The UDP-sugar-sensing P2Y14receptor promotes Rho-mediated signaling and chemotaxis in human neutrophils

The Gi-coupled P2Y14receptor (P2Y14-R) is potently activated by UDP-sugars and UDP. Although P2Y14-R mRNA is prominently expressed in circulating neutrophils, the signaling pathways and functional responses associated with this receptor are undefined. In this study, we illustrate that incubation of isolated human neutrophils with UDP-glucose resulted in cytoskeleton rearrangement, change of cell shape, and enhanced cell migration. We also demonstrate that UDP-glucose promotes rapid, robust, and concentration-dependent activation of RhoA in these cells. Ecto-nucleotidases expressed on neutrophils rapidly hydrolyzed extracellular ATP, but incubation with UDP-glucose for up to 1 h resulted in negligible metabolism of the nucleotide-sugar. HL60 human promyelocytic leukemia cells do not express the P2Y14-R, but neutrophil differentiation of HL60 cells with DMSO resulted in markedly enhanced P2Y14-R expression. Accordingly, UDP-glucose, UDP-galactose, and UDP- N-acetylglucosamine promoted Rho activation in differentiated but not in undifferentiated HL60 cells. Stable expression of recombinant human P2Y14-R conferred UDP-sugar-promoted responses to undifferentiated HL60 cells. UDP-glucose-promoted RhoA activation also was accompanied by enhanced cell migration in differentiated HL60 cells, and these responses were blocked by Rho kinase inhibitors. These results support the notion that UDP-glucose is a stable and potent proinflammatory mediator that promotes P2Y14-R-mediated neutrophil motility via Rho/Rho kinase activation.

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Faculty Opinions recommendation of Rho kinase inhibitors block melanoma cell migration and inhibit metastasis.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.725421408.793516995 ◽

2016 ◽

Author(s):

Victoria Sanz-Moreno

Keyword(s):

Cell Migration ◽

Melanoma Cell ◽

Kinase Inhibitors ◽

Rho Kinase ◽

Rho Kinase Inhibitors

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Investigation of the inhibitory effects of HA-1077 and Y-32885 on the translocation of PKCβI, PKCβII and PKCζ in human neutrophils

Mediators of Inflammation ◽

10.1080/09629350120102334 ◽

2001 ◽

Vol 10 (6) ◽

pp. 315-321 ◽

Cited By ~ 3

Author(s):

Xavier Siomboing ◽

Bernard Gressier ◽

Thierry Dine ◽

Claude Brunet ◽

Michel Luyckx ◽

...

Keyword(s):

Kinase Inhibitors ◽

Rho Kinase ◽

Polymorphonuclear Neutrophils ◽

Human Neutrophils ◽

Inhibitory Effects ◽

Rho Kinase Inhibitors ◽

Kinase C ◽

Compound Synthesis ◽

Plasmic Membrane ◽

Specific Ligand

To transmit the information inside the cell, one possibility is the action of an enzyme called kinase that phosphorylates other proteins. To study these enzymes, chemical compound synthesis was needed to know the function and the mechanism of activation. The major difficulty is creating a specific molecule for one kinase. In this study, we test the action of Rho-kinase inhibitors (HA-1077 and Y-32885) on protein kinase C (PKC) in the respiratory burst in the human polymorphonuclear neutrophils. We have shown that these compounds could inhibit the anion superoxide production. To prove their action on PKC, we have shown a decrease of binding of a specific ligand (phorbol-12,13-dibutyrate) with each inhibitor. During its activation, PKC was translocated from the cytoplasm to the plasmic membrane. We have also shown an inhibition of this translocation, proving an inhibition of PKC by HA-1077 and Y-32885.

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Rho Kinase Inhibitors Block Melanoma Cell Migration and Inhibit Metastasis

Cancer Research ◽

10.1158/0008-5472.can-14-2156 ◽

2015 ◽

Vol 75 (11) ◽

pp. 2272-2284 ◽

Cited By ~ 68

Author(s):

Amine Sadok ◽

Afshan McCarthy ◽

John Caldwell ◽

Ian Collins ◽

Michelle D. Garrett ◽

...

Keyword(s):

Cell Migration ◽

Melanoma Cell ◽

Kinase Inhibitors ◽

Rho Kinase ◽

Rho Kinase Inhibitors

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Matrix-independent stimulation of human tubular epithelial cell migration by Rho kinase inhibitors

Journal of Cellular Physiology ◽

10.1002/jcp.22079 ◽

2010 ◽

pp. n/a-n/a

Author(s):

Sven Kroening ◽

Jana Stix ◽

Christof Keller ◽

Cedric Streiff ◽

Margarete Goppelt-Struebe

Keyword(s):

Cell Migration ◽

Epithelial Cell ◽

Kinase Inhibitors ◽

Rho Kinase ◽

Tubular Epithelial Cell ◽

Rho Kinase Inhibitors ◽

Epithelial Cell Migration ◽

Stimulation Of

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Rho kinase activation and ROS production contributes to the cooling enhanced contraction in cutaneous equine digital veins

Journal of Applied Physiology ◽

10.1152/japplphysiol.01301.2009 ◽

2010 ◽

Vol 109 (1) ◽

pp. 11-18 ◽

Cited By ~ 5

Author(s):

