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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.

TitleEnforced 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.
Publication TypeJournal Article
Year of Publication2016
AuthorsJackson M, Ma R, A Taylor H, Axton RA, Easterbrook J, Kydonaki M, Olivier E, Marenah L, Stanley EG, Elefanty AG, Mountford JC, Forrester LM
JournalStem Cells Transl Med
Volume5
Issue8
Pagination981-90
Date Published2016 Aug
ISSN2157-6564
KeywordsBiomarkers, Cell Differentiation, Cell Lineage, Cell Proliferation, Cells, Cultured, Cellular Reprogramming, Cellular Reprogramming Techniques, Embryonic Stem Cells, Erythrocytes, Gene Expression Regulation, Developmental, Hematopoietic Stem Cells, Homeodomain Proteins, Humans, Phenotype, Time Factors, Transcription Factors, Transfection, Up-Regulation
Abstract

UNLABELLED: : 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(T2) 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(+)CD34(+) progenitors. Activation of HOXB4 caused a modest, but significant, increase in the proportion of immature CD235a(+)/CD71(+) erythroid cells. However, this did not result in a significant increase in more mature CD235a(+)/CD71(-) 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.

SIGNIFICANCE: 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.

DOI10.5966/sctm.2015-0324
Alternate JournalStem Cells Transl Med
PubMed ID27352929
PubMed Central IDPMC4954454
Grant List / / Wellcome Trust / United Kingdom
Publication institute
CRM