AML1, the Target of Multiple Chromosomal Translocations in Human Leukemia, Is Essential for Normal Fetal Liver Hematopoiesis

The AML1-CBF beta transcription factor is the most frequent target of chromosomal rearrangements in human leukemia. To investigate its normal function, we generated mice lacking AML1. Embryos with homozygous mutations in AML1 showed normal morphogenesis and yolk sac-derived erythropoiesis, but lacked fetal liver hematopoiesis and died around E12.5. Sequentially targeted AML1-/-es cell retained their capacity to differentiate into primitive erythroid cells in vitro; however, no myeloid or erythroid progenitors of definitive hematopoietic origin were detected in either the yolk sac or fetal livers of mutant embryos. Moreover, this hematopoietic defect was intrinsic to the stem cells in that AML1-/-ES cells failed to contribute to hematopoiesis in chimeric animals. These results suggest that AML1-regulated target genes are essential for definitive hematopoiesis of all lineages.

[1]  F. Collins,et al.  Fusion between transcription factor CBF beta/PEBP2 beta and a myosin heavy chain in acute myeloid leukemia. , 1993, Science.

[2]  Y. Yazaki,et al.  Generation of the AML1‐EVI‐1 fusion gene in the t(3;21)(q26;q22) causes blastic crisis in chronic myelocytic leukemia. , 1994, The EMBO journal.

[3]  C. Wijmenga,et al.  Molecular pathogenesis of the chromosome 16 inversion in the M4Eo subtype of acute myeloid leukemia. , 1995, Blood.

[4]  E. Wieschaus,et al.  Dosage requirements for runt in the segmentation of Drosophila embryos , 1986, Cell.

[5]  J. Ihle.  Interleukin-3 and Hematopoiesis (Part 1 of 2) , 1992 .

[6]  S. Burden,et al.  AML1 is expressed in skeletal muscle and is regulated by innervation , 1994, Molecular and cellular biology.

[7]  Y. Ito,et al.  PEBP2/PEA2 represents a family of transcription factors homologous to the products of the Drosophila runt gene and the human AML1 gene. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[8]  R. Lovell-Badge,et al.  Modulation of gene activity by consecutive gene targeting of one creatine kinase M allele in mouse embryonic stem cells. , 1991, Nucleic acids research.

[9]  A. Emery,et al.  Genetic variability , 1980, Nature.

[10]  P. Gruss,et al.  Multiple point mutations affecting the simian virus 40 enhancer. , 1983, Science.

[11]  I. Bar-Am,et al.  AML1, AML2, and AML3, the human members of the runt domain gene-family: cDNA structure, expression, and chromosomal localization. , 1994, Genomics.

[12]  Stuart H. Orkin,et al.  Transcription Factors and Hematopoietic Development (*) , 1995, The Journal of Biological Chemistry.

[13]  J. Lenz,et al.  Relative importance of elements within the SL3-3 virus enhancer for T-cell specificity , 1990, Journal of virology.

[14]  C. A. Thomas,et al.  Molecular cloning. , 1977, Advances in pathobiology.

[15]  D C Ward,et al.  Fusion of the TEL gene on 12p13 to the AML1 gene on 21q22 in acute lymphoblastic leukemia. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[16]  R. Hromas,et al.  Transcriptional regulation of interleukin 3 gene expression in T lymphocytes. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[17]  Y. Ito,et al.  Purification of a mouse nuclear factor that binds to both the A and B cores of the polyomavirus enhancer , 1990, Journal of virology.

[18]  S. Shurtleff,et al.  Heterogeneity in CBF beta/MYH11 fusion messages encoded by the inv(16)(p13q22) and the t(16;16)(p13;q22) in acute myelogenous leukemia. , 1995, Blood.

[19]  R. Weinberg,et al.  Extensive contribution of Rb‐deficient cells to adult chimeric mice with limited histopathological consequences. , 1994, The EMBO journal.

[20]  F. Collins,et al.  Overexpression of core-binding factor alpha (CBF alpha) reverses cellular transformation by the CBF beta-smooth muscle myosin heavy chain chimeric oncoprotein , 1995, Molecular and cellular biology.

[21]  J. Lenz,et al.  Identification of the SL3-3 virus enhancer core as a T-lymphoma cell-specific element , 1989, Journal of virology.

[22]  S. Shurtleff,et al.  TEL/AML1 fusion resulting from a cryptic t(12;21) is the most common genetic lesion in pediatric ALL and defines a subgroup of patients with an excellent prognosis. , 1995, Leukemia.

[23]  S Meyers,et al.  PEBP2/CBF, the murine homolog of the human myeloid AML1 and PEBP2 beta/CBF beta proto-oncoproteins, regulates the murine myeloperoxidase and neutrophil elastase genes in immature myeloid cells , 1994, Molecular and cellular biology.

[24]  P. Landrigan.  Childhood leukemias. , 1995, The New England journal of medicine.

[25]  M. Ohki,et al.  The Runt domain identifies a new family of heteromeric transcriptional regulators. , 1993, Trends in genetics : TIG.