H. Zerpa ◽

Y. Berhane ◽

H. Woodcock ◽

J. Elliott ◽

S. R. Bailey

Keyword(s):

Kinase Inhibitors ◽

Contractile Response ◽

Rho Kinase ◽

Maximum Response ◽

Cellular Mechanisms ◽

Kinase Activation ◽

Rho Kinase Inhibitors ◽

Adrenoceptor Agonist ◽

Control Response ◽

Adrenoceptor Agonists

A decrease in environmental temperature can directly affect the contractility of cutaneous vasculature, mediated in part by α2-adrenoceptors. Most of the cellular mechanisms underlying the cooling-enhanced contractility to α2-adrenoceptor agonists have been reported in cutaneous arteries but little information is available on cutaneous veins. To investigate the cellular mechanisms associated with the cooling-enhanced contraction to UK-14304 (α2-adrenoceptor agonist), isolated equine digital veins (EDVs) were studied at 30°C and 22°C. The effects of inhibitors were studied on the contractile response to UK-14304 (0.1 μM). The cooling-enhanced responses were inhibited by Rho kinase inhibitors [maximum response to UK-14304 95.2 ± 8% of response to depolarizing Krebs solution (DKS) in control vessels cooled to 22°C, compared with 31.4 ± 6% in the presence of fasudil 1 μM and 75.8 ± 6% with Y-27632 0.1 μM] and the effects of these inhibitors were considerably less at 30°C (control response 56.4 ± 5% of DKS; 34.9 ± 6% with fasudil 1 μM and 50.6 ± 9% with Y-27632 0.1 μM). Furthermore, Western blotting showed that one of the downstream targets for Rho kinase activity, ezrin/radixin/moesin, was phosphorylated after cooling and reduced by fasudil (1 μM) only at 22°C. The activation of protein kinase C contributed to the contractile response, but predominantly at 30°C (maximum response 82.3 ± 9% of DKS for control; 57.7 ± 10% in the presence of chelerythrine 10 μM) with no significant effect at 22°C. The reduction of the response at 22°C by antioxidants, rotenone (14% reduction), and tempol (21% reduction) suggested the contribution of reactive oxygen species (ROS). No evidence was obtained to support the participation of tyrosine kinase. These data demonstrate that Rho kinase activation and the production of ROS contributes to the cooling-enhanced contraction in these cutaneous digital veins.

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Probing Cellular Outcomes Using Heterobivalent Constructs

10.26434/chemrxiv.7613213.v1 ◽

2019 ◽

Author(s):

Rohit Bhadoria ◽

Kefeng Ping ◽

Christer Lohk ◽

Ivar Järving ◽

Pavel Starkov

Keyword(s):

Cell Viability ◽

Small Molecules ◽

Cellular Uptake ◽

Kinase Inhibitors ◽

Rho Kinase ◽

Intracellular Delivery ◽

Rho Kinase Inhibitors

<div> <div> <div> <p>Conjugation techniques are central to improving intracellular delivery of bioactive small molecules. However, tracking and assessing the overall biological outcome of these constructs remains poorly understood. We addressed this issue by having developed a focused library of heterobivalent constructs based on Rho kinase inhibitors to probe various scenarios. By comparing induction of a phenotype of interest vs. cell viability vs. cellular uptake, we demonstrate that such conjugates indeed lead to divergent cellular outcomes. </p> </div> </div> </div>

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Rho Kinase Inhibitors: Potential Treatments for Diabetes and Diabetic Complications

Current Pharmaceutical Design ◽

10.2174/138161212800672688 ◽

2012 ◽

Vol 18 (20) ◽

pp. 2964-2973 ◽

Cited By ~ 16

Author(s):

Hong Zhou ◽

Yong-jun Li

Keyword(s):

Diabetic Complications ◽

Kinase Inhibitors ◽

Rho Kinase ◽

Rho Kinase Inhibitors

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Neovascular Macular Degeneration: A Review of Etiology, Risk Factors, and Recent Advances in Research and Therapy

International Journal of Molecular Sciences ◽

10.3390/ijms22031170 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1170

Author(s):

Arunbalaji Pugazhendhi ◽

Margaret Hubbell ◽

Pooja Jairam ◽

Balamurali Ambati

Keyword(s):

Risk Factors ◽

Macular Degeneration ◽

Kinase Inhibitors ◽

Rho Kinase ◽

The Elderly ◽

Age Related Macular Degeneration ◽

Gene Therapies ◽

Rho Kinase Inhibitors ◽

Age Related ◽

Endothelial Growth Factor

Neovascular age-related macular degeneration (exudative or wet AMD) is a prevalent, progressive retinal degenerative macular disease that is characterized by neovascularization of the choroid, mainly affecting the elderly population causing gradual vision impairment. Risk factors such as age, race, genetics, iris color, smoking, drinking, BMI, and diet all play a part in nvAMD’s progression, with anti-vascular endothelial growth factor (anti-VEGF) therapy being the mainstay of treatment. Current therapeutic advancements slow the progression of the disease but do not cure or reverse its course. Newer therapies such as gene therapies, Rho-kinase inhibitors, and levodopa offer potential new targets for treatment.

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