EFFECT OF ERYTHROPOIETIN ON CYTOKINE PRODUCTION BY STEM CELLS

Erythropoietin (EPO) is mainly used to stimulate erythropoiesis. Its cytoprotective effects upon other cells of the human body and animals were recently shown, in particular, anti-apoptotic effect was observed. EPO effect upon the cells is mediated by interaction with erythropoietin receptor, with a complex forming a heterodimeric bond with β-common chain (CD131). In the present work, we studied the changes in erythropoietin receptor expression, and production spectrum of biologically active molecules in bone marrow mononuclear cells (BM-MNC) of patients with coronary heart disease. The flow cytometric assays showed that short-term incubation of BM-MNC with erythropoietin caused increased expression of the erythropoietin receptors on hematopoietic stem cells and tended to reduce the number of endothelial progenitor cells carrying the erythropoietin receptors. Solid-phase enzyme immunoassay in conditioned media from BM-MNC revealed that long-term (72 hours) exposure of BM-MNC to erythropoietin promoted increased production of IL-1β, PDGF-AB, and Epo, if compared to the basal production level (p < 0.05). Short-term incubation of BM-MNC with erythropoietin (60 minutes) caused a significant increase in the IL-1β, PDGF-AB and CXCL-12 / SDF-1α production levels, as well as significant reduction in the IL-10 production levels compared to the basal levels (p < 0.05).

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Erythropoietin Signaling Inhibits the Short-Term Marrow Reconstitution.

Blood ◽

10.1182/blood.v106.11.3028.3028 ◽

2005 ◽

Vol 106 (11) ◽

pp. 3028-3028

Author(s):

Ko-Tung Chang ◽

Yves D. Pastore ◽

Roberto H. Nussenzveig ◽

Josef T. Prchal

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Peripheral Blood ◽

Relative Proportion ◽

Wild Type Mouse ◽

Erythropoietin Receptor ◽

Wild Type ◽

Short Term ◽

Gain Of Function ◽

Hematopoietic Stem

Abstract Murine hematopoietic stem cells (HSCs) transfected with gain-of-function human erythropoietin receptor (EPOR) transgene were reported to have a competitive advantage over wild type mouse HSCs in a bone marrow transplantation (BMT) model (Kirby, Blood; 95(12):3710, 2000). However, EpoRs may not be normally expressed in early progenitor/stem cells, moreover, whether the Epo/EpoR signaling plays a role on homing and repopulating of hematopoietic progenitor/stem cells is also unknown. Our lab has previously created a mouse model harboring either a knocked-in human wild type (wthEPOR) hypomorphic gene or a mutant human gain-of-function EPOR (mthEPOR) gene into the mouse EPOR locus (Divoky, PNAS; 98(3):986, 2001). The wthEPOR mice are anemic, while mthEpoR mice are polycythemic. We tested the possible advantage of mthEPOR HSCs in the competitive bone marrow (BM) transplantation assay using C57/Bl6 congenic mice. BM from wthEPOR (CD45.1/45.2 heterozygous or CD45.1) and mthEPOR (CD45.2) mice were co-transplanted (1:1 ratio) into lethally irradiated (137Cs >11Gy split) normal recipients (CD45.1 or CD45.1/45.2). Unexpectedly, in three independent groups the peripheral blood chimerism derived from wthEPOR-bearing cells significantly out-competed the mthEPOR-bearing cells two weeks after BMT (52% vs. 40%, 52% vs. 45% and 50% vs. 42%, n=11, 8 and 6, p<0.01). Furthermore, administration of exogenous Epo (10U/day/mouse) further impaired peripheral blood engraftment (including Ter119+ erythroid lineages) of mthEPOR cells (45% vs. 39%, n=8, p<0.05) though the relative proportion of erythroid cells within the engraftment was increased. Thus, an enhanced Epo/EpoR signaling interferes with the short-term repopulation of hematopoietic progenitors resulting in a decreased engraftment, possibly by interfering with homing. A possible long-term advantage of mthEPOR HSCs is being evaluated in ongoing studies; however, these short term data suggest that Epo administration to BM transplant patients may impede engraftment. Figure Figure

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In Vivo Treatment with Prostaglandin E2 (PGE2) Selectively Expands Short-Term Hematopoietic Stem Cells.

