Differential expression of cell adhesion molecules by human hematopoietic progenitor cells from bone marrow and mobilized adult peripheral blood

Stem Cells ◽  
1995 ◽  
Vol 13 (3) ◽  
pp. 311-316 ◽  
Author(s):  
Marc L. Turner ◽  
Robert S. Anthony ◽  
Alistair C. Parker ◽  
Kathleen Mcilwaine
Keyword(s):  
Bone Marrow ◽  
Cell Adhesion ◽  
Progenitor Cells ◽  
Differential Expression ◽  
Adhesion Molecules ◽  
Peripheral Blood ◽  
Cell Adhesion Molecules ◽  
Hematopoietic Progenitor Cells ◽  
Hematopoietic Progenitor ◽  
Adult Peripheral Blood

scholarly journals Oncogenic Deregulation of Cell Adhesion Molecules in Leukemia

Cancers ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 311 ◽  
Author(s):  
Roland Windisch ◽  
Nina Pirschtat ◽  
Christian Kellner ◽  
Linping Chen-Wichmann ◽  
Jörn Lausen ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Adhesion ◽  
Progenitor Cells ◽  
Adhesion Molecules ◽  
Cell Adhesion Molecules ◽  
Gene Products ◽  
Hematopoietic Stem ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Cell Matrix Interactions

Numerous cell–cell and cell–matrix interactions within the bone marrow microenvironment enable the controlled lifelong self-renewal and progeny of hematopoietic stem and progenitor cells (HSPCs). On the cellular level, this highly mutual interaction is granted by cell adhesion molecules (CAMs) integrating differentiation, proliferation, and pro-survival signals from the surrounding microenvironment to the inner cell. However, cell–cell and cell–matrix interactions are also critically involved during malignant transformation of hematopoietic stem/progenitor cells. It has become increasingly apparent that leukemia-associated gene products, such as activated tyrosine kinases and fusion proteins resulting from chromosomal translocations, directly regulate the activation status of adhesion molecules, thereby directing the leukemic phenotype. These observations imply that interference with adhesion molecule function represents a promising treatment strategy to target pre-leukemic and leukemic lesions within the bone marrow niche. Focusing on myeloid leukemia, we provide a current overview of the mechanisms by which leukemogenic gene products hijack control of cellular adhesion to subsequently disturb normal hematopoiesis and promote leukemia development.

The RNA Editing Enzyme ADAR1 Is Required for Survival of Differentiating Hematopoietic Progenitor Cells in Adult Mice.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1395-1395
Author(s):  
Feng Xu ◽  
Qingde Wang ◽  
Hongmei Shen ◽  
Hui Yu ◽  
Yanxin Li ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Rna Editing ◽  
Cell Population ◽  
Peripheral Blood ◽  
Hematopoietic Progenitor Cells ◽  
Hematopoietic Progenitor ◽  
Hematopoietic Stem ◽  
Low Level ◽  
Adult Mice

