C-KIT mutation cooperates with full-length AML1-ETO to induce acute myeloid leukemia in mice

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 108; no. 6; pp. 2450 - 2455
• Main Authors: Wang, Yue-Ying, Zhao, Li-Juan, Wu, Chuan-Feng, Liu, Ping, Shi, Lin, Liang, Yang, Xiong, Shu-Min, Mi, Jian-Qing, Chen, Zhu, Ren, Ruibao, Chen, Sai-Juan
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 08.02.2011; National Acad Sciences
• Subjects: 3T3 cells; Acute myeloid leukemia; Animal models; Animals; Antimetabolites, Antineoplastic - pharmacology; Biological Sciences; Blood; c-Kit protein; Cell growth; Cell proliferation; Cell Transformation, Neoplastic - genetics; Cell Transformation, Neoplastic - metabolism; Cell Transformation, Neoplastic - pathology; Cells; Chromosomes, Human, Pair 21 - genetics; Chromosomes, Human, Pair 21 - metabolism; Chromosomes, Human, Pair 8 - genetics; Chromosomes, Human, Pair 8 - metabolism; Core Binding Factor Alpha 2 Subunit - genetics; Core Binding Factor Alpha 2 Subunit - metabolism; cytarabine; Cytarabine - pharmacology; Data processing; Fusion protein; gene fusion; Genes; Genetic mutation; Humans; Hybrids; Intracellular signalling; Kinases; Leukemia; Leukemia, Lymphoid - genetics; Leukemia, Lymphoid - metabolism; Leukemia, Lymphoid - pathology; Leukemia, Myeloid, Acute - genetics; Leukemia, Myeloid, Acute - metabolism; Leukemia, Myeloid, Acute - pathology; Leukemogenesis; Life span; longevity; Lymphatic leukemia; lymphocytic leukemia; Mice; Mice, Transgenic; Molecular modelling; Molecules; mutants; Mutation; Myeloid leukemia; Myeloproliferative diseases; NIH 3T3 Cells; Oncogene Proteins, Fusion - genetics; Oncogene Proteins, Fusion - metabolism; pathogenesis; patients; Protein Structure, Tertiary; Protein-tyrosine kinase; Proto-Oncogene Proteins c-kit - genetics; Proto-Oncogene Proteins c-kit - metabolism; Receptors; Rodents; RUNX1 Translocation Partner 1 Protein; Signal transduction; synergism; Transcription factors; Translocation, Genetic - genetics; Transmembrane domains; Transplantation; tyrosine
• ISSN: 0027-8424; 1091-6490; 1091-6490
• DOI: 10.1073/pnas.1019625108

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.