Blood ◽

10.1182/blood.v110.11.1254.1254 ◽

2007 ◽

Vol 110 (11) ◽

pp. 1254-1254

Author(s):

Benjamin J. Frisch ◽

Jonathan M. Weber ◽

Rebecca L. Porter ◽

Benjamin J. Gigliotti ◽

Julianne N. Smith ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Hematopoietic Stem Cells ◽

Prostaglandin E2 ◽

Mononuclear Cells ◽

Surprising Result ◽

Short Term ◽

Hematopoietic Stem ◽

Donor Cells

Abstract Parathyroid Hormone (PTH) expands hematopoietic stem cells (HSC) through activated osteoblasts in the bone marrow (BM). Since PTH stimulates osteoblastic production of Prostaglandin E2 (PGE2), we hypothesized that PGE2 could also regulate HSC. In vivo PGE2 treatment demonstrated a time and dose dependent increase in BM lineage− Sca-1+ c-kit+ (LSK) BM mononuclear cells (BMMC) from PGE2 vs. vehicle treated mice (0.11 vs. 0.04% BMMC, P=0.0061, n=8 mice per treatment group), an effect superior to PTH (350 vs. 100% increase in LSK). There were no significant PGE2 effects on CFU-Cs or peripheral Hct, Plts or WBC counts compared to vehicle. Therefore PGE2-dependent cell expansion was not global across differentiated subsets, but was restricted to primitive hematopoietic cells, similar to the effects of PTH treatment. Consistent with a PGE2-dependent HSC increase, cells from PGE2 vs vehicle-treated mice had superior lymphomyeloid reconstitution by competitive repopulation analysis. However, this increase was short-lived: specifically, PGE2-dependent myeloid (CD11b+) reconstitution was no longer superior at 6 weeks, while the PGE2-dependent increase in lymphoid (CD3e+ and B220+) reconstitution ceased by 16 weeks. This surprising result suggests that in vivo PGE2 treatment selectively expands short-term HSC (or ST-HSC), which have highly proliferative properties, but limited self-renewal. To further confirm this targeted PGE2 effect, LSK subset analysis based on Flt3 and Thy1.1 expression was performed. Consistent with the competitive repopulation data, PGE2 treatment significantly increased Flt3+Thy1.1int LSK ST-HSC (0.0273 vs 0.0140% n=4 in each group, p=0.0307) as well as Flt3+Thy1.1− LSK Multipotent Progenitors (0.0305 vs 0.0195% n=4 in each group, p=0.0070), while Flt3−Thy1.1int LSK Long-Term HSC or LT-HSC (0.0126 vs 0.0078% n=4 in each group, p=0.1069) were unchanged compared to vehicle treatment. ST vs LT-HSC activity can also be quantified by the in vivo clonogenic Colony Forming Unit-Spleen (CFU-S) assay, where day 8 CFU-S represent ST-HSC, while day 10–12 CFU-S represent LT-HSC. Consistent with a PGE2-dependent specific ST-HSC increase, BMMC from PGE2 treated mice gave rise to a significantly higher number of CFU-Sd8 compared to cells from vehicle treated mice (10.5 vs 4.75 CFU-S per 60,000 BMMC, n=4 in each group, p=0.0053), while CFU-Sd10 were unchanged (12.5 vs 11.5 CFU-S per 60,000 BMMC, n=6, p=0.4950). Finally, since ST-HSC confer radioprotection, PGE2-dependent ST-HSC expansion would be expected to improve survival of lethally irradiated recipients receiving limiting numbers of BMMC from PGE2 vs vehicle-treated mice. As predicted, recipients of BMMC from PGE2 treated mice had increased survival 30 days after transplantation compared to animals receiving BMMC from vehicle treated donors (150,000 donor cells: 80% vs 0% survival, p=0.0018; 75,000 donor cells: 53% vs 0% survival, p=0.0173). Taken together, these data demonstrate specific PGE2-dependent regulation of ST-HSC, and provide a unique and novel model to define control of HSC subsets. This finding implicates for the first time specialized regulation of HSC subsets. Moreover, these data indicate that selective therapeutic manipulation of ST-HSC could be exploited in clinical situations requiring rapid bone marrow reconstitution, such as in recovery from iatrogenic or pathologic myeloablative injury.

