Erythropoiesis from Human Embryonic Stem Cells Through Erythropoietin-Independent AKT Signaling

Authors

  • William S. Kim,

    1. Department of Pathology and Laboratory Medicine, University of California Los Angeles (UCLA),, Los Angeles, California, USA
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  • Yuhua Zhu,

    1. Department of Pathology and Laboratory Medicine, University of California Los Angeles (UCLA),, Los Angeles, California, USA
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  • Qiming Deng,

    1. Department of Biological Chemistry, David Geffen School of Medicine, University of California Los Angeles (UCLA), Los Angeles, California, USA
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  • Chee Jia Chin,

    1. Department of Pathology and Laboratory Medicine, University of California Los Angeles (UCLA),, Los Angeles, California, USA
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  • Chong Bin He,

    1. Department of Pathology and Laboratory Medicine, University of California Los Angeles (UCLA),, Los Angeles, California, USA
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  • Amanda J. Grieco,

    1. Einstein Center for Human Embryonic Stem Cell Research, Department of Medicine, Hematology and Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York, USA
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  • Gautam G. Dravid,

    1. Department of Pathology and Laboratory Medicine, University of California Los Angeles (UCLA),, Los Angeles, California, USA
    2. Stem Cell Technologies, Vancouver, British Colombia, Canada
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  • Chintan Parekh,

    1. Department of Pediatrics, Children's Hospital Los Angeles, University of Southern California Keck School of Medicine, Los Angeles, CA, USA
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  • Roger P. Hollis,

    1. Department of Microbiology, Immunology and Molecular Genetics, University of California Los Angeles (UCLA), Los Angeles, California, USA
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  • Timothy F. Lane,

    1. Department of Biological Chemistry, David Geffen School of Medicine, University of California Los Angeles (UCLA), Los Angeles, California, USA
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  • Eric E. Bouhassira,

    1. Einstein Center for Human Embryonic Stem Cell Research, Department of Medicine, Hematology and Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York, USA
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  • Donald B. Kohn,

    1. Department of Microbiology, Immunology and Molecular Genetics, University of California Los Angeles (UCLA), Los Angeles, California, USA
    2. Broad Stem Cell Research Center, University of California Los Angeles (UCLA), Los Angeles, California, USA
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  • Gay M. Crooks

    Corresponding author
    1. Department of Pathology and Laboratory Medicine, University of California Los Angeles (UCLA),, Los Angeles, California, USA
    2. Broad Stem Cell Research Center, University of California Los Angeles (UCLA), Los Angeles, California, USA
    • Correspondence: Gay M. Crooks, M.B., B.S., Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California Los Angeles, TLSB 3014, 610 Charles E Young Drive, East Los Angeles, CA 90095-1732, USA. Telephone: 310-206-0205; Fax: 310-206-0356; e-mail: gcrooks@mednet.ucla.edu

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Abstract

Unlimited self renewal capacity and differentiation potential make human pluripotent stem cells (PSC) a promising source for the ex vivo manufacture of red blood cells (RBCs) for safe transfusion. Current methods to induce erythropoiesis from PSC suffer from low yields of RBCs, most of which are immature and contain embryonic and fetal rather than adult hemoglobins. We have previously shown that homodimerization of the intracellular component of MPL (ic-MPL) induces erythropoiesis from human cord blood progenitors. The goal of this study was to investigate the potential of ic-MPL dimerization to induce erythropoiesis from human embryonic stem cells (hESCs) and to identify the signaling pathways activated by this strategy. We present here the evidence that ic-MPL dimerization induces erythropoietin (EPO)-independent erythroid differentiation from hESC by inducing the generation of erythroid progenitors and by promoting more efficient erythroid maturation with increased RBC enucleation as well as increased gamma:epsilon globin ratio and production of beta-globin protein. ic-MPL dimerization is significantly more potent than EPO in inducing erythropoiesis, and its effect is additive to EPO. Signaling studies show that dimerization of ic-MPL, unlike stimulation of the wild type MPL receptor, activates AKT in the absence of JAK2/STAT5 signaling. AKT activation upregulates GATA-1 and FOXO3 transcriptional pathways with resulting inhibition of apoptosis, modulation of cell cycle, and enhanced maturation of erythroid cells. These findings open up potential new targets for the generation of therapeutically relevant RBC products from hPSC. Stem Cells 2014;32:1503–1514

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