Medial HOXA genes demarcate haematopoietic stem cell fate during human development

Diana R Dou; Vincenzo Calvanese; Maria I Sierra; Andrew T Nguyen; Arazin Minasian; Pamela Saarikoski; Rajkumar Sasidharan; Christina M Ramirez; Jerome A Zack; Gay M Crooks; Zoran Galic; Hanna K A Mikkola

doi:10.1038/ncb3354

Medial HOXA genes demarcate haematopoietic stem cell fate during human development

Nat Cell Biol. 2016 Jun;18(6):595-606. doi: 10.1038/ncb3354. Epub 2016 May 16.

Authors

Diana R Dou^{1

2

3}, Vincenzo Calvanese^{1

2}, Maria I Sierra^{1

2}, Andrew T Nguyen¹, Arazin Minasian^{1

2}, Pamela Saarikoski^{1

2}, Rajkumar Sasidharan^{1

2}, Christina M Ramirez⁴, Jerome A Zack^{2

5

6}, Gay M Crooks^{1

2

7}, Zoran Galic^{2

5}, Hanna K A Mikkola^{1

2

3}

Affiliations

¹ Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, Los Angeles, California 90095, USA.
² Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, California 90095, USA.
³ Molecular Biology Institute, University of California, Los Angeles, Los Angeles, California 90095, USA.
⁴ Department of Biostatistics, Fielding School of Public Health, University of California, Los Angeles, Los Angeles, California 90095, USA.
⁵ Department of Medicine, Division of Hematology-Oncology, University of California, Los Angeles, Los Angeles, California 90095, USA.
⁶ Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, California 90095, USA.
⁷ Department of Pathology and Laboratory Medicine, University of California, Los Angeles, Los Angeles, California 90095, USA.

PMID: 27183470
PMCID: PMC4981340
DOI: 10.1038/ncb3354

Abstract

Pluripotent stem cells (PSCs) may provide a potential source of haematopoietic stem/progenitor cells (HSPCs) for transplantation; however, unknown molecular barriers prevent the self-renewal of PSC-HSPCs. Using two-step differentiation, human embryonic stem cells (hESCs) differentiated in vitro into multipotent haematopoietic cells that had the CD34(+)CD38(-/lo)CD90(+)CD45(+)GPI-80(+) fetal liver (FL) HSPC immunophenotype, but exhibited poor expansion potential and engraftment ability. Transcriptome analysis of immunophenotypic hESC-HSPCs revealed that, despite their molecular resemblance to FL-HSPCs, medial HOXA genes remained suppressed. Knockdown of HOXA7 disrupted FL-HSPC function and caused transcriptome dysregulation that resembled hESC-derived progenitors. Overexpression of medial HOXA genes prolonged FL-HSPC maintenance but was insufficient to confer self-renewal to hESC-HSPCs. Stimulation of retinoic acid signalling during endothelial-to-haematopoietic transition induced the HOXA cluster and other HSC/definitive haemogenic endothelium genes, and prolonged HSPC maintenance in culture. Thus, medial HOXA gene expression induced by retinoic acid signalling marks the establishment of the definitive HSPC fate and controls HSPC identity and function.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, Non-U.S. Gov't

MeSH terms

Antigens, CD34 / metabolism
Cell Differentiation / physiology*
Cell Lineage*
Cells, Cultured
Gene Expression Profiling / methods
Genes, Homeobox / genetics*
Hematopoietic Stem Cells / cytology*
Homeodomain Proteins / genetics
Homeodomain Proteins / metabolism*
Humans
Leukocyte Common Antigens / metabolism
Multipotent Stem Cells / cytology*
Transcriptome

Substances

Antigens, CD34
Homeodomain Proteins
HoxA protein
Leukocyte Common Antigens
PTPRC protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding