CD34 ؉ hematopoietic stem cells are used clinically to support cytotoxic therapy, and recent studies raised hope that they could even serve as a cellular source for nonhematopoietic tissue engineering. Here, we examined in 18 volunteers the gene expressions of 1185 genes in highly enriched bone marrow CD34 ؉ (BM-CD34 ؉ ) or granulocyte-colony-stimulating factormobilized peripheral blood CD34 ؉ (PB-CD34 ؉ ) cells by means of cDNA array technology to identify molecular causes underlying the functional differences between circulating and sedentary hematopoietic stem and progenitor cells. In total, 65 genes were significantly differentially expressed. Greater cell cycle and DNA synthesis activity of BM-CD34 ؉ than PB-CD34 ؉ cells were reflected by the 2-to 5-fold higher expression of 9 genes involved in cell cycle progression, 11 genes regulating DNA synthesis, and cell cycleinitiating transcription factor E2F-1. Conversely, 9 other transcription factors, including the differentiation blocking GATA2 and N-myc, were expressed 2 to 3 times higher in PB-CD34 ؉ cells than in BM-CD34 ؉ cells. Expression of 5 apoptosis driving genes was also 2 to 3 times greater in PB-CD34 ؉ cells, reflecting a higher apoptotic activity. In summary, our study provides a gene expression profile of primary human CD34 ؉ hematopoietic cells of the blood and marrow. Our data molecularly confirm and explain the finding that CD34 ؉ cells residing in the bone marrow cycle more rapidly, whereas circulating CD34 ؉ cells consist of a higher number of quiescent stem and progenitor cells. Moreover, our data provide novel molecular insight into stem cell physiology.

Recently, overlapping molecular phenotypes of hematopoietic and neuropoietic cells were described in mice. Here, we examined primary human CD34 ؉ hematopoietic stem and progenitor cells applying specialized cDNA arrays, real-time reverse-transcriptase-polymerase chain reaction (RT-PCR), and fluorescent-activated cell sorter (FACS) analysis focusing on genes involved in neurobiologic functions. We found expression of vesicle fusion and motility genes, ligand-and voltage-gated ion channels, receptor kinases and phosphatases, and, most interestingly, mRNA as well as protein expression of G protein-coupled receptors of neuromediators (corticotropin-releasing hormone 1 [CRH 1] and CRH 2 receptors, orexin/ hypocretin 1 and 2 receptors, GABA B receptor, adenosine A 2 B receptor, opioid 1 and 1 receptors, and 5-HT 1F receptor). As shown by 2-color immunofluorescence, the protein expression of these receptors was higher in the more immature CD38 dim than in the CD38 bright subset within the CD34 ؉ population, and completely absent in fully differentiated blood cells, suggesting that those receptors play a role in developmentally early CD34 ؉ stem and progenitor cells. IntroductionHuman CD34 ϩ hematopoietic stem and progenitor cells ensure lifelong production of mature blood cells according to the varying needs of the individual. Hematopoiesis is a precisely regulated process based upon a balance of self-renewal and commitment to differentiation along the different hematopoietic lineages. The restorative capacity of human CD34 ϩ cells is clinically used in the autologous and allogeneic transplantation setting to reconstitute hematopoiesis following cytotoxic therapy for the treatment of patients with malignant or autoimmune diseases. [1][2][3] Beyond that, data of recent studies suggest that hematopoietic progenitors might also be able to transdifferentiate into nonhematopoietic cells, which could open novel therapeutic avenues in the treatment of diseases such as myocardial or cerebral infarction as well as other degenerative disorders. [4][5][6][7] However, novel data have challenged the transdifferentiation model by suggesting cell fusion rather than plasticity of stem cells. [8][9][10] A better molecular understanding of the signal perception pathways of hematopoietic stem and progenitor cells seems to be required to understand the conditions under which transdifferentiation of hematopoietic cells may occur. 11 Studies in animal models showed the presence of sensory and autonomic nerves in the bone marrow as a morphologic correlate of a possible neural regulation of hematopoiesis. [12][13][14] However, the idea that neuromediators might directly influence hematopoietic progenitors is controversially discussed. [15][16][17][18][19] Recently, several investigators described partly overlapping genetic programs of hematopoietic and neuropoietic cells in mice. 20,21 Those findings prompted us to examine human hematopoietic cells by means of specialized cDNA arrays, quantitative real-time reversetranscription-polymerase ch...

Chronic myelogenous leukemia (CML) is a malignant disorder of the hematopoietic stem cell characterized by the BCR-ABL oncogene. We examined gene expression profiles of highly enriched CD34 þ hematopoietic stem and progenitor cells from patients with CML in chronic phase using cDNA arrays covering 1.185 genes. Comparing CML CD34 þ cells with normal CD34 þ cells, we found 158 genes which were significantly differentially expressed. Gene expression patterns reflected BCR-ABLinduced functional alterations such as increased cell-cycle and proteasome activity. Detoxification enzymes and DNA repair proteins were downregulated in CML CD34 þ cells, which might contribute to genetic instability. Decreased expression of junction plakoglobulin and CXC chemokine receptor 4 (CXCR-4) might facilitate the release of immature precursors from bone marrow in CML. GATA-2 was upregulated in CML CD34 þ cells, suggesting an increased self-renewal in comparison with normal CD34 þ cells. Moreover, we found upregulation of the proto-oncogene SKI and of receptors for neuromediators such as opioid l1 receptor, GABA B receptor, adenosine A1 receptor, orexin 1 and 2 receptors and corticotropine-releasing hormone receptor. Treatment of CML progenitor cells with the selective adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) resulted in a dose-dependent significant inhibition of clonogenic growth by 40% at a concentration of 10 À5 M, which could be reversed by the equimolar addition of the receptor agonist 2-chloro-N6-cyclopentyladenosine (Po0.05). The incubation of normal progenitor cells with DPCPX resulted in an inhibition of clonogenic growth to a significantly lesser extent in comparison with CML cells (Po0.05), suggesting that the adenosine A1 receptor is of functional relevance in CML hematopoietic progenitor cells.

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