[26]  J. Ihle,et al.  Interleukin-3 and hematopoiesis. , 1992, Chemical immunology.

[27]  B. Wieringa,et al.  Genetic variability of the murine creatine kinase B gene locus and related pseudogenes in different inbred strains of mice. , 1992, Genomics.

[28]  J. Rowley,et al.  AML1 and the 8;21 and 3;21 translocations in acute and chronic myeloid leukemia. , 1995, Blood.

[29]  N. Speck,et al.  Cooperative binding of Ets-1 and core binding factor to DNA , 1994, Molecular and cellular biology.

[30]  E. Soeda,et al.  Alternative splicing and genomic structure of the AML1 gene involved in acute myeloid leukemia. , 1995, Nucleic acids research.

[31]  H. Prosser,et al.  A phorbol ester response element within the human T-cell receptor beta-chain enhancer. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[32]  E. Scott,et al.  Requirement of transcription factor PU.1 in the development of multiple hematopoietic lineages. , 1994, Science.

[33]  J. Redondo,et al.  Indistinguishable nuclear factor binding to functional core sites of the T-cell receptor delta and murine leukemia virus enhancers. , 1992, Molecular and cellular biology.

[34]  E. Wieschaus,et al.  The localized requirements for a gene affecting segmentation in Drosophila: analysis of larvae mosaic for runt. , 1985, Developmental biology.

[35]  A. Look,et al.  Identification of a region which directs the monocytic activity of the colony-stimulating factor 1 (macrophage colony-stimulating factor) receptor promoter and binds PEBP2/CBF (AML1) , 1994, Molecular and cellular biology.

[36]  J. Rowley,et al.  Consistent intergenic splicing and production of multiple transcripts between AML1 at 21q22 and unrelated genes at 3q26 in (3;21)(q26;q22) translocations. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[37]  G. Keller,et al.  Rescue of erythroid development in gene targeted GATA–1− mouse embryonic stem cells , 1992, Nature Genetics.

[38]  N. Lenny,et al.  The t(8;21) fusion protein interferes with AML-1B-dependent transcriptional activation , 1995, Molecular and cellular biology.

[39]  G. Keller,et al.  Novel insights into erythroid development revealed through in vitro differentiation of GATA-1 embryonic stem cells. , 1994, Genes & development.

[40]  D. Le Paslier,et al.  The t(12;21) of acute lymphoblastic leukemia results in a tel-AML1 gene fusion. , 1995, Blood.

[41]  J. Downing,et al.  An AML1/ETO fusion transcript is consistently detected by RNA-based polymerase chain reaction in acute myelogenous leukemia containing the (8;21)(q22;q22) translocation. , 1993, Blood.

[42]  F. Collins,et al.  The leukemic core binding factor beta-smooth muscle myosin heavy chain (CBF beta-SMMHC) chimeric protein requires both CBF beta and myosin heavy chain domains for transformation of NIH 3T3 cells. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[43]  G. Dranoff,et al.  Involvement of granulocyte-macrophage colony-stimulating factor in pulmonary homeostasis. , 1994, Science.

[44]  B. Wieringa,et al.  Targeting of the creatine kinase M gene in embryonic stem cells using isogenic and nonisogenic vectors. , 1992, Nucleic acids research.

[45]  M. Ohki,et al.  t(8;21) breakpoints on chromosome 21 in acute myeloid leukemia are clustered within a limited region of a single gene, AML1. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[46]  Stuart H. Orkin,et al.  An early haematopoietic defect in mice lacking the transcription factor GATA-2 , 1994, Nature.

[47]  P. Watkins,et al.  Transcriptionally active chimeric gene derived from the fusion of the AML1 gene and a novel gene on chromosome 8 in t(8;21) leukemic cells. , 1992, Cancer genetics and cytogenetics.

[48]  S. Hiebert,et al.  Indirect and direct disruption of transcriptional regulation in cancer: E2F and AML-1. , 1995, Critical reviews in eukaryotic gene expression.

[49]  J. Rowley,et al.  The 3;21 translocation in myelodysplasia results in a fusion transcript between the AML1 gene and the gene for EAP, a highly conserved protein associated with the Epstein-Barr virus small RNA EBER 1. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[50]  S. Misawa,et al.  Actions of human interleukin-4/B-cell stimulatory factor-1 on proliferation and differentiation of enriched hematopoietic progenitor cells in culture. , 1990, Blood.

[51]  S. Swerdlow,et al.  A functional c-myb gene is required for normal murine fetal hepatic hematopoiesis , 1991, Cell.

[52]  Yoshiaki Ito,et al.  Molecular Cloning and Characterization of PEBP2β, the Heterodimeric Partner of a Novel Drosophila runt-Related DNA Binding Protein PEBP2α , 1993 .

[53]  M. Wiles,et al.  Hematopoietic commitment during embryonic stem cell differentiation in culture. , 1993, Molecular and cellular biology.