Abstract Adenosine Deaminases Acting on RNA (ADAR) are RNA-editing enzymes converting adenosine residues into inosine (A-to-I) in many double-stranded RNA substrates including coding and non-coding sequences as well as microRNAs. Disruption of the ADAR1 gene in mice results in fetal liver, but not yolk sac, defective erythropoiesis and death at E11.5 (Wang Q et al, Science 2000). Subsequently, a conditional knockout mouse model confirmed these findings and showed massively increased cell death in the affected organs (Wang Q et al, JBC 2004). However, the actual impact of ADAR1 absence on definitive or adult hematopoiesis has not been examined. To define the role of ADAR1 in adult hematopoiesis, we first examined the expression of ADAR1 in different hematopoietic stem/progenitor cell subsets isolated from bone marrow by real-time RT-PCR. ARAR1 was present in hematopoietic stem cells (HSCs) at relatively low level and increased in hematopoietic progenitor cells (HPCs). A series of functional hematopoietic assays were then undertaken. A conditional deletion of ADAR1 was achieved by transducing Lin− or Lin−cKit+ bone marrow cells from ADAR1-lox/lox mice with a MSCV retroviral vector co-expressing Cre and GFP. PCR analysis confirmed the complete deletion of ADAR1 in the transduced cells within 72 hours after the transduction. This system allowed us to evaluate the acute effect of ADAR1 deletion in a specific hematopoietic cell population. Following 4 days of in vitro culture after transduction, the absolute number of Lin− Sca1+ cells in the Cre transduced group was similar to the input number; however the differentiating Lin+ cells significantly decreased whereas both the Lin−Sca1+ and Lin+ cells in the vector (MSCV carrying GFP alone) transduced group increased during culture. Moreover, the colony forming cell (CFC) assay showed much fewer and smaller colonies that contained dead cells from the gene deleted group as compared to those from the control group (p<0.001). The TUNEL assay showed a dramatic increase of apoptosis in the Lin+ population but not in the Lin− cells. Given the mixed genetic background of the ADAR1-lox/lox mice, repopulation of the transduced hematopoietic cells in vivo was examined in immunodeficient mice. Sublethally irradiated (3.5 Gy) NOD/SCID-γcnull recipient were transplanted with either 1.5 × 105 Cre or vector transduced Lin− ADAR1-lox/lox cells. Multi-lineage engraftment in peripheral blood was monitored monthly. While the vector transduced cells were able to constitute more than 90% in multiple lineages of the peripheral blood at 1 to 3 months, Cre-transduced cells were virtually undetectable at all the time points (n=9 to 13, p<0.001). A similar result was found in the hematopoietic organs, including the bone marrow, spleen and thymus. Interestingly, however, the Lin−Sca1+cKit+ cell population was preserved in the Cre transduced group despite the very low level of total donor-derived cells in the bone marrow (n=6 to 7, p<0.01). Consistently, the single cell culture experiment demonstrated that there was no significant difference between ADAR−/− and wild-type HSCs in terms of survival and division during the first 3 days of culture. Taken together, our current study demonstrates nearly absolute requirement of ADAR1 for hematopoietic repopulation in adult mice and it is also suggested that ADAR1 has a preferential effect on the survival of differentiating progenitor cells over more primitive cells.

Angiogenic Factors eNOS and VEGF Have Increased Expression in Granulocytes of JAK2V617F Homozygous Forms of Polycythemia Vera,

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3847-3847
Author(s):  
Vladan P Cokic ◽  
Dragana Markovic ◽  
Olivera Mitrovic ◽  
Sanja Vignjevic ◽  
Dragoslava Djikic ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bone Marrow ◽  
Transcription Factor ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Protein Level ◽  
Angiogenic Factors ◽  
Hematopoietic Progenitor Cells ◽  
Hematopoietic Progenitor ◽  
Vegf Protein

Abstract Abstract 3847 The microvessel density of bone marrow is increased in myeloproliferative neoplasms (MPN) parallel with vascular endothelial growth factor (VEGF). VEGF-mediated angiogenesis requires nitric oxide (NO) production from activated endothelial NO synthase (eNOS). NO as well as hypoxia stimulate the VEGF gene expression and angiogenesis by enhancing hypoxia inducible factor (HIF)-1 activity. We studied 126 newly diagnosed patients with BCR-ABL− MPN: 64 polycythemia vera (PV), 36 essential thrombocythemia (ET), 26 primary myelofibrosis (PMF) and 12 healthy individuals. We performed a combined analysis of hematopoietic CD34+ progenitor cells and granulocytes in peripheral blood of these individuals. The eNOS protein level is more than three-fold elevated in granulocytes of JAK2V617F homozygous PV patients. The essential inducer of angiogenesis VEGF-A has also about three-fold elevation at the protein level in granulocytes of PV patients, with major increases in JAK2V617F homozygous forms. Immunohistochemical analysis reveal that the percentage of VEGF-A-positive cells is increased in bone marrow of PV (5.58±0.7%) compared to normal controls (2.78±0.7%) and VEGF-A mRNA levels are increased in hematopoietic progenitor cells of PV origin. Transcription factor HIF-1α gene expression is decreased in hematopoietic progenitor cells and increased in granulocytes of PV patients. Negative regulator of HIF-1α activity, a transcription factor HIF-3α, has decreased expression in hematopoietic progenitor cells and not changed in granulocytes. In contrast to PV patients, PMF and ET disorders with a minor JAK2 mutation burden demonstrate reduced eNOS and VEGF protein levels and decreased HIF-1a gene expression in peripheral blood granulocytes, although the increase in percentage of VEGF-A-positive cells in bone marrow observed in PV patients is also evident. The present results expand the significance of JAK2V617F mutation in induction of angiogenic factors eNOS and VEGF in granulocytes of PV patients with enhanced HIF-1α presence. Moreover, the stromal and hematopoietic cells also show increased VEGF protein expression in bone marrow of PV patients. Therefore, we find that variations in angiogenic factors expression among MPN patients appear to be related to JAK2V617F mutation allele burden. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Arhgap21 Expression in Bone Marrow Niche Is Crucial for Hematopoietic Progenitor Homing and Short Term Reconstitution after Transplantation

