Cross Talk between the Akt and p38α Pathways in Macrophages Downstream of Toll-Like Receptor Signaling

McGuire, Victoria A.; Gray, Alexander; Monk, Claire E.; Santos, Susana G.; Lee, Keunwook; Aubareda, Anna; Crowe, Jennifer; Ronkina, Natalia; Schwermann, Jessica; Batty, I H; Leslie, Nicholas R.; Dean, Jonathan L. E.; O’Keefe, Stephen J.; Boothby, Mark; Gaestel, Matthias; Arthur, J. Simon C.

doi:10.1128/mcb.01691-12

Cited by 67 publications

(72 citation statements)

References 76 publications

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“…Primary bone marrow derived macrophages (BMDMs) were prepared as described. 53 Cells were maintained on bacterial grade DMEM supplemented with 10% heat-inactivated fetal bovine serum, 2 mM L-glutamine, 100 U/ml of penicillin G, 100 μg/ml of streptomycin, 0.25 μg/ml of amphotericin (Invitrogen), and 5 ng/ml of macrophage colony-stimulating factor (M-CSF; PreProTech) for 7 days. Cells were then replated onto tissue culture grade plastic and used the following day.…”

Section: Methodsmentioning

confidence: 99%

Development of Selective Covalent Janus Kinase 3 Inhibitors

et al. 2015

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The Janus Kinases (JAKs) and their downstream effectors Signal Transducer and Activator of Transcription proteins (STATs) form a critical immune cell signaling circuit, which is of fundamental importance in innate immunity, inflammation and hematopoiesis and dysregulation is frequently observed in immune disease and cancer. The high degree of structural conservation of the JAK ATP binding pockets has posed a considerable challenge to medicinal chemists seeking to develop highly selective inhibitors as pharmacological probes and as clinical drugs. Here we report the discovery and optimization of 2,4-substituted pyrimidines as covalent JAK3 inhibitors that exploit a unique cysteine (Cys909) residue in JAK3. Investigation of structure-activity-relationship (SAR) utilizing biochemical and transformed Ba/F3 cellular assays resulted in identification of potent and selective inhibitors such as compounds 9 and 45. A 2.9 Å co-crystal structure of JAK3 in complex with 9 confirms the covalent interaction. Compound 9 exhibited decent pharmacokinetic properties and is suitable for use in vivo. These inhibitors provide a set of useful tools to pharmacologically interrogate JAK3-dependent biology.

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Section: Methodsmentioning

confidence: 99%

Development of Selective Covalent Janus Kinase 3 Inhibitors

et al. 2015

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“…SAPK/JNK and p38 activation by LPS promotes the synthesis and release of pro-inflammatory mediators by macrophages [18]. In addition, p38 activates mitogen-activated protein kinase-activated protein kinase 2 (MK2), which indirectly regulates Akt activity by controlling phosphatidylinositol 3, 4,5-trisphosphate (PIP 3 ) availability; therefore, the down-regulation of MK2 diminishes PIP 3 production and decreases Akt activity [19]. PI3K/Akt signaling plays an important role in cell survival [20, 21].…”

Section: Introductionmentioning

confidence: 99%

Insulin Influences LPS-Induced TNF-α and IL-6 Release Through Distinct Pathways in Mouse Macrophages from Different Compartments

Tessaro¹,

Ayala²,

Nolasco³

et al. 2017

Cell Physiol Biochem

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Background/Aims: Diabetic subjects are more susceptible to infections, which is partially due to insulin deficiency and hyperglycemia. We hypothesized that insulin influences cytokine release by macrophages from diabetic C57BL/6 mice stimulated with lipopolysaccharides (LPS). Methods: Bone marrow-derived macrophages (BMDM) and tissue-specific macrophages from diabetic (alloxan 60 mg/kg, i.v.) male C57BL/6 mice were stimulated by LPS (100 ng/mL) and/or treated by insulin (1 mU/mL). Results: Using BMDM from diabetic mice, we showed that LPS induced an increase in TNF-α and IL-6 release and p38, SAPK/JNK, ERK 1/2, and Akt (308-Thr and 473-Ser) phosphorylation but not in PKCα/β II and delta. Insulin increased TNF-α and IL-6 secretion in LPS-stimulated macrophages as well as p-p38, p-SAPK/JNK, p-ERK 1/2, p-PI3K (p55) and p-Akt (473-Ser) expression. Furthermore, PI3-kinase inhibition by wortmannin decreased TNF-α release, and inhibition by LY294002 decreased both TNF-α and IL-6 levels after LPS-insulin treatment. PD98059, which inhibits the ERK upstream activators MAPK kinase (MKK) 1 and MKK2, reduced the effect promoted by insulin in BMDM stimulated by LPS In tissue-specific macrophages, insulin reduced LPS-induced TNF-α, IL-6 and IL-1β secretion in alveolar and peritoneal macrophages. Conclusion: These data suggest that insulin through the modulation of PI3-kinase and ERK 1/2 pathways drive different responses in macrophages, thereby enhancing our understanding of the plasticity of these cells.

