Oncogenic Gata1 causes stage-specific megakaryocyte differentiation delay

The megakaryocyte/erythroid Transient Myeloproliferative Disorder (TMD) in newborns with Down Syndrome (DS) occurs when N-terminal truncating mutations of the hemopoietic transcription factor GATA1, that produce GATA1short protein (GATA1s), are acquired early in development. Prior work has shown that murine GATA1s, by itself, causes a transient yolk sac myeloproliferative disorder. However, it is unclear where in the hemopoietic cellular hierarchy GATA1s exerts its effects to produce this myeloproliferative state. Here, through a detailed examination of hemopoiesis from murine GATA1s ES cells and GATA1s embryos we define defects in erythroid and megakaryocytic differentiation that occur relatively in hemopoiesis. GATA1s causes an arrest late in erythroid differentiation in vivo, and even more profoundly in ES-cell derived cultures, with a marked reduction of Ter-119 cells and reduced erythroid gene expression. In megakaryopoiesis, GATA1s causes a differentiation delay at a specific stage, with accumulation of immature, kit-expressing CD41hi megakaryocytic cells. In this specific megakaryocytic compartment, there are increased numbers of GATA1s cells in S-phase of cell cycle and reduced number of apoptotic cells compared to GATA1 cells in the same cell compartment. There is also a delay in maturation of these immature GATA1s megakaryocytic lineage cells compared to GATA1 cells at the same stage of differentiation. Finally, even when GATA1s megakaryocytic cells mature, they mature aberrantly with altered megakaryocyte-specific gene expression and activity of the mature megakaryocyte enzyme, acetylcholinesterase. These studies pinpoint the hemopoietic compartment where GATA1s megakaryocyte myeloproliferation occurs, defining where molecular studies should now be focussed to understand the oncogenic action of GATA1s. Scientific Category Haematopoiesis and Stem Cells Key Points GATA1s-induced stage-specific differentiation delay increases immature megakaryocytes in vivo and in vitro, during development. Differentiation delay is associated with increased numbers of cells in S-phase and reduced apoptosis.

[1]  P. Kingsley,et al.  Distinct Sources of Hematopoietic Progenitors Emerge before HSCs and Provide Functional Blood Cells in the Mammalian Embryo. , 2015, Cell reports.

[2]  J. Crispino,et al.  Global transcriptome and chromatin occupancy analysis reveal the short isoform of GATA1 is deficient for erythroid specification and gene expression , 2015, Haematologica.

[3]  R. Hardison,et al.  Pluripotent stem cells reveal erythroid-specific activities of the GATA1 N-terminus. , 2015, The Journal of clinical investigation.

[4]  Ross C Hardison,et al.  Divergent functions of hematopoietic transcription factors in lineage priming and differentiation during erythro-megakaryopoiesis , 2014, Genome research.

[5]  A. Curley,et al.  GATA1-mutant clones are frequent and often unsuspected in babies with Down syndrome: identification of a population at risk of leukemia. , 2013, Blood.

[6]  S. Orkin,et al.  Mapping cellular hierarchy by single-cell analysis of the cell surface repertoire. , 2013, Cell stem cell.

[7]  S. Miyano,et al.  The landscape of somatic mutations in Down syndrome–related myeloid disorders , 2013, Nature Genetics.

[8]  Jonghwan Kim,et al.  Developmental differences in IFN signaling affect GATA1s-induced megakaryocyte hyperproliferation. , 2013, The Journal of clinical investigation.

[9]  Cole Trapnell,et al.  TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions , 2013, Genome Biology.

[10]  Sarah Filippi,et al.  Perturbation of fetal liver hematopoietic stem and progenitor cell development by trisomy 21 , 2012, Proceedings of the National Academy of Sciences.

[11]  E. Lander,et al.  Exome sequencing identifies GATA1 mutations resulting in Diamond-Blackfan anemia. , 2012, The Journal of clinical investigation.

[12]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[13]  S. Orkin,et al.  Developmental stage-specific interplay of GATA1 and IGF signaling in fetal megakaryopoiesis and leukemogenesis. , 2010, Genes & development.

[14]  Stuart H. Orkin,et al.  GATA-2 Reinforces Megakaryocyte Development in the Absence of GATA-1 , 2009, Molecular and Cellular Biology.

[15]  Masayuki Yamamoto,et al.  Direct Binding of pRb/E2F-2 to GATA-1 Regulates Maturation and Terminal Cell Division during Erythropoiesis , 2009, PLoS biology.

[16]  P. Vyas,et al.  Characterization of megakaryocyte GATA1-interacting proteins: the corepressor ETO2 and GATA1 interact to regulate terminal megakaryocyte maturation. , 2008, Blood.

[17]  H. Nakauchi,et al.  Metalloproteinase regulation improves in vitro generation of efficacious platelets from mouse embryonic stem cells , 2008, The Journal of experimental medicine.

[18]  R. Waugh,et al.  The megakaryocyte lineage originates from hemangioblast precursors and is an integral component both of primitive and of definitive hematopoiesis. , 2007, Blood.

[19]  A. F. Cunha,et al.  An inherited mutation leading to production of only the short isoform of GATA-1 is associated with impaired erythropoiesis , 2006, Nature Genetics.

[20]  Aravind Subramanian,et al.  Identification of distinct molecular phenotypes in acute megakaryoblastic leukemia by gene expression profiling. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[21]  J. Strouboulis,et al.  A generic tool for biotinylation of tagged proteins in transgenic mice , 2005, Transgenic Research.

[22]  S. Orkin,et al.  Developmental stage–selective effect of somatically mutated leukemogenic transcription factor GATA1 , 2005, Nature Genetics.

[23]  P. Vyas,et al.  Natural history of GATA1 mutations in Down syndrome. , 2004, Blood.

[24]  A. Teigler‐Schlegel,et al.  Mutations in exon 2 of GATA1 are early events in megakaryocytic malignancies associated with trisomy 21. , 2003, Blood.

[25]  Patrick Rodriguez,et al.  Efficient biotinylation and single-step purification of tagged transcription factors in mammalian cells and transgenic mice , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[26]  M. L. Beau,et al.  Acquired mutations in GATA1 in the megakaryoblastic leukemia of Down syndrome , 2002, Nature Genetics.

[27]  J. Palis,et al.  Development of erythroid and myeloid progenitors in the yolk sac and embryo proper of the mouse. , 1999, Development.

[28]  S. Orkin,et al.  GATA-1 and erythropoietin cooperate to promote erythroid cell survival by regulating bcl-xL expression. , 1999, Blood.

[29]  S. Orkin,et al.  Consequences of GATA-1 Deficiency in Megakaryocytes and Platelets , 1999 .

[30]  S. Orkin,et al.  Improved reporter strain for monitoring Cre recombinase-mediated DNA excisions in mice. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[31]  S. Orkin,et al.  A lineage‐selective knockout establishes the critical role of transcription factor GATA‐1 in megakaryocyte growth and platelet development , 1997, The EMBO journal.

[32]  Y Fujiwara,et al.  Arrested development of embryonic red cell precursors in mouse embryos lacking transcription factor GATA-1. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[33]  W. Vainchenker,et al.  Constitutive expression of GATA-1 interferes with the cell-cycle regulation. , 1996, Blood.

[34]  R. Hardison,et al.  Pluripotent stem cells reveal erythroid-specific activities of the GATA 1 N-terminus , 2018 .

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