Antagonizing Retinoic Acid Receptors Increases Myeloid Cell Production by Cultured Human Hematopoietic Stem Cells

Activities of the retinoic acid receptor (RAR)alpha and RAR gamma 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 RAR alpha (by AGN195183) was effective in driving neutrophil differentiation of NB-4 cells and this agonist synergized with a low amount (10 nM) of 1 alpha, 25-dihydroxyvitamin D-3 to 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 RAR alpha (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 RAR gamma (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 RAR gamma and RAR beta are activated by sub-nM all-trans retinoic acid (EC50-0.3 nM): similar to 50-fold more is required for activation of RARa (EC50-16 nM). These findings further support the notion that the balance of expression and activity of RAR alpha and RAR gamma are important to hematopoietic stem and progenitor cell expansion and differentiation.