Department of Hematology, Oncology, Hemostaseology, and Stem Cell Transplantation, Faculty of Medicine, RWTH Aachen University, Aachen, Germany.
Joint Research Center for Computational Biomedicine, RWTH Aachen University, Aachen, Germany.
Leukemia. 2020 Apr;34(4):1062-1074. doi: 10.1038/s41375-019-0629-z. Epub 2019 Nov 14.
Classical Philadelphia chromosome-negative myeloproliferative neoplasms (MPN) are a heterogeneous group of hematopoietic malignancies including polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). The JAK2V617F mutation plays a central role in these disorders and can be found in 90% of PV and ~50-60% of ET and PMF. Hypoxia-inducible factor 1 (HIF-1) is a master transcriptional regulator of the response to decreased oxygen levels. We demonstrate the impact of pharmacological inhibition and shRNA-mediated knockdown (KD) of HIF-1α in JAK2V617F-positive cells. Inhibition of HIF-1 binding to hypoxia response elements (HREs) with echinomycin, verified by ChIP, impaired growth and survival by inducing apoptosis and cell cycle arrest in Jak2V617F-positive 32D cells, but not Jak2WT controls. Echinomycin selectively abrogated clonogenic growth of JAK2V617F cells and decreased growth, survival, and colony formation of bone marrow and peripheral blood mononuclear cells and iPS cell-derived progenitor cells from JAK2V617F-positive patients, while cells from healthy donors were unaffected. We identified HIF-1 target genes involved in the Warburg effect as a possible underlying mechanism, with increased expression of Pdk1, Glut1, and others. That was underlined by transcriptome analysis of primary patient samples. Collectively, our data show that HIF-1 is a new potential therapeutic target in JAK2V617F-positive MPN.
经典费城染色体阴性骨髓增殖性肿瘤(MPN)是一组异质性造血恶性肿瘤,包括真性红细胞增多症(PV)、原发性血小板增多症(ET)和原发性骨髓纤维化(PMF)。JAK2V617F 突变在这些疾病中起着核心作用,可在 90%的 PV 和~50-60%的 ET 和 PMF 中发现。缺氧诱导因子 1(HIF-1)是对低氧水平反应的主要转录调节因子。我们研究了药理学抑制和 shRNA 介导的 HIF-1α 敲低(KD)对 JAK2V617F 阳性细胞的影响。用依曲膦抑制 HIF-1 与低氧反应元件(HREs)的结合,通过 ChIP 验证,在 Jak2V617F 阳性 32D 细胞中诱导凋亡和细胞周期停滞,从而损害生长和存活,但对 Jak2WT 对照没有影响。依曲膦选择性地消除了 JAK2V617F 细胞的集落形成能力,并降低了 JAK2V617F 阳性患者的骨髓和外周血单核细胞以及 iPS 细胞衍生祖细胞的生长、存活和集落形成,而来自健康供体的细胞不受影响。我们确定了参与瓦博格效应的 HIF-1 靶基因作为可能的潜在机制,包括 Pdk1、Glut1 等的表达增加。这一点通过对原发性患者样本的转录组分析得到了强调。总的来说,我们的数据表明 HIF-1 是 JAK2V617F 阳性 MPN 的一个新的潜在治疗靶点。