Nitric Oxide-Induced Murine Hematopoietic Stem Cell Fate Involves Multiple Signaling Proteins, Gene Expression, and Redox Modulation

Abstract

There are a growing number of reports showing the influence of redox modulation in cellular signaling. Although the regulation of hematopoiesis by reactive oxygen species (ROS) and reactive nitrogen species (RNS) has been described, their direct participation in the differentiation of hematopoietic stem cells (HSCs) remains unclear. In this work, the direct role of nitric oxide (NO • ), a RNS, in the modulation of hematopoiesis was investigated using two sources of NO•

M1 and M2 macrophages differentially regulate hematopoietic stem cell self-renewal and ex vivo expansion

“…This finding is in agreement with the previous observations that NO can inhibit HSC self-renewal by stimulating HSC proliferation and differentiation and induction of HSC and HPC apoptosis. [23][24][25] Promotion of HSC selfrenewal and expansion by mouse M2-MFs is mediated, in part, by Arg1, because inhibition of Arg1 activity with amino-2-borono-6-hexanoic acid or knockout of Arg1 in purified MFs attenuated the effect of M2-MFs on HSC self-renewal. The mechanism by which Arg1 from M2-MFs regulates HSC self-renewal may be attributable, at least in part, to the production of spermidine, because spermidine, but not L-ornithine and spermine, could partially replace M2-MFs to increase the expansion of mouse BM HSCs.…”

“…Potentially, NO derived from NOS2 can promote HSC and hematopoietic progenitor cell (HPC) proliferation and differentiation at the expense of HSC self-renewal. 23,24 In addition, exposure to NO induces HSC and HPC apoptosis. 25 In contrast, polyamines, such as spermidine synthesized through the arginine metabolic pathway initiated by Arg1, have the potential to promote HSC self-renewal via stimulation of autophagy.…”

Regulation of Cellular Redox Signaling by Matricellular Proteins in Vascular Biology, Immunology, and Cancer

“…In vitro, treatment with the NO donor linsidomine (3-morpholinosydnonimine, SIN-1) stimulated proliferation of muscle satellite cells (a precursor muscle cell type), and this was inhibited by concurrent treatment with L-NAME (20). Hematopoietic stem cells treated with an NO donor proliferated faster and had increased expression of the self-renewal transcription factor cMyc (183). Thus, multiple stem cell populations are sensitive to the proliferative/cell cycle stimulating effects of NO.…”

Reliability of ROS and RNS detection in hematopoietic stem cells − potential issues with probes and target cell population

“…Hydroethidine (HE) is commonly used for the detection of intracellular O 2 • − in studies of HSCs and HPCs (Jung et al, 2013;Nogueira-Pedro et al, 2014;Tesio et al, 2011;Wang et al, 2010). Although O 2 • − can directly oxidize HE to its radical, the reaction is relatively slow and is more likely to be performed by other cellular oxidants present (Winterbourn, 2014 the non-specific reaction, any specificity of the fluorescent signal for O 2 • − needs to be confirmed with ROS-scavenging enzymes and NOS inhibitors (Zielonka et al, 2012b).…”

“…Furthermore, inhibition of nitric oxide synthase (NOS) increases the number of stem cells in the BM and the longevity of hematopoiesis in continuous BM cultures (Epperly et al, 2007;Michurina et al, 2004). However, compared with the work on ROS, there are considerably fewer studies that address RNS-mediated effects on HSCs and the signaling networks they participate in (Aleksinskaya et al, 2013;Bonafèet al, 2015;Nogueira-Pedro et al, 2014).…”