Enforced Expression of HOXB4 in Human Embryonic Stem Cells Enhances the Production of Hematopoietic Progenitors but Has No Effect on the Maturation of Red Blood Cells
Identifieur interne : 002B77 ( Pmc/Curation ); précédent : 002B76; suivant : 002B78Enforced Expression of HOXB4 in Human Embryonic Stem Cells Enhances the Production of Hematopoietic Progenitors but Has No Effect on the Maturation of Red Blood Cells
Auteurs : Melany Jackson ; Rui Ma ; A. Helen Taylor ; Richard A. Axton ; Jennifer Easterbrook ; Maria Kydonaki ; Emmanuel Olivier ; Lamin Marenah ; Edouard G. Stanley ; Andrew G. Elefanty ; Joanne C. Mountford ; Lesley M. ForresterSource :
- Stem Cells Translational Medicine [ 2157-6564 ] ; 2016.
Abstract
Translation of a robust differentiation protocol capable of producing scalable quantities of red blood cells (RBCs) from human pluripotent stem cells to the clinic was compromised because the RBCs produced were not fully mature, expressed embryonic and fetal globins, and did not enucleate efficiently. Programming with HOXB4 increased hematopoietic progenitor and immature erythroid cell production but could not resolve the inherent challenges associated with mature adult-like enucleated RBC production.
Url:
DOI: 10.5966/sctm.2015-0324
PubMed: 27352929
PubMed Central: 4954454
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Enforced Expression of HOXB4 in Human Embryonic Stem Cells Enhances the Production of Hematopoietic Progenitors but Has No Effect on the Maturation of Red Blood Cells</title>
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<front><div type="abstract" xml:lang="en"><p>Translation of a robust differentiation protocol capable of producing scalable quantities of red blood cells (RBCs) from human pluripotent stem cells to the clinic was compromised because the RBCs produced were not fully mature, expressed embryonic and fetal globins, and did not enucleate efficiently. Programming with HOXB4 increased hematopoietic progenitor and immature erythroid cell production but could not resolve the inherent challenges associated with mature adult-like enucleated RBC production.</p>
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<front><journal-meta><journal-id journal-id-type="nlm-ta">Stem Cells Transl Med</journal-id>
<journal-id journal-id-type="iso-abbrev">Stem Cells Transl Med</journal-id>
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<issn pub-type="epub">2157-6580</issn>
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<article-meta><article-id pub-id-type="pmid">27352929</article-id>
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<article-id pub-id-type="doi">10.5966/sctm.2015-0324</article-id>
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<subject>31</subject>
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<subj-group subj-group-type="heading"><subject>Pluripotent Stem Cells</subject>
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</article-categories>
<title-group><article-title>Enforced Expression of HOXB4 in Human Embryonic Stem Cells Enhances the Production of Hematopoietic Progenitors but Has No Effect on the Maturation of Red Blood Cells</article-title>
<alt-title alt-title-type="short">HOXB4 in Erythroid Differentiation of hESCs</alt-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Jackson</surname>
<given-names>Melany</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Ma</surname>
<given-names>Rui</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Taylor</surname>
<given-names>A. Helen</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Axton</surname>
<given-names>Richard A.</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Easterbrook</surname>
<given-names>Jennifer</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Kydonaki</surname>
<given-names>Maria</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Olivier</surname>
<given-names>Emmanuel</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>b</sup>
</xref>
<xref ref-type="aff" rid="aff3"><sup>c</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Marenah</surname>
<given-names>Lamin</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>b</sup>
</xref>
<xref ref-type="aff" rid="aff3"><sup>c</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Stanley</surname>
<given-names>Edouard G.</given-names>
</name>
<xref ref-type="aff" rid="aff4"><sup>d</sup>
</xref>
<xref ref-type="aff" rid="aff5"><sup>e</sup>
</xref>
<xref ref-type="aff" rid="aff6"><sup>f</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Elefanty</surname>
<given-names>Andrew G.</given-names>
</name>
<xref ref-type="aff" rid="aff4"><sup>d</sup>
</xref>
<xref ref-type="aff" rid="aff5"><sup>e</sup>
</xref>
<xref ref-type="aff" rid="aff6"><sup>f</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Mountford</surname>
<given-names>Joanne C.</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>b</sup>
</xref>
<xref ref-type="aff" rid="aff3"><sup>c</sup>
</xref>
</contrib>
<contrib contrib-type="author" corresp="yes"><name><surname>Forrester</surname>
<given-names>Lesley M.</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
</contrib>
<aff id="aff1"><label><sup>a</sup>
</label>
Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom</aff>
<aff id="aff2"><label><sup>b</sup>
</label>
