The MN1 oncogene is deregulated in human acute myeloid leukemia and its overexpression induces proliferation and represses myeloid differentiation of primitive human and mouse hematopoietic cells, leading to myeloid leukemia in mouse models. To delineate the sequences within MN1 necessary for MN1-induced leukemia, we tested the transforming capacity of in-frame deletion mutants, using retroviral transduction of mouse bone marrow. We found that integrity of the regions between amino acids 12 to 458 and 1119 to 1273 are required for MN1’s in vivo transforming activity, generating myeloid leukemia with some mutants also producing T-cell lympho-leukemia and megakaryocytic leukemia. Although both full length MN1 and a mutant that lacks the residues between 12–228 (Δ12–228 mutant) repressed myeloid differentiation and increased myeloproliferative activity in vitro, the mutant lost its transforming activity in vivo. Both MN1 and Δ12–228 increased the frequency of common myeloid progentiors (CMP) in vitro and microarray comparisons of purified MN1-CMP and Δ12–228-CMP cells showed many differentially expressed genes including Hoxa9, Meis1, Myb, Runx2, Cebpa, Cebpb and Cebpd. This collection of immediate MN1-responsive candidate genes distinguishes the leukemic activity from the in vitro myeloproliferative capacity of this oncoprotein.
References
[1]
Rosenbauer F, Tenen DG (2007) Transcription factors in myeloid development: balancing differentiation with transformation. Nat Rev Immunol 7: 105–117.
Meester-Smoor MA, Vermeij M, van Helmond MJ, Molijn AC, van Wely KH, et al. (2005) Targeted disruption of the Mn1 oncogene results in severe defects in development of membranous bones of the cranial skeleton. Mol Cell Biol 25: 4229–4236.
[4]
Buijs A, Sherr S, van Baal S, van Bezouw S, van der Plas D, et al. (1995) Translocation (12;22) (p13;q11) in myeloproliferative disorders results in fusion of the ETS-like TEL gene on 12p13 to the MN1 gene on 22q11. Oncogene 10: 1511–1519.
[5]
Valk PJ, Verhaak RG, Beijen MA, Erpelinck CA, Barjesteh van Waalwijk van Doorn-Khosrovani S, et al. (2004) Prognostically useful gene-expression profiles in acute myeloid leukemia. N Engl J Med 350: 1617–1628.
[6]
Ross ME, Mahfouz R, Onciu M, Liu HC, Zhou X, et al. (2004) Gene expression profiling of pediatric acute myelogenous leukemia. Blood 104: 3679–3687.
[7]
Carella C, Bonten J, Sirma S, Kranenburg TA, Terranova S, et al. (2007) MN1 overexpression is an important step in the development of inv(16) AML. Leukemia 21: 1679–1690.
[8]
Baldus CD, Mrozek K, Marcucci G, Bloomfield CD (2007) Clinical outcome of de novo acute myeloid leukaemia patients with normal cytogenetics is affected by molecular genetic alterations: a concise review. Br J Haematol 137: 387–400.
[9]
Heuser M, Beutel G, Krauter J, Dohner K, von Neuhoff N, et al. (2006) High meningioma 1 (MN1) expression as a predictor for poor outcome in acute myeloid leukemia with normal cytogenetics. Blood 108: 3898–3905.
[10]
Slape C, Hartung H, Lin YW, Bies J, Wolff L, et al. (2007) Retroviral insertional mutagenesis identifies genes that collaborate with NUP98-HOXD13 during leukemic transformation. Cancer Res 67: 5148–5155.
[11]
Liu T, Jankovic D, Brault L, Ehret S, Baty F, et al. (2010) Functional characterization of high levels of meningioma 1 as collaborating oncogene in acute leukemia. Leukemia 24: 601–612.
[12]
Heuser M, Argiropoulos B, Kuchenbauer F, Yung E, Piper J, et al. (2007) MN1 overexpression induces acute myeloid leukemia in mice and predicts ATRA resistance in patients with AML. Blood 110: 1639–1647.
[13]
Kandilci A, Grosveld GC (2009) Reintroduction of CEBPA in MN1-overexpressing hematopoietic cells prevents their hyperproliferation and restores myeloid differentiation. Blood 114: 1596–1606.
[14]
Buijs A, van Rompaey L, Molijn AC, Davis JN, Vertegaal AC, et al. (2000) The MN1-TEL fusion protein, encoded by the translocation (12;22)(p13;q11) in myeloid leukemia, is a transcription factor with transforming activity. Mol Cell Biol 20: 9281–9293.
[15]
Boer J, Bonten-Surtel J, Grosveld G (1998) Overexpression of the nucleoporin CAN/NUP214 induces growth arrest, nucleocytoplasmic transport defects, and apoptosis. Mol Cell Biol 18: 1236–1247.
[16]
Akashi K, Traver D, Miyamoto T, Weissman IL (2000) A clonogenic common myeloid progenitor that gives rise to all myeloid lineages. Nature 404: 193–197.
[17]
Irizarry RA, Hobbs B, Collin F, Beazer-Barclay YD, Antonellis KJ, et al. (2003) Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 4: 249–264.
[18]
Huang da W, Sherman BT, Lempicki RA (2009) Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 4: 44–57.
[19]
Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, et al. (2005) Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci U S A 102: 15545–15550.
[20]
Cardone M, Kandilci A, Carella C, Nilsson JA, Brennan JA, et al. (2005) The novel ETS factor TEL2 cooperates with Myc in B lymphomagenesis. Mol Cell Biol 25: 2395–2405.
[21]
Carella C, Bonten J, Rehg J, Grosveld GC (2006) MN1-TEL, the product of the t(12;22) in human myeloid leukemia, immortalizes murine myeloid cells and causes myeloid malignancy in mice. Leukemia 20: 1582–1592.
[22]
Heuser M, Yun H, Berg T, Yung E, Argiropoulos B, et al. (2011) Cell of origin in AML: susceptibility to MN1-induced transformation is regulated by the MEIS1/AbdB-like HOX protein complex. Cancer cell 20: 39–52.
[23]
Hess JL, Bittner CB, Zeisig DT, Bach C, Fuchs U, et al. (2006) c-Myb is an essential downstream target for homeobox-mediated transformation of hematopoietic cells. Blood 108: 297–304.
[24]
Jin S, Zhao H, Yi Y, Nakata Y, Kalota A, et al. (2010) c-Myb binds MLL through menin in human leukemia cells and is an important driver of MLL-associated leukemogenesis. J Clin Invest 120: 593–606.
[25]
Slany RK (2009) The molecular biology of mixed lineage leukemia. Haematologica 94: 984–993.
[26]
Kuo YH, Zaidi SK, Gornostaeva S, Komori T, Stein GS, et al. (2009) Runx2 induces acute myeloid leukemia in cooperation with Cbfbeta-SMMHC in mice. Blood 113: 3323–3332.
[27]
Zhang P, Iwasaki-Arai J, Iwasaki H, Fenyus ML, Dayaram T, et al. (2004) Enhancement of hematopoietic stem cell repopulating capacity and self-renewal in the absence of the transcription factor C/EBP alpha. Immunity 21: 853–863.
[28]
Radomska HS, Basseres DS, Zheng R, Zhang P, Dayaram T, et al. (2006) Block of C/EBP alpha function by phosphorylation in acute myeloid leukemia with FLT3 activating mutations. The Journal of experimental medicine 203: 371–381.
[29]
Hirai H, Zhang P, Dayaram T, Hetherington CJ, Mizuno S, et al. (2006) C/EBPbeta is required for ‘emergency’ granulopoiesis. Nat Immunol 7: 732–739.
