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...

Summary:The combination of Cyclosporin A (CSA) and Methotrexate (MTX) is considered to be the standard regimen for the prevention of graft-versus-host disease (GVHD) after stem cell transplantation (SCT) from HLA-identical siblings. Mycophenolate Mofetil (MMF) has been widely used for GVHD prophylaxis after nonmyeloablative SCT, but experience following myeloablative therapy is still limited. We retrospectively compared CSA/MTX and CSA/MMF in 93 patients (median age 35 years, range 17-59 years, male subjects 48, female subjects 45) with acute myeloid leukemia (n ¼ 33), myelodysplastic syndrome (MDS) (n ¼ 3), acute lymphoblastic leukemia (ALL) (n ¼ 20) or chronic myeloid leukemia (n ¼ 37) who received CSA/MMF (n ¼ 26) or CSA/MTX (n ¼ 67) as GVHD prophylaxis following high-dose therapy and allogeneic SCT from HLA-identical siblings. No statistically significant differences were found in overall survival, relapse rate, treatment-related mortality and acute or chronic GVHD. Time to myeloid recovery was significantly shorter in patients who received CSA/MMF. We conclude that the combination of CSA/MMF appears equivalent to CSA/MTX for GVHD prophylaxis in patients receiving conventional-intensity SCT from HLA-identical siblings.

Myelodysplastic syndromes (MDS) are among the most frequent hematologic malignancies. Patients have a short survival and often progress to acute myeloid leukemia. The diagnosis of MDS can be difficult; there is a paucity of molecular markers, and the pathophysiology is largely unknown. Therefore, we conducted a multicenter study investigating whether serum proteome profiling may serve as a noninvasive platform to discover novel molecular markers for MDS. We generated serum proteome profiles from 218 individuals by MS and identified a profile that distinguishes MDS from non-MDS cytopenias in a learning sample set. This profile was validated by testing its ability to predict MDS in a first independent validation set and a second, prospectively collected, independent validation set run 5 months apart. Accuracy was 80.5% in the first and 79.0% in the second validation set. Peptide mass fingerprinting and quadrupole TOF MS identified two differential proteins: CXC chemokine ligands 4 (CXCL4) and 7 (CXCL7), both of which had significantly decreased serum levels in MDS, as confirmed with independent antibody assays. Western blot analyses of platelet lysates for these two platelet-derived molecules revealed a lack of CXCL4 and CXCL7 in MDS. Subtype analyses revealed that these two proteins have decreased serum levels in advanced MDS, suggesting the possibility of a concerted disturbance of transcription or translation of these chemokines in advanced MDS.biomarker ͉ chemokine ͉ proteomics ͉ hematologic malignancy