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Erythropoiesis has been extensively studied using in vitro and in vivo animal models. Despite this, there is still limited data about the gene expression profiles (GEP) of primary (ex vivo) normal human bone marrow (BM) erythroid maturation. We investigated the GEP of nucleated red blood cell (NRBC) precursors during normal human BM erythropoiesis. Three maturation-associated populations of NRBC were identified and purified from (fresh) normal human BM by flow cytometry and the GEP of each purified cell population directly analyzed using DNA-oligonucleotide microarrays. Overall, 6569 genes (19% of the genes investigated) were expressed in ≥1 stage of BM erythropoiesis at stable (e.g., genes involved in DNA process, cell signaling, protein organization and hemoglobin production) or variable amounts (e.g., genes related to cell differentiation, apoptosis, metabolism), the latter showing a tendency to either decrease from stage 1 to 3 (genes associated with regulation of erythroid differentiation and survival, e.g.,  SPI1 ,  STAT5A ) or increase from stage 2 to stage 3 (genes associated with autophagy, erythroid functions such as heme production, e.g.,  ALAS1 , ALAS2), iron metabolism (e.g.,  ISCA1, SLC11A2 ), protection from oxidative stress (e.g.,  UCP2 ,  PARK7 ), and NRBC enucleation (e.g.,  ID2 ,  RB1 ). Interestingly, genes involved in apoptosis (e.g.,  CASP8, P2RX1 ) and immune response (e.g.,  FOXO3, TRAF6 ) were also upregulated in the last stage (stage 3) of maturation of NRBC precursors. Our results confirm and extend on previous observations and providing a frame of reference for better understanding the critical steps of human erythroid maturation and its potential alteration in patients with different clonal and non-clonal erythropoietic disorders.

Maturation-associated gene expression profiles during normal human bone marrow erythropoiesis