Ras, Rac1, and phosphatidylinositol-3-kinase (PI3K) signaling in nitric oxide induced endothelial cell migration

Borges, Roberta Eller; Batista, Wagner L.; Costa, P.; Tokikawa, Rita; Curcio, Marli F.; Strumillo, Scheilla Teixeira; Sartori, Adriano; Moraes, Miriam S.; Oliveira, Gustavo H. C.; Taha, Murched Omar; Fonseca, Fabiana Valéria da; Stern, Arnold; Monteiro, Hugo P.

doi:10.1016/j.niox.2015.03.004

Cited by 19 publications

(12 citation statements)

References 48 publications

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“…7 ) can increase cell motility compared to controls, and the effect is diminished by L-NAME treatment, suggesting that NO plays a role in regulating rBMSC cell migration (Fig. 7 ) which has been previously demonstrated for endothelial cell migration [ 61 ]. Together, these findings show that genetic manipulation of MSCs to enhance bioavailable NO may upregulate VEGF-A/PDGFRα and FGF2/FGFR2 signalling pathways to promote angiogenesis (Fig.…”

Section: Discussionsupporting

confidence: 61%

Minicircle DNA-mediated endothelial nitric oxide synthase gene transfer enhances angiogenic responses of bone marrow-derived mesenchymal stem cells

et al. 2016

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BackgroundNon-viral-based gene modification of adult stem cells with endothelial nitric oxide synthase (eNOS) may enhance production of nitric oxide and promote angiogenesis. Nitric oxide (NO) derived from endothelial cells is a pleiotropic diffusible gas with positive effects on maintaining vascular tone and promoting wound healing and angiogenesis. Adult stem cells may enhance angiogenesis through expression of bioactive molecules, and their genetic modification to express eNOS may promote NO production and subsequent cellular responses.MethodsRat bone marrow-derived mesenchymal stem cells (rBMSCs) were transfected with a minicircle DNA vector expressing either green fluorescent protein (GFP) or eNOS. Transfected cells were analysed for eNOS expression and NO production and for their ability to form in vitro capillary tubules and cell migration. Transcriptional activity of angiogenesis-associated genes, CD31, VEGF-A, PDGFRα, FGF2, and FGFR2, were analysed by quantitative polymerase chain reaction.ResultsMinicircle vectors expressing GFP (MC-GFP) were used to transfect HEK293T cells and rBMSCs, and were compared to a larger parental vector (P-GFP). MC-GFP showed significantly higher transfection in HEK293T cells (55.51 ± 3.3 %) and in rBMSC (18.65 ± 1.05 %) compared to P-GFP in HEK293T cells (43.4 ± 4.9 %) and rBMSC (15.21 ± 0.22 %). MC-eNOS vectors showed higher transfection efficiency (21 ± 3 %) compared to P-eNOS (9 ± 1 %) and also generated higher NO levels. In vitro capillary tubule formation assays showed both MC-eNOS and P-eNOS gene-modified rBMSCs formed longer (14.66 ± 0.55 mm and 13.58 ± 0.68 mm, respectively) and a greater number of tubules (56.33 ± 3.51 and 51 ± 4, respectively) compared to controls, which was reduced with the NOS inhibitor L-NAME. In an in vitro wound healing assay, MC-eNOS transfected cells showed greater migration which was also reversed by L-NAME treatment. Finally, gene expression analysis in MC-eNOS transfected cells showed significant upregulation of the endothelial-specific marker CD31 and enhanced expression of VEGFA and FGF-2 and their corresponding receptors PDGFRα and FGFR2, respectively.ConclusionsA novel eNOS-expressing minicircle vector can efficiently transfect rBMSCs and produce sufficient NO to enhance in vitro models of capillary formation and cell migration with an accompanying upregulation of CD31, angiogenic growth factor, and receptor gene expression.