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Bone Marrow Homeostasis Is Impaired via JAK/STAT and Glucocorticoid Signaling in Cancer Cachexia Model

Cancers ◽

10.3390/cancers13051059 ◽

2021 ◽

Vol 13 (5) ◽

pp. 1059

Author(s):

Jinyeong Yu ◽

Sanghyuk Choi ◽

Aran Park ◽

Jungbeom Do ◽

Donghyun Nam ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Animal Model ◽

Bone Loss ◽

Cancer Cachexia ◽

Mononuclear Cells ◽

Bone Homeostasis ◽

Bone Marrow Mscs ◽

Hematopoietic Stem ◽

Glucocorticoid Signaling

Cancer cachexia is a multifactorial systemic inflammation disease caused by complex interactions between the tumor and host tissues via soluble factors. However, whether cancer cachexia affects the bone marrow, in particular the hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs), remains unclear. Here, we investigated the bone marrow and bone in a cancer cachexia animal model generated by transplanting Lewis lung carcinoma cells. The number of bone marrow mononuclear cells (BM-MNCs) started to significantly decrease in the cancer cachectic animal model prior to the discernable loss of muscle and fat. This decrease in BM-MNCs was associated with myeloid skewing in the circulation and the expansion of hematopoietic progenitors in the bone marrow. Bone loss occurred in the cancer cachexia animal model and accompanied the decrease in the bone marrow MSCs that play important roles in both supporting HSCs and maintaining bone homeostasis. Glucocorticoid signaling mediated the decrease in bone marrow MSCs in the cancer cachectic environment. The cancer cachexia environment also skewed the differentiation of the bone marrow MSCs toward adipogenic fate via JAK/STAT as well as glucocorticoid signaling. Our results suggest that the bone loss induced in cancer cachexia is associated with the depletion and the impaired differentiation capacity of the bone marrow MSCs.

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Altered colony-forming activities of bone marrow hematopoietic stem cells in mice following short-term in vivo exposure to parathion

The International Journal Of Cell Cloning ◽

10.1002/stem.5530050307 ◽

1987 ◽

Vol 5 (3) ◽

pp. 231-241 ◽

Cited By ~ 7

Author(s):

Vincent S. Gallicchio ◽

Thomas D. Watts ◽

George P. Casale ◽

Philip M. Bartholomew

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Hematopoietic Stem Cells ◽

Short Term ◽

Hematopoietic Stem

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Functional Resolution of Long-Term-and Short-Term- Hematopoietic Stem Cells

Journal of Stem Cell Research & Therapeutics ◽

10.15406/jsrt.2016.01.00044 ◽

2016 ◽

Vol 1 (6) ◽

Author(s):

Stephen H Bartelmez

Keyword(s):

Stem Cells ◽

Hematopoietic Stem Cells ◽

Short Term ◽

Hematopoietic Stem

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The active monomeric form of macrophage inflammatory protein-1 alpha interacts with high- and low-affinity classes of receptors on human hematopoietic cells

Blood ◽

10.1182/blood.v84.6.1790.1790 ◽

1994 ◽

Vol 84 (6) ◽

pp. 1790-1801 ◽

Cited By ~ 21

Author(s):

BR Avalos ◽

KJ Bartynski ◽

PJ Elder ◽

MS Kotur ◽

WG Burton ◽

...