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 1591-1591
Author(s):  
Juliana M. Xavier ◽  
Lauremilia Ricon ◽  
Karla Priscila Vieira ◽  
Longhini Ana Leda ◽  
Carolina Bigarella ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Hematopoietic Progenitor Cells ◽  
Bone Marrow Niche ◽  
Wild Type ◽  
Hematopoietic Progenitor ◽  
Short Term ◽  
Hematopoietic Stem

Abstract The microenvironment of the bone marrow (BM) is essential for retention and migration of hematopoietic progenitor cells. ARHGAP21 is a negative regulator of RhoGTPAses, involved in cellular migration and adhesion, however the role of ARHGAP21 in hematopoiesis is unknown. In order to investigate whether downregulation of Arhgap21 in microenvironment modulates bone marrow homing and reconstitution, we generated Arhgap21+/-mice using Embryonic Stem cell containing a vector insertion in Arhgap21 gene obtained from GeneTrap consortium and we then performed homing and bone marrow reconstitution assays. Subletally irradiated (9.5Gy) Arhgap21+/- and wild type (WT) mice received 1 x 106 BM GFP+cells by IV injection. For homing assay, 19 hours after the transplant, Lin-GFP+ cells were analyzed by flow cytometry. In reconstitution and self-renew assays, the GFP+ cell percentage in peripheral blood were analyzed 4, 8, 12 and 16 weeks after transplantation. Hematopoietic stem cells [GFP+Lin-Sca+c-Kit+ (LSK)] were counted after 8 and 16 weeks in bone marrow after primary transplant and 16 weeks after secondary transplant. The percentage of Lin-GFP+ hematopoietic progenitor cells that homed to Arhgap21+/-recipient (mean± SD) (2.07 ± 0.85) bone marrow was lower than those that homed to the WT recipient (4.76 ± 2.60); p=0.03. In addition, we observed a reduction (WT: 4.22 ±1.39; Arhgap21+/-: 2.17 ± 0.69; p=0.001) of Lin- GFP+ cells in Arhgap21+/-receptor spleen together with an increase of Lin- GFP+ population in Arhgap21+/-receptor peripheral blood (WT: 8.07 ± 3.85; Arhgap21+/-: 14.07 ±5.20; p=0.01), suggesting that hematopoietic progenitor cells which inefficiently homed to Arhgap21+/-bone marrow and spleen were retained in the blood stream. In bone marrow reconstitution assay, Arhgap21+/-receptor presented reduced LSK GFP+ cells after 8 weeks (WT: 0.19 ±0.03; Arhgap21+/-0.12±0.05; p=0.02) though not after 16 weeks from primary and secondary transplantation. The reduced LSK percentage after short term reconstitution was reflected in the lower GFP+ cells in peripheral blood 12 weeks after transplantation (WT: 96.2 ±1.1; Arhgap21+/-94.3±1.6; p=0.008). No difference was observed in secondary transplantation, indicating that Arhgap21reduction in microenvironment does not affect normal hematopoietic stem cell self-renewal. The knowledge of the niche process in regulation of hematopoiesis and their components helps to better understand the disordered niche function and gives rise to the prospect of improving regeneration after injury or hematopoietic stem and progenitor cell transplantation. In previous studies, the majority of vascular niche cells were affected after sublethal irradiation, however osteoblasts and mesenchymal stem cells were maintained (Massimo Dominici et al.; Blood; 2009.). RhoGTPase RhoA, which is inactivated by ARHGAP21 (Lazarini et al.; Biochim Biophys acta; 2013), has been described to be crucial for osteoblasts and mesenchymal stem cell support of hematopoiesis (Raman et al.; Leukemia; 2013). Taken together, these results suggest that Arhgap21 expression in bone marrow niche is essential for homing and short term reconstitution support. Moreover, this is the first study to investigate the role of Arhgap21 in bone marrow niche. Figure 1 Reduced homing and short term reconstitution in Arhgap21 +/- recipients. Bone marrow cells from GFP+ mice were injected into wild-type and Arhgap21+/- sublethally irradiated mice. 19 hours after the transplant, a decreased homing was observed to both bone marrow (a) and spleen (b) together with an increase of retained peripheral blood (c) Lin-GFP+ cells. In serial bone marrow transplantation, Arhgap21+/- presented reduced bone marrow LSK GFP+ cells 8 weeks (d) and peripheral blood GFP+ cells 12 weeks (e) after primary transplantation, though not 16 weeks after primary (f) and 16 weeks after secondary (g) transplantations. The result is expressed by means ±SD of 2 independent experiments. Figure 1. Reduced homing and short term reconstitution in Arhgap21+/- recipients. Bone marrow cells from GFP+ mice were injected into wild-type and Arhgap21+/- sublethally irradiated mice. 19 hours after the transplant, a decreased homing was observed to both bone marrow (a) and spleen (b) together with an increase of retained peripheral blood (c) Lin-GFP+ cells. In serial bone marrow transplantation, Arhgap21+/- presented reduced bone marrow LSK GFP+ cells 8 weeks (d) and peripheral blood GFP+ cells 12 weeks (e) after primary transplantation, though not 16 weeks after primary (f) and 16 weeks after secondary (g) transplantations. The result is expressed by means ±SD of 2 independent experiments. Disclosures No relevant conflicts of interest to declare.

