Stem Cell Factor Improves the Repopulating Ability of Primitive Hematopoietic Stem Cells after Sublethal Irradiation (and, to a Lesser Extent) after Bone Marrow Transplantation in Mice

Gardner, Renée V.; Oliver, Peter; Astle, Clinton M.

doi:10.1002/stem.160112

Cited by 12 publications

(11 citation statements)

References 44 publications

(47 reference statements)

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“…Because of our greater capacity to quantitate stem cell numbers in experimental animal models, our understanding of this discrepancy is best exemplified by several genetic defects of mice, including the W and Sl mutants, 21 mice given myelosuppressive or myeloablative radiation therapy, 22 cats administered busulfan, 23 and mice nullizygous for the Mpl protooncogene. 11 In humans, children with congenital amegakaryocytic thrombocytopenia and patients soon after HSC transplantation also represent states in which stem and progenitor cell numbers are significantly reduced 10,24 but all or most of the peripheral blood cell counts are normal.…”

Section: Discussionmentioning

confidence: 99%

Lineage-specific growth factors can compensate for stem and progenitor cell deficiencies at the postprogenitor cell level: an analysis of doubly TPO- and G-CSF receptor–deficient mice

Kaushansky

Fox

Lin

et al. 2002

Blood

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Multiple lines of evidence indicate that thrombopoietin (TPO) substantially impacts the number of hematopoietic stem cells and progenitors of all myeloid lineages. Nevertheless, tpo knock-out mice (T ؊ ) display thrombocytopenia only; blood erythroid and neutrophil levels are normal despite 60% to 85% reductions in stem and progenitor cells. The compensatory mechanism(s) for these deficiencies remains uncertain; lineage-specific cytokines such as erythropoietin or granulocyte colony-stimulating factor (G-CSF) have been postulated but never proven to be responsible. To directly test whether G-CSF can compensate for the myeloid progenitor cell reduction in the T ؊ model of hematopoietic deficiency, T ؊ and G-CSF-receptor knock-out (GR ؊ ) mice were crossed, and F1 animals bred to obtain doubly nullizygous mice (T ؊ GR ؊ ). This experiment also allowed us to test the hypothesis that G-CSF contributes to the residual platelet production in T ؊ mice. We found that T ؊ GR ؊ F2 mice displayed similar blood platelet levels as that seen in T ؊ mice, indicating that G-CSF does not account for the residual megakaryopoiesis in T ؊ mice. However, we also noted excessive perinatal mortality of T ؊ GR ؊ animals, caused by infection due to a profound and significant decrease in marrow and peripheral blood neutrophils, far greater than that seen in either T ؊ or GR ؊ mice. These data indicate that in the additional absence of GR, T ؊ mice cannot compensate for their 62% reduction in myeloid progenitors and become profoundly neutropenic, supporting the hypothesis that G-CSF can compensate for the myeloid effects of TPO deficiency by expanding the pool of cells between the granulocyte-macrophage colony-forming unit and mature neutrophil stages of granulopoiesis. IntroductionA major step in our understanding of the regulation of blood cell production came with the cloning and characterization of hematopoietic cytokines and their receptors. Based on their activities in various in vitro assays and administration in vivo, the lineage specificity of these glycoprotein hormones was quickly determined. At present, the hematopoietic cytokines can be broadly divided into those that primarily affect the early aspects of hematopoietic development, working on multiple cell types with multilineage potential, and a subfamily of lineage-dominant cytokines that work primarily on cells committed to a single hematopoietic lineage. 1 It is also clear that the most profound hematopoietic effects in vivo are exerted by the lineage-dominant cytokines erythropoietin (EPO), granulocyte colony-stimulating factor (G-CSF), and thrombopoietin (TPO).There is little doubt that G-CSF is the primary regulator of steady state and reactive neutrophil production. 2 The administration of the cytokine to normal animals leads to profound increases in neutrophil levels, and mice in which the G-CSF or G-CSFreceptor (G-CSF-R) genes have been genetically eliminated are reported to have neutrophil counts about 25% that of littermate controls. 3,4 Likewise, the use of TPO ...