[54]  H. Drabkin,et al.  Identification of breakpoints in t(8;21) acute myelogenous leukemia and isolation of a fusion transcript, AML1/ETO, with similarity to Drosophila segmentation gene, runt. , 1992, Blood.

[55]  A. Smith,et al.  Buffalo rat liver cells produce a diffusible activity which inhibits the differentiation of murine embryonal carcinoma and embryonic stem cells. , 1987, Developmental biology.

[56]  J. Downing,et al.  Identification of AML-1 and the (8;21) translocation protein (AML-1/ETO) as sequence-specific DNA-binding proteins: the runt homology domain is required for DNA binding and protein-protein interactions , 1993, Molecular and cellular biology.

[57]  N. Manley,et al.  Two factors that bind to highly conserved sequences in mammalian type C retroviral enhancers , 1993, Journal of virology.

[58]  A. Dunn,et al.  Granulocyte/macrophage colony-stimulating factor-deficient mice show no major perturbation of hematopoiesis but develop a characteristic pulmonary pathology. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[59]  M. Ohki,et al.  The t(8;21) translocation in acute myeloid leukemia results in production of an AML1‐MTG8 fusion transcript. , 1993, The EMBO journal.

[60]  N. Speck,et al.  Cloning and characterization of subunits of the T-cell receptor and murine leukemia virus enhancer core-binding factor , 1993, Molecular and cellular biology.

[61]  M. Krangel,et al.  c-Myb and core-binding factor/PEBP2 display functional synergy but bind independently to adjacent sites in the T-cell receptor delta enhancer , 1995, Molecular and cellular biology.

[62]  Y. Yamaguchi-Iwai,et al.  Isolation of PEBP2 alpha B cDNA representing the mouse homolog of human acute myeloid leukemia gene, AML1. , 1993, Oncogene.

[63]  E. Macintyre,et al.  High frequency of t(12;21) in childhood B-lineage acute lymphoblastic leukemia. , 1995, Blood.

[64]  G. Keller,et al.  Multiple hematopoietic lineages develop from embryonic stem (ES) cells in culture. , 1991, Development.

[65]  C. Pui.  Medical progress : childhood leukemias , 1995 .

[66]  J. Gergen,et al.  The Drosophila segmentation gene runt encodes a novel nuclear regulatory protein that is also expressed in the developing nervous system. , 1990, Genes & development.

[67]  S. Nishikawa,et al.  The murine mutation osteopetrosis is in the coding region of the macrophage colony stimulating factor gene , 1990, Nature.

[68]  N. Speck,et al.  Characterization of a protein that binds multiple sequences in mammalian type C retrovirus enhancers , 1993, Journal of virology.

[69]  K. Arai,et al.  Positive and negative regulation of granulocyte-macrophage colony-stimulating factor promoter activity by AML1-related transcription factor, PEBP2. , 1995, Blood.

[70]  S. Orkin,et al.  Erythroid differentiation in chimaeric mice blocked by a targeted mutation in the gene for transcription factor GATA-1 , 1991, Nature.

[71]  C. Eaves,et al.  Properties of the earliest clonogenic hemopoietic precursors to appear in the developing murine yolk sac. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[72]  J. Gergen,et al.  The Drosophila segmentation gene runt acts as a position-specific numerator element necessary for the uniform expression of the sex-determining gene Sex-lethal. , 1991, Genes & development.

[73]  S. Shurtleff,et al.  Frequent deletion of p16INK4a/MTS1 and p15INK4b/MTS2 in pediatric acute lymphoblastic leukemia. , 1995, Blood.

[74]  N. Speck,et al.  The role of viral enhancer "core" motif-related sequences in regulating T cell receptor-gamma and -delta gene expression. , 1993, Journal of immunology.

[75]  S. Orkin,et al.  Absence of blood formation in mice lacking the T-cell leukaemia oncoprotein tal-1/SCL , 1995, Nature.

[76]  P. Sperryn,et al.  Blood. , 1989, British journal of sports medicine.

[77]  T. Rabbitts,et al.  Chromosomal translocations in human cancer , 1994, Nature.

[78]  M. Evans,et al.  The Oncogenic Cysteine-rich LIM domain protein Rbtn2 is essential for erythroid development , 1994, Cell.

[79]  F. Calabi,et al.  Leukaemia/Drosophila homology , 1992, Nature.

[80]  A. Dunn,et al.  Mice lacking both macrophage- and granulocyte-macrophage colony-stimulating factor have macrophages and coexistent osteopetrosis and severe lung disease. , 1994, Blood.

[81]  E. Golemis,et al.  Mutation of the core or adjacent LVb elements of the Moloney murine leukemia virus enhancer alters disease specificity. , 1990, Genes & development.

[82]  James Douglas Engel,et al.  Targeted disruption of the GATA3 gene causes severe abnormalities in the nervous system and in fetal liver haematopoiesis , 1995, Nature Genetics.

[83]  Y. Yamaguchi-Iwai,et al.  Expression of the Runt domain-encoding PEBP2 alpha genes in T cells during thymic development , 1995, Molecular and cellular biology.