scholarly journals Cellularly defined minor histocompatibility antigens are differentially expressed on human hematopoietic progenitor cells.

1988 ◽  
Vol 168 (6) ◽  
pp. 2337-2347 ◽  
Author(s):  
P J Voogt ◽  
E Goulmy ◽  
W F Veenhof ◽  
M Hamilton ◽  
W E Fibbe ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Growth Inhibition ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Bone Marrow Cells ◽  
Differentially Expressed ◽  
Class I ◽  
Hematopoietic Progenitor Cells ◽  
Hematopoietic Progenitor ◽  
Immune Mediated

Previously, five CTL lines directed against minor histocompatibility (mH) antigens designated HA-1-5 have been established from peripheral blood of patients after allogeneic bone marrow transplantation (BMT), and have been characterized using population and family studies. All cell lines showed specific HLA class I-restricted lysis of PHA-stimulated peripheral blood target cells from donors positive for the particular mH antigens. After 4 h of incubation of the mH antigen HA-3-specific CTL line with bone marrow cells from HA-3+ donors, complete class I-restricted inhibition of colony growth of the hematopoietic progenitor cells was observed even at low E/T ratios, indicating that the HA-3 antigen is strongly expressed on hematopoietic stem cells. Therefore, this antigen may be a target structure in the immune-mediated rejection of the hematopoietic graft in case of incompatibility for this determinant between donor and recipient in allogeneic BMT. In contrast, incubation of bone marrow cells with the antigen-specific anti-HA-1, -2, -4, and -5 CTL lines did not result in growth inhibition of the hematopoietic progenitor cells tested. After a prolonged incubation time and using a very high E/T ratio, progenitor cells from HA-2+ or HA-5+ donors were killed to some extent by the anti-mH-specific CTL lines, although the growth inhibition observed was minor and variable. Our results show that mH antigens are differentially expressed on human hematopoietic progenitor cells. Therefore, only some of these antigens may be targets in immune-mediated rejection of the bone marrow graft.