Institute of Cardiovascular and Medical Sciences, British Heart Foundation Glasgow Cardiovascular Research Centre, University of Glasgow, Glasgow, United Kingdom</aff>
<aff id="aff3"><label><sup>c</sup>
</label>
Scottish National Blood Transfusion Service, Edinburgh, United Kingdom</aff>
<aff id="aff4"><label><sup>d</sup>
</label>
Murdoch Childrens Research Institute, The Royal Children’s Hospital, Parkville, Victoria, Australia</aff>
<aff id="aff5"><label><sup>e</sup>
</label>
Department of Anatomy and Developmental Biology, Faculty of Medicine, Nursing and Health Sciences, Monash University, Clayton, Victoria, Australia</aff>
<aff id="aff6"><label><sup>f</sup>
</label>
Department of Paediatrics, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Melbourne, Victoria, Australia</aff>
</contrib-group>
<author-notes><corresp id="cor1">Correspondence: Lesley M. Forrester, Ph.D., Centre for Regenerative Medicine, University of Edinburgh, 5 Little France Drive, Edinburgh EH16 4UU, United Kingdom. Telephone: 44-<phone>131-651-9553</phone>
; E-Mail: <email>L.Forrester@ed.ac.uk</email>
</corresp>
</author-notes>
<pub-date pub-type="ppub"><month>8</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="epub"><day>28</day>
<month>6</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="pmc-release"><day>1</day>
<month>2</month>
<year>2017</year>
</pub-date>
<pmc-comment> PMC Release delay is 6 months and
0 days and was based on the . </pmc-comment>
<volume>5</volume>
<issue>8</issue>
<fpage>981</fpage>
<lpage>990</lpage>
<history><date date-type="received"><day>02</day>
<month>11</month>
<year>2015</year>
</date>
<date date-type="accepted"><day>23</day>
<month>2</month>
<year>2016</year>
</date>
</history>
<permissions><copyright-statement>©AlphaMed Press</copyright-statement>
<copyright-year>2016</copyright-year>
</permissions>
<self-uri xlink:title="pdf" xlink:type="simple" xlink:href="sctm_20150324.pdf"></self-uri>
<abstract abstract-type="precis"><p>Translation of a robust differentiation protocol capable of producing scalable quantities of red blood cells (RBCs) from human pluripotent stem cells to the clinic was compromised because the RBCs produced were not fully mature, expressed embryonic and fetal globins, and did not enucleate efficiently. Programming with HOXB4 increased hematopoietic progenitor and immature erythroid cell production but could not resolve the inherent challenges associated with mature adult-like enucleated RBC production.</p>
</abstract>
<abstract><sec><title></title>
<p>We have developed a robust, Good Manufacturing Practice-compatible differentiation protocol capable of producing scalable quantities of red blood cells (RBCs) from human pluripotent stem cells (hPSCs). However, translation of this protocol to the clinic has been compromised because the RBCs produced are not fully mature; thus, they express embryonic and fetal, rather than adult globins, and they do not enucleate efficiently. Based on previous studies, we predicted that activation of exogenous HOXB4 would increase the production of hematopoietic progenitor cells (HPCs) from hPSCs and hypothesized that it might also promote the production of more mature, definitive RBCs. Using a tamoxifen-inducible HOXB4-ER<sup>T2</sup>
expression system, we first demonstrated that activation of HOXB4 does increase the production of HPCs from hPSCs as determined by colony-forming unit culture activity and the presence of CD43<sup>+</sup>
CD34<sup>+</sup>
progenitors. Activation of HOXB4 caused a modest, but significant, increase in the proportion of immature CD235a<sup>+</sup>
/CD71<sup>+</sup>
erythroid cells. However, this did not result in a significant increase in more mature CD235a<sup>+</sup>
/CD71<sup>−</sup>
cells. RBCs produced in the presence of enhanced HOXB4 activity expressed embryonic (ε) and fetal (γ) but not adult (β) globins, and the proportion of enucleated cells was comparable to that of the control cultures. We conclude that programming with the transcription factor HOXB4 increases the production of hematopoietic progenitors and immature erythroid cells but does not resolve the inherent challenges associated with the production of mature adult-like enucleated RBCs.</p>
</sec>
<sec><title>Significance</title>
<p>As worldwide blood donations decrease and transfusable transmitted infections increase, intense interest has ensued in deriving red blood cells (RBCs) in vitro from alternative sources such as pluripotent stem cells. A translatable protocol was developed to generate RBCs; however, these RBCs have an immature phenotype. It was hypothesized that the transcription factor HOXB4 could enhance their production and maturation. Although HOXB4 increased the production of erythroid progenitors, it did not promote their maturation. Despite the remaining challenges, a robust system has been established to test other candidates and add to the knowledge base in this field.</p>
</sec>
</abstract>
<kwd-group><kwd>CD34<sup>+</sup>
</kwd>
<kwd>Cell surface markers</kwd>
<kwd>Colony formation</kwd>
<kwd>Differentiation</kwd>
<kwd>Electroporation</kwd>
<kwd>Embryonic stem cells</kwd>
<kwd>Erythroid differentiation</kwd>
<kwd>Transcription factor</kwd>
</kwd-group>
<counts><page-count count="10"></page-count>
</counts>
</article-meta>
</front>
</pmc>
</record>
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