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Section: Discussionsupporting

confidence: 61%

Minicircle DNA-mediated endothelial nitric oxide synthase gene transfer enhances angiogenic responses of bone marrow-derived mesenchymal stem cells

et al. 2016

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“…Cyclic GMP in turn (i) activates cGMP-dependent protein kinases (PKG), (ii) activates or inhibits cAMP-specific phosphodiesterases (PDEs), and (iii) opens cyclic nucleotide-gated cation channels, leading to a wide range of context-specific physiological and developmental outcomes in animals, some of which include ERK signalling2941777879. In addition, NO can impact on cell state directly by S-nitrosylation of cysteine residues of signalling pathway components and transcription factors438081, including members of the Ras-GTPase super family8283. Thus it is possible that ERK activation may occur via the S-nitrosylation of Ras, or an alternative mechanism, in A. queenslandica (Fig.…”

Section: Discussionmentioning

confidence: 99%

An ancient role for nitric oxide in regulating the animal pelagobenthic life cycle: evidence from a marine sponge

Ueda

Richards

Degnan

et al. 2016

Sci Rep

111

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In many marine invertebrates, larval metamorphosis is induced by environmental cues that activate sensory receptors and signalling pathways. Nitric oxide (NO) is a gaseous signalling molecule that regulates metamorphosis in diverse bilaterians. In most cases NO inhibits or represses this process, although it functions as an activator in some species. Here we demonstrate that NO positively regulates metamorphosis in the poriferan Amphimedon queenslandica. High rates of A. queenslandica metamorphosis normally induced by a coralline alga are inhibited by an inhibitor of nitric oxide synthase (NOS) and by a NO scavenger. Consistent with this, an artificial donor of NO induces metamorphosis even in the absence of the alga. Inhibition of the ERK signalling pathway prevents metamorphosis in concert with, or downstream of, NO signalling; a NO donor cannot override the ERK inhibitor. NOS gene expression is activated late in embryogenesis and in larvae, and is enriched in specific epithelial and subepithelial cell types, including a putative sensory cell, the globular cell; DAF-FM staining supports these cells being primary sources of NO. Together, these results are consistent with NO playing an activating role in induction of A. queenslandica metamorphosis, evidence of its highly conserved regulatory role in metamorphosis throughout the Metazoa.

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“…A recent population-based cohort study conducted in Taiwan has shown a decreased risk of diabetic retinopathy, and its progression to vision-threatening diabetic retinopathy in Taiwanese patients receiving statin therapy [94]; additional studies are needed to ascertain the link between statins and diabetic retinopathy. Furthermore, since nitric oxide can also activate Rac1 [95], several nitric oxide releasing molecules have been analyzed for their protective effect against oxidative stress, and VP10/39 (caffeic acid phenethyl ester derivative) has been shown to protect retinal pigment epithelial cells against oxidative stress [96]; however, the effect of such molecules on diabetic retinopathy remains to be investigated.…”

Section: Therapeutic Targetsmentioning

confidence: 99%

The Regulatory Role of Rac1, a Small Molecular Weight GTPase, in the Development of Diabetic Retinopathy

Sahajpal

Kowluru

2019

JCM

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Diabetic retinopathy, a microvascular complication of diabetes, remains the leading cause of vision loss in working age adults. Hyperglycemia is considered as the main instigator for its development, around which other molecular pathways orchestrate. Of these multiple pathways, oxidative stress induces many metabolic, functional and structural changes in the retinal cells, leading to the development of pathological features characteristic of this blinding disease. An increase in cytosolic reactive oxygen species (ROS), produced by cytosolic NADPH oxidase 2 (Nox2), is an early event in the pathogenesis of diabetic retinopathy, which leads to mitochondrial damage and retinal capillary cell apoptosis. Activation of Nox2 is mediated through an obligatory small molecular weight GTPase, Ras-related C3 botulinum toxin substrate 1 (Rac1), and subcellular localization of Rac1 and its activation are regulated by several regulators, rendering it a complex biological process. In diabetes, Rac1 is functionally activated in the retina and its vasculature, and, via Nox2-ROS, contributes to mitochondrial damage and the development of retinopathy. In addition, Rac1 is also transcriptionally activated, and epigenetic modifications play a major role in this transcriptional activation. This review focusses on the role of Rac1 and its regulation in the development and progression of diabetic retinopathy, and discusses some possible avenues for therapeutic interventions.

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Ras, Rac1, and phosphatidylinositol-3-kinase (PI3K) signaling in nitric oxide induced endothelial cell migration

Cited by 19 publications

References 48 publications

Minicircle DNA-mediated endothelial nitric oxide synthase gene transfer enhances angiogenic responses of bone marrow-derived mesenchymal stem cells

Minicircle DNA-mediated endothelial nitric oxide synthase gene transfer enhances angiogenic responses of bone marrow-derived mesenchymal stem cells

An ancient role for nitric oxide in regulating the animal pelagobenthic life cycle: evidence from a marine sponge

The Regulatory Role of Rac1, a Small Molecular Weight GTPase, in the Development of Diabetic Retinopathy

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