Nitric Oxide Is a Regulator of Hematopoietic Stem Cell Activity

Michurina, Tatyana V.; Krasnov, Peter; Balazs, Alejandro B.; Nakaya, Naoki; Vasilieva, Tamara V.; Kuzin, Boris; Khrushchov, N. G.; Mulligan, Richard C.; Enikolopov, Grigori

doi:10.1016/j.ymthe.2004.05.030

Cited by 68 publications

(64 citation statements)

References 36 publications

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“…Particularly, nNOS expression was stable during maturation, iNOS consistently increased with maturation, whereas eNOS was more in the immature cells and was substantially reduced in the mature cells. As stromal cell derived NO has been shown to regulate hematopoiesis (32), studies are, however, required to explore the importance of NO derived from neutrophil precursor cells in hematopoiesis, which are highest in bone marrow and have the capability to produce NO efficiently. This study provides molecular characteristics of NOS isoforms in rat neutrophils starting from its origin in bone marrow to the mature peripheral PMNs.…”

Section: Discussionmentioning

confidence: 99%

Functional and molecular characterization of NOS isoforms in rat neutrophil precursor cells

et al. 2010

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Previous studies from this laboratory have demonstrated importance of neutrophilderived nitric oxide (NO) in free radical generation, characterized nitric oxide synthase (NOS) isoforms, and have reported subcellular distribution of NOS in rat peripheral neutrophils. Maximum number of neutrophils are added per day to the circulation from bone marrow, thus neutrophils might add substantial amount of NO in the bone marrow. NO generating ability and NOS isoforms characteristics in bone marrow neutrophil precursor cells is, however, still unexplored. This study was, therefore, undertaken to investigate NO generation ability and the molecular/biochemical characteristics of NOS isoforms in neutrophil precursor cells. The neutrophil precursors were separated on Percoll density gradient and characterized by Giemsa staining, CD markers, and by their size and granularity at various stages of maturation as Bands 1, 2, and 3. Mature neutrophils were efficient in free radical generation and phagocytosis, whereas immature cells had more mitochondria and myeloperoxidase. Amount of NO augmented from immature to mature neutrophils as assessed by fluorescent probe DAF-2DA and Griess reagent. Measurement of NOS enzyme activity further confirmed the functional status of NOS in these cells. NOS isoforms were differentially expressed during neutrophil maturation as confirmed by enzyme activity, Western blotting, flowcytometry, and RT-PCR. Expression of nNOS was predominantly stable in all the stages of neutrophil maturation. iNOS expression was, however, consistently augmented during maturation, whereas eNOS expression was downregulated with neutrophil maturation. Furthermore, all NOS isoforms proteins were distributed in cytosol as well as nucleus as assessed by confocal microscopy. This study for the first time report biochemical and molecular characteristics of NOS isoforms in rat neutrophil precursor cells. ' 2010 International Society for Advancement of Cytometry Key terms nitric oxide; NOS; neutrophils; neutrophil precursor cells; maturation NEUTROPHILS (PMNs) are orchestrating cells of the innate immune system (1,2), circulating through the body and extravasating to the sites of infection and injury, to perform important roles in the host defense. This process involves recruitment of lysosomes and various types of granules to release proteolytic enzymes, antimicrobial peptides, and free radical formation (1,2). PMN development in the bone marrow has classically been divided into six stages myeloblasts (MBs), promyelocytes (PMs), myelocytes (MCs), metamyelocytes (MMs), band cells (BCs), and segmented neutrophil on the basis of cell size, nuclear morphology, and granule content (3,4). Production of these cells is continuous to provide the continual demand for the tissues and mostly to maintain the circulating pool in the blood.Nitric oxide (NO), a versatile signaling molecule, mediates immune response, vasodilation, neurotransmission, proliferation, and apoptosis (5-8). NO is synthesized by a class of NADPH-dependent N...

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Section: Discussionmentioning

confidence: 99%

Functional and molecular characterization of NOS isoforms in rat neutrophil precursor cells

et al. 2010

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“…Moreover, in the transplantation model, this increase is followed by a transient increase in the number of neutrophils in the peripheral blood (18), indicating that modulation of NOS activity may be used for therapeutic intervention.…”

Section: Introductionmentioning

confidence: 99%

“…In the hematopoietic system, NO contributes to the regulation of hematopoietic stem and progenitor cells in the bone marrow; exposure of mice to NOS inhibitors, either directly or after irradiation and bone marrow transplantation, increases the number of stem and progenitor cells in the bone marrow (18). Moreover, in the transplantation model, this increase is followed by a transient increase in the number of neutrophils in the peripheral blood (18), indicating that modulation of NOS activity may be used for therapeutic intervention.…”

