。发现融合蛋白可诱导β-连环蛋白的蛋白水平以及TCF和LEF靶基因如c-myc和细胞周期蛋白D1。在功能水平上,显性负性TCF抑制AML1-ETO阳性Kasumi细胞的集落生长,而将桥粒斑蛋白转染到髓系32D细胞中可增强增殖和克隆生长。将表达桥粒斑蛋白的32D细胞注射到同基因小鼠中可加速白血病的发展。将桥粒斑蛋白转导到原始小鼠造血祖细胞中可在集落生长期间保留未成熟表型,提示自我更新增强。这些数据提供了证据表明Wnt信号通路的激活是AML中几种平衡易位的共同特征。