Keyword(s):

Stem Cells ◽

Cell Cycle Progression ◽

Surface Proteins ◽

Biologically Active ◽

Polymorphonuclear Cells ◽

Monomeric Form ◽

Binding Studies ◽

Hematopoietic Stem ◽

Inflammatory Protein ◽

Macrophage Inflammatory Protein

Abstract Macrophage inflammatory protein-1 alpha (MIP-1 alpha) and its human homologue GOS19.1/LD78 are members of the C-C chemokine/intercrine family of secreted proteins. They have proinflammatory properties and also inhibit cell cycle progression of hematopoietic stem cells. Characterization of MIP-1 alpha receptor(s) has been confused because of its reported aggregation to inactive forms. Using a defined monomeric form of MIP-1 alpha that is biologically active for stem cell inhibition and induction of oxidative metabolism in polymorphonuclear cells, we report the detection of high- and low-affinity receptor classes on human leukemic CD34+ blast cells, promyelocytic cells, monocytes, peripheral blood neutrophils, and T cells. Both high- and low-affinity classes are expressed simultaneously in promyelocytes and neutrophils. The calculated kd for high-affinity receptors correlates with the concentrations of MIP-1 alpha required to induce a biologic effect on stem cells and neutrophils. Cross-linking studies show that MIP-1 alpha associates with two cell surface proteins with apparent molecular masses of 92 kD and 52 kD. Direct competition binding studies combined with studies on the inhibition of stem cells show that human and murine MIP-1 alpha have different receptor-binding and biologic properties.

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Detection and Isolation of Gene-Corrected Cells in Gaucher Disease Via a Fluorescence-Activated Cell Sorter Assay for Lysosomal Glucocerebrosidase Activity

Blood ◽

10.1182/blood.v89.9.3412 ◽

1997 ◽

Vol 89 (9) ◽

pp. 3412-3420 ◽

Cited By ~ 24

Author(s):

Matthew Lorincz ◽

Leonard A. Herzenberg ◽

Zhenjun Diwu ◽

John A. Barranger ◽

William G. Kerr

Keyword(s):

Stem Cells ◽

Gene Transfer ◽

Gaucher Disease ◽

Mononuclear Cells ◽

Disease Type ◽

Gene Correction ◽

Cell Sorter ◽

Hematopoietic Stem ◽

Gaucher Disease Type 1

Abstract Gaucher disease type 1 results from the accumulation of glucocerebroside in macrophages of the reticuloendothelial system, as a consequence of a deficiency in glucocerebrosidase (GC) activity. Recent improvements in the methodologies for introducing foreign genes into bone marrow stem cells have prompted several groups to test the efficacy of gene transfer therapy as a curative treatment for Gaucher disease. Limitations of this approach include the potential for insufficient engraftment of gene-corrected cells and incomplete transduction of hematopoietic stem cells using retroviral gene transfer. Overcoming these obstacles may be critical in the case of treatment for Gaucher disease type 1, because GC transduced cells have not been shown to have a growth advantage over noncorrected cells. Here, we describe the development and application of a novel, fluorescence-activated cell sorter based assay that directly quantitates GC activity at the single cell level. In a test of this application, fibroblasts from a Gaucher patient were transduced, and high expressing cells sorted based on GC activity. Reanalysis of cultured sorted fibroblasts reveals that these cells maintain high levels of enzymatic activity, compared with the heterogeneous population from which they were sorted. The assay is sufficiently sensitive to distinguish GC activity found in Gaucher patient monocytes from that in normal controls. Furthermore, preliminary results indicate that increased GC activity can be detected in transduced, CD34+ enriched peripheral blood mononuclear cells isolated from a Gaucher patient. This method should be a useful addition to current gene therapy protocols as a means to quantitatively assess gene correction of relevant cell populations and potentially purify transduced cells for transplantation.

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