Section: Introductionmentioning

confidence: 99%

“…There are three NOS genes in the mammalian genome, coding for the neuronal, endothelial, and inducible isoforms of NOS (nNOS, eNOS, and iNOS, respectively), and mRNA of each NOS isoform can be reliably detected in mouse bone marrow (18). Furthermore, nNOS mRNA has been detected in neutrophils (19)(20)(21)(22), eNOS mRNA has been detected in lymphocytes, megakaryocytes, and platelets (19,23), and iNOS mRNA has been found in megakaryocytes, eosinophils, and unstimulated monocytes (21,24).…”

Section: Introductionmentioning

confidence: 99%

“…Although the action of NO in the hematopoietic system can be readily demonstrated (18,(25)(26)(27)(28)(29), neither the contribution of individual NOS isoforms nor their mode of action (autocrine vs. paracrine) is understood. We show here that NOS proteins are expressed in bone marrow stroma, that expression of the nNOS isoform strongly correlates with the ability of a panel of stromal cell lines to support hematopoietic stem and early progenitor cells in vitro, and that genetic inactivation of the nNOS gene increases the number of colonies that can be generated from bone marrow and spleen.…”

Section: Introductionmentioning

confidence: 99%

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Neuronal Nitric Oxide Synthase Contributes to the Regulation of Hematopoiesis

Krasnov

Michurina

Packer

et al. 2008

Mol Med

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Nitric oxide (NO) signaling is important for the regulation of hematopoiesis. However, the role of individual NO synthase (NOS) isoforms is unclear. Our results indicate that the neuronal NOS isoform (nNOS) regulates hematopoiesis in vitro and in vivo. nNOS is expressed in adult bone marrow and fetal liver and is enriched in stromal cells. There is a strong correlation between expression of nNOS in a panel of stromal cell lines established from bone marrow and fetal liver and the ability of these cell lines to support hematopoietic stem cells; furthermore, NO donor can further increase this ability. The number of colonies generated in vitro from the bone marrow and spleen of nNOS-null mutants is increased relative to wild-type or inducible-or endothelial NOS knockout mice. These results describe a new role for nNOS beyond its action in the brain and muscle and suggest a model where nNOS, expressed in stromal cells, produces NO which acts as a paracrine regulator of hematopoietic stem cells.

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Expression of inducible NOS is indispensable for the antiproliferative and proapoptotic effect of imatinib in BCR–ABL positive cells

Singh

Awasthi

Dubey

et al. 2021

Journal of Leukocyte Biology

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Chronic myeloid leukemia (CML) is characterized by constitutive BCR–ABL kinase activity, an aggressive proliferation of immature cells, and reduced differentiation. Targeting tyrosine kinase activity of BCR–ABL with imatinib is an effective therapy for the newly diagnosed CML patients; however, 20%–30% of the patients initially treated with imatinib eventually experience treatment failure. Therefore, early identification of these patients is of high clinical relevance. In the present study, we by undertaking a direct comparison of inducible NOS (iNOS) status in neutrophils from healthy volunteers, newly diagnosed, imatinib responder, and resistant CML patients as well as by conducting in vitro studies in K562 cells demonstrated that inhibition of BCR–ABL by imatinib or siRNA significantly enhanced NO generation and iNOS expression. Indeed, patients exhibiting treatment failure or imatinib resistance were less likely to induce NO generation/iNOS expression. Our findings further demonstrated that imatinib mediated antiproliferative and proapoptotic effect in BCR–ABL+ cells associated with enhanced iNOS expression, and it was significantly prevented in the presence of L‐NAME, 1400W, or iNOS siRNA. Overexpression of iNOS in K562 cells expectedly enhanced imatinib sensitivity on cytostasis and apoptosis, even at lower concentration (0.1 μM) of imatinib. Mechanistically, imatinib or BCR–ABL siRNA following deglutathionylation of NF‐κB, enhanced its binding to iNOS promoter and induced iNOS transcription. Deglutathionylation of procaspase‐3 however associated with increased caspase‐3 activity and cell apoptosis. Taken together, results obtained suggest that monitoring NO/iNOS level could be useful to identify patients likely to be responsive or resistant to imatinib and can be used to personalized alternative therapy.

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Nitric Oxide Is a Regulator of Hematopoietic Stem Cell Activity

Cited by 68 publications

References 36 publications

Functional and molecular characterization of NOS isoforms in rat neutrophil precursor cells

Functional and molecular characterization of NOS isoforms in rat neutrophil precursor cells

Neuronal Nitric Oxide Synthase Contributes to the Regulation of Hematopoiesis

Expression of inducible NOS is indispensable for the antiproliferative and proapoptotic effect of imatinib in BCR–ABL positive cells

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