A novel signalling screen demonstrates that CALR mutations activate essential MAPK signalling and facilitate megakaryocyte differentiation.
A novel signalling screen demonstrates that CALR mutations activate essential MAPK signalling and facilitate megakaryocyte differentiation.
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DOI：
10.1038/leu.2016.280
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发表时间：
2017-04
期刊：
影响因子：
11.4
通讯作者：
Green T
中科院分区：
文献类型：
--
作者：
Kollmann K;Warsch W;Gonzalez-Arias C;Nice FL;Avezov E;Milburn J;Li J;Dimitropoulou D;Biddie S;Wang M;Poynton E;Colzani M;Tijssen MR;Anand S;McDermott U;Huntly B;Green T
大多数缺乏JAK2突变的骨髓增生性肿瘤（MPN）患者携带体细胞CALR突变，这些突变被认为可以激活细胞因子信号传导，尽管其机制尚不清楚。为了鉴定对calr突变细胞存活重要的激酶，我们开发了一种新的策略（KISMET），该策略利用了每种抑制剂提供的所有激酶选择性数据，从而利用了脱靶噪声，限制了传统的小干扰RNA或抑制剂筛选。KISMET成功鉴定了造血和非造血细胞系中已知的必需激酶，并鉴定了calr突变的MARIMO细胞生长所需的丝裂原激活蛋白激酶（MAPK）途径。在小鼠或人造血细胞系中，突变型CALR的表达伴随着骨髓增殖性白血病蛋白（MPL）依赖的MAPK信号激活，CALR突变的MPN患者在CD34细胞、血小板和巨核细胞中显示出增加的MAPK活性。尽管CALR突变导致蛋白质不稳定和蛋白体降解，但突变的CALR能够增强人类CD34+祖细胞的巨核生成和促血小板生成。这些数据将异常的MAPK激活与MPN表型联系起来，并将其确定为calr突变阳性MPN的潜在治疗靶点。
Most myeloproliferative neoplasm (MPN) patients lacking JAK2 mutations harbour somatic CALR mutations that are thought to activate cytokine signalling although the mechanism is unclear. To identify kinases important for survival of CALR-mutant cells, we developed a novel strategy (KISMET) that utilizes the full range of kinase selectivity data available from each inhibitor and thus takes advantage of off-target noise that limits conventional small-interfering RNA or inhibitor screens. KISMET successfully identified known essential kinases in haematopoietic and non-haematopoietic cell lines and identified the mitogen activated protein kinase (MAPK) pathway as required for growth of the CALR-mutated MARIMO cells. Expression of mutant CALR in murine or human haematopoietic cell lines was accompanied by myeloproliferative leukemia protein (MPL)-dependent activation of MAPK signalling, and MPN patients with CALR mutations showed increased MAPK activity in CD34 cells, platelets and megakaryocytes. Although CALR mutations resulted in protein instability and proteosomal degradation, mutant CALR was able to enhance megakaryopoiesis and pro-platelet production from human CD34+ progenitors. These data link aberrant MAPK activation to the MPN phenotype and identify it as a potential therapeutic target in CALR-mutant positive MPNs.