title: Antagonizing Retinoic Acid Receptors Increases Myeloid Cell Production by Cultured Human Hematopoietic Stem Cells.
abstract: Activities of the retinoic acid receptor (RAR)α and RARγ are important to hematopoiesis. Here, we have investigated the effects of receptor selective agonists and antagonists on the primitive human hematopoietic cell lines KG1 and NB-4 and purified normal human hematopoietic stem cells (HSCs). Agonizing RARα (by AGN195183) was effective in driving neutrophil differentiation of NB-4 cells and this agonist synergized with a low amount (10 nM) of 1α,25-dihydroxyvitamin D3to drive monocyte differentiation of NB-4 and KG1 cells. Treatment of cultures of human HSCs (supplemented with stem cell factor ± interleukin 3) with an antagonist of all RARs (AGN194310) or of RARα (AGN196996) prolonged the lifespan of cultures, up to 55 days, and increased the production of neutrophils and monocytes. Slowing down of cell differentiation was not observed, and instead, hematopoietic stem and progenitor cells had expanded in number. Antagonism of RARγ (by AGN205728) did not affect cultures of HSCs. Studies of CV-1 and LNCaP cells transfected with RAR expression vectors and a reporter vector revealed that RARγ and RARβ are activated by sub-nM all-trans retinoic acid (EC50-0.3 nM): ~50-fold more is required for activation of RARα (EC50-16 nM). These findings further support the notion that the balance of expression and activity of RARα and RARγ are important to hematopoietic stem and progenitor cell expansion and differentiation.
authors.affiliation: Institute of Clinical Sciences, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK. g.brown@bham.ac.uk.
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Brown
authors.affiliation: Laboratory of Protein Biochemistry, Faculty of Biotechnology, University of Wroclaw, Wroclaw, Poland.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Marchwicka
authors.affiliation: Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK.
authors.forename: Alan
authors.initials: A
authors.lastname: Cunningham
authors.affiliation: Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK.
authors.forename: Kai-Michael
authors.initials: KM
authors.lastname: Toellner
authors.affiliation: Laboratory of Protein Biochemistry, Faculty of Biotechnology, University of Wroclaw, Wroclaw, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Marcinkowska
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D018922:HL-60 Cells; D000882:Haplorhini; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D022423:Myeloid Cells / Q000276:immunology*; D009504:Neutrophils / Q000166:cytology; D011485:Protein Binding; D018168:Receptors, Retinoic Acid / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000072482:Retinoic Acid Receptor alpha / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012176:Retinoids / Q000494:pharmacology; D014212:Tretinoin / Q000737:chemistry; D000097768:Retinoic Acid Receptor gamma
chemical_list: D018952:Antigens, CD34; D018168:Receptors, Retinoic Acid; D000072482:Retinoic Acid Receptor alpha; D012176:Retinoids; D014212:Tretinoin; D000097768:Retinoic Acid Receptor gamma; C492393:AGN 195183
keywords: Agonist; All-trans retinoic acid; Antagonist; Hematopoiesis; Monocytes; Neutrophils; Retinoic acid receptor