C-KIT mutation cooperates with full-length AML1-ETO to induce acute myeloid leukemia in mice
C-KIT mutation cooperates with full-length AML1-ETO to induce acute myeloid leukemia in mice
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DOI：
10.1073/pnas.1019625108
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发表时间：
2011-02-08
影响因子：
11.1
通讯作者：
Chen, Sai-Juan
中科院分区：
文献类型：
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作者：
Wang, Yue-Ying;Zhao, Li-Juan;Chen, Sai-Juan
人急性髓性白血病（AML）中由t（8;21）（q22;q22）产生的全长AML 1-ETO（AE）融合基因不足以在动物中诱导白血病，这表明白血病发生需要额外的突变。我们和其他人已经在近一半的t（8;21）AML患者中发现了C-KIT的激活突变。为了检验活化C-KIT突变与AE协同导致明显AML的假设，我们产生了具有突变C-KIT和AE两者的鼠转导和移植模型。为了克服人C-KIT在鼠细胞中的胞内转运阻断，我们通过将鼠c-Kit的胞外和跨膜结构域框内融合到人C-KIT的胞内信号传导结构域来工程化杂合C-KIT（HyC-KIT）。我们发现酪氨酸激酶结构域突变体HyC-KIT N822 K和D816 V以及跨膜突变体HyC-KIT 571+14和557- 558 Del可以将小鼠32 D细胞转化为非依赖于酪氨酸的生长。蛋白酪氨酸激酶抑制剂达沙替尼抑制表达这些C-KIT突变体的32 D细胞的增殖，效力在低纳摩尔范围内。在小鼠中，HyC-KIT N822 K诱导骨髓增生性疾病，而HyC-KIT 571+ 14诱导骨髓增生性疾病和淋巴细胞白血病。有趣的是，AE和HyC-KIT N822 K的共表达导致致命性AML。我们的数据进一步丰富了两次打击模型，转录因子和膜/胞质信号分子的异常是AML发病机制所必需的。此外，达沙替尼延长了携带AE和HyC-KIT N822 K共表达白血病细胞的小鼠的寿命，并与阿糖胞苷联合使用时发挥协同作用，从而为t（8;21）白血病提供了潜在的治疗方法。
The full-length AML1-ETO (AE) fusion gene resulting from t(8;21) (q22;q22) in human acute myeloid leukemia (AML) is not sufficient to induce leukemia in animals, suggesting that additional mutations are required for leukemogenesis. We and others have identified activating mutations of C-KIT in nearly half of patients with t (8;21) AML. To test the hypothesis that activating C-KIT mutations cooperate with AE to cause overt AML, we generated a murine transduction and transplantation model with both mutated C-KIT and AE. To overcome the intracellular transport block of human C-KIT in murine cells, we engineered hybrid C-KIT (HyC-KIT) by fusing the extracellular and transmembrane domains of the murine c-Kit in-frame to the intracellular signaling domain of human C-KIT. We showed that tyrosine kinase domain mutants HyC-KIT N822K and D816V, as well as juxtamembrane mutants HyC-KIT 571+14 and 557-558Del, could transform murine 32D cells to cytokine-independent growth. The protein tyrosine kinase inhibitor dasatinib inhibited the proliferation of 32D cells expressing these C-KIT mutants, with potency in the low nanomolar range. In mice, HyC-KIT N822K induced a myeloproliferative disease, whereas HyC-KIT 571+ 14 induces both myeloproliferative disease and lymphocytic leukemia. Interestingly, coexpression of AE and HyC-KIT N822K led to fatal AML. Our data have further enriched the two-hit model that abnormalities of both transcription factor and membrane/cytosolic signaling molecule are required in AML pathogenesis. Furthermore, dasatinib prolonged lifespan of mice bearing AE and HyC-KIT N822K-coexpressing leukemic cells and exerted synergic effects while combined with cytarabine, thus providing a potential therapeutic for t(8;21) leukemia.