Ultrastructural Features of CD34+Hematopoietic Progenitor Cells from Bone Marrow, Peripheral Blood and Umbilical Cord Blood

2001 ◽  
Vol 42 (4) ◽  
pp. 699-708 ◽  
Author(s):  
Giorgio Lambertenghi Deliliers ◽  
Lorenza Caneva ◽  
Rossella Fumiatti ◽  
Federica Servida ◽  
Paolo Rebulla ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cord Blood ◽  
Progenitor Cells ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Peripheral Blood ◽  
Hematopoietic Progenitor Cells ◽  
Hematopoietic Progenitor

Correction of Murine Adamts13 Deficiency by Hematopoietic Progenitor Cell-Mediated Gene Therapy

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2356-2356
Author(s):  
Pablo Laje ◽  
Dezhi Shang ◽  
Masayuki Endo ◽  
Wenjing Cao ◽  
Philip W. Zoltick ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Arterial Occlusion ◽  
Hematopoietic Progenitor Cells ◽  
Adamts13 Activity ◽  
Hematopoietic Progenitor ◽  
Adamts13 Deficiency ◽  
Carotid Arterial ◽  
Bone Marrow Chimerism

Abstract ADAMTS13, a metalloprotease that cleaves von Willebrand factor (vWF), is primarily synthesized in liver, endothelial cells and megakaryocytes/platelets. It circulates in plasma as an active protease with concentrations between 0.5 and 1.0 μg per milliliter of plasma. Proteolytic cleavage of endothelial cell bound and plasma vWF by ADAMTS13 is critical for maintaining normal hemostasis. Inability to cleave newly released unusually large vWF as a result of hereditary or acquired deficiency of ADAMTS13 activity leads to a potentially fatal syndrome, thrombotic thrombocytopenic purpura (TTP). Although plasma infusion or exchange is a proven effective therapy for TTP, the life-long plasma infusion for hereditary TTP is not without complications. In search for a better therapy, we performed an autologous transplantation of hematopoietic progenitor cells (HPCs) for correcting Adamts13 deficiency in a mouse model. The bone marrows were harvested from Adamts13−/− mice (C57BL6/129Sv) at 6–8 weeks of age and HPCs (CD48−/CD150+) were purified to a greater than 90% of purity in all cases using serial immune affinity depletion and enrichment techniques. The purified HPCs were transduced in culture with a self-inactivated lentiviral vector either encoding GFP or GFP plus murine full-length Adamts13 at 100 × MOI (multiplicity of infection) for 14–16 hours. The transduced HPCs (1 × 105) mixed with freshly prepared non-transduced bone marrow mononucleated cells (2 × 105) were then infused via tail vein into Adamts13−/− mice (recipients ) after lethal irradiation (450 cGy, twice, 3 h apart, at a dose rate of 2.2 cGy per minute). Peripheral blood was collected at 1, 3, and 5 months of post-transplantation for assessing the bone marrow chimerism and Adamts13 activity. The GFP positive cells were determined by flow cytometry and Adamts13 activity was determined by FRETS-vWF73 and GST-vWF73 peptides as described previously. We showed that all ten mice transplanted with the transduced HPCs encoding GFP plus murine Adamts13 exhibited persistent levels of plasma Adamts13 proteolytic activity, ranging from 10% to 60% of pooled normal murine plasma (PNP), despite low levels of bone marrow chimerism based on the percentage of GFP-positive mononucleated cells (~10–40%) in the peripheral blood. The expressed Adamts13 in plasma was able to cleave process unusually large vWF, resulting in a reduction of vWF multimers sizes compared with those in Adamts13−/− mice transplanted with HPCs transduced with the vector encoding GFP. Moreover, the levels of plasma Adamts13 appeared to be sufficient to protect mice against ferric chloride-induced carotid arterial thrombosis. The carotid arterial occlusion times for Adamts13−/− mice which did not undergo bone marrow transplantation were 5.6 ± 1.6 min, similar to those transplanted with HPCs transduced with the vector encoding GFP (4.3 ± 0.8 min) (mean ±SD) (p>0.05). However, the carotid arterial occlusion times were significantly prolonged in Adamts13−/− mice transplanted with the vector encoding GFP plus murine Adamts13 (15.5 ± 6.8 min). The differences between the experimental group and the control groups were all statistically very significant (p<0.01). The data suggest that hematopoietic progenitor cells can be genetically modified ex vivo and transplanted in an autologous model to provide adequate levels of functional Adamts13 metalloprotease. This success may provide basis for development of a novel therapeutic strategy to cure hereditary TTP in humans.

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