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Section: Discussionmentioning

confidence: 99%

Lineage-specific growth factors can compensate for stem and progenitor cell deficiencies at the postprogenitor cell level: an analysis of doubly TPO- and G-CSF receptor–deficient mice

Kaushansky

Fox

Lin

et al. 2002

Blood

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“…Treatment with SCF induces HSPC expansion, radioprotection and subsequent repopulation of hematopoietic lineages, leading to improved recovery and survival after irradiation (Gardner et al, 1998; Tong et al, 1993; Zsebo et al, 1992). However, pleiotropic SCF actions also play critical roles in mast cell development, maturation and activation (Galli et al, 1993; Taylor et al, 1995).…”

Section: Introductionmentioning

confidence: 99%

Decoupling the Functional Pleiotropy of Stem Cell Factor by Tuning c-Kit Signaling

Chhabra

Starkl

et al. 2017

Cell

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SUMMARY Most secreted growth factors and cytokines are functionally pleiotropic because their receptors are expressed on diverse cell types. While important for normal mammalian physiology, pleiotropy limits the efficacy of cytokines and growth factors as therapeutics. Stem cell factor (SCF) is a growth factor that acts through the c-Kit receptor tyrosine kinase to elicit hematopoietic progenitor expansion, but can be toxic when administered in vivo because it concurrently activates mast cells. We engineered a mechanism-based SCF partial agonist that impaired c-Kit dimerization, truncating downstream signaling amplitude. This SCF variant elicited biased activation of hematopoietic progenitors over mast cells in vitro and in vivo. Mouse models of SCF-mediated anaphylaxis, radioprotection, and hematopoietic expansion revealed that this SCF partial agonist retained therapeutic efficacy while exhibiting virtually no anaphylactic off-target effects. The approach of biasing cell activation by tuning signaling thresholds and outputs has applications to many dimeric receptor-ligand systems.

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“…Adult stem and progenitor cells or cancer stem cells incorporating either of the two MDR transporters can be isolated by flow cytometry without significant impact in their physiological or biological condition. Actual verification that JC1 low cells has in vivo regenerative capacity in a test such as hematopoietic bone marrow repopulating capacity [35] is pending.…”

Section: Discussionmentioning

confidence: 99%

Application of JC1 for non-toxic isolation of cells with MDR transporter activity by flow cytometry

2017

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The DNA intercalating dye Hoechst 33342 or its close analog DCV are actively removed from cells by the multidrug resistance transporter ABCG2, a protein overexpressed in metastatic cells and somatic stem cells. In bivariate blue-red flow cytometry fluorescent plots active Hoechst or DCV efflux combined with a concentration dependent bathochromic shifts of these nuclear dyes leads to the segregation of the transporter-rich cells into a distinct cell cohort tilted towards the shorter wavelength axis of the plot, the cohort is generically known as the side population (SP). This feature has facilitated the surface marker-independent isolation of live stem cells. A drawback, though, is the known toxicity of Hoechst dyes. In this study we show that JC1, a bathochromic mitochondrial membrane potential-sensitive dye applied at proper concentration, can yield flow cytometry fluorescent emission bivariate plots containing a low JC1 accumulation (JC1low) cohort. Using a combination of multiple cell lines, ABC-transporter inhibitors and viral vector-driven insertion of the ABCG2 gene or ABCG2 and ABCB1 shRNAs we demonstrate that JC1low can be generated by either of the two aforementioned multidrug resistance transporters. Complete wash out of mitochondrial bound JC1 required more than 24 h. In spite of this tight binding, the dye did not affect either the mitochondrial membrane potentials or the proliferation rate. In contrast, contemporaneous with its nuclear accumulation, Hoechst 33342 or DVC, caused changes in the fluorescent emission of mitochondrial membrane potential sensitive dyes resembling the effects caused by the mitochondrial uncoupler FCCP. In a number of cell lines exposure to Hoechst resulted in marked slow-down of proliferation and abolition of ABCG2 transport activity during the subsequent 2 days but in K562 cells the exposure induced cell extended death. Overall, its lack of toxicity vis. a vis. the toxicity and genotoxicity of the DNA intercalating dyes makes JC1 an ideal tool for isolating live cells expressing high multidrug resistance transport activity.

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Stem Cell Factor Improves the Repopulating Ability of Primitive Hematopoietic Stem Cells after Sublethal Irradiation (and, to a Lesser Extent) after Bone Marrow Transplantation in Mice

Cited by 12 publications

References 44 publications

Lineage-specific growth factors can compensate for stem and progenitor cell deficiencies at the postprogenitor cell level: an analysis of doubly TPO- and G-CSF receptor–deficient mice

Lineage-specific growth factors can compensate for stem and progenitor cell deficiencies at the postprogenitor cell level: an analysis of doubly TPO- and G-CSF receptor–deficient mice

Decoupling the Functional Pleiotropy of Stem Cell Factor by Tuning c-Kit Signaling

Application of JC1 for non-toxic isolation of cells with MDR transporter activity by flow cytometry

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