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“…6). The MAPK and AKT pathways are important downstream pathways of LPS/TLR signaling (31). To elucidate the regulatory mechanisms of YCG063 involved in these processes, we investigated whether the AKT and MAPK signaling pathways are involved in regulating the inflammatory mediators.…”

Section: Discussionmentioning

confidence: 99%

YCG063 inhibits Pseudomonas aeruginosa LPS-induced inflammation in human retinal pigment epithelial cells through the TLR2-mediated AKT/NF-κB pathway and ROS-independent pathways

Paeng

Park

Jung

et al. 2015

International Journal of Molecular Medicine

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Abstract. YCG063 is known as an inhibitor of reactive oxygen species (ROS); however, its intracellular mechanisms of action remain poorly understood. In the present study, we investigated the effects of YCG063 on the inflammatory response of Pseudomonas aeruginosa lipopolysaccharide (PA-LPS)-stimulated human retinal pigment epithelial cells (RPE cells). Human adult RPE cells (ARPE-19) were stimulated with PA-LPS. We then investigated the LPS-induced expression of several inflammatory mediators, such as interleukin (IL)-6, IL-8, monocyte chemoattractant protein-1 (MCP-1) and intracellular adhesion molecule-1 (ICAM-1) in the ARPE-19 cells. We performed an enzyme-linked immunosorbent assay (ELISA), western blot analysis, electrophoretic mobility shift assay (EMSA) and fluorescence-activated cell sorting (FACS) to elucidate the mechanisms involved in the anti-inflammatory effects of YCG063 in the PA-LPS-stimulated cells. The results revealed that treatment with YCG063 significantly inhibited the levels of IL-6, IL-8, MCP-1 and ICAM-1 in the PA-LPS-stimulated ARPE-19 cells. YCG063 also markedly inhibited the phosphorylation of AKT in the PA-LPS-stimulated cells. In addition, the activation of nuclear factor-κB (NF-κB) was also attenuated folllowing treatment with YCG063. ROS were not generated in the PA-LPS-stimulated cells. In conclusion, our data indicate that YCG063 may prove to be a potential protective agent against inflammation, possibly through the downregulation of Toll-like receptor 2 (TLR2) and the AKT-dependent NF-κB activation pathway in PA-LPSstimulated ARPE-19 cells. Furthermore, this anti-inflammatory activity occurred through ROS-independent signaling pathways. IntroductionEndophthalmitis is an uncommon intraocular inflammatory condition, but it may result in partial or complete vision loss. It may occur as a complication of intraocular surgery or as a result of non-surgical trauma or systemic infection (1). Bacterial endophthalmitis is an infection of the intraocular cavities that usually occurs following the introduction of microbial organisms into the eye (2). During a bacterial infection, irreversible damage to the photoreceptor cells of the retina frequently occurs (3). Among the bacterial pathogens, Pseudomonas aeruginosa (PA), a Gram-negative rod rod-shaped organism, is frequently a cause of nosocomial infections (4). PA can cause ocular infections, such as keratitis and endophthalmitis (5). YCG063 inhibits Pseudomonas aeruginosa LPS-induced inflammation in human retinal pigment epithelial cells through the TLR2-mediated AKT/NF-κB pathway and ROS-independent pathways

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Cross Talk between the Akt and p38α Pathways in Macrophages Downstream of Toll-Like Receptor Signaling

Cited by 67 publications

References 76 publications

Development of Selective Covalent Janus Kinase 3 Inhibitors

Development of Selective Covalent Janus Kinase 3 Inhibitors

Insulin Influences LPS-Induced TNF-α and IL-6 Release Through Distinct Pathways in Mouse Macrophages from Different Compartments

YCG063 inhibits Pseudomonas aeruginosa LPS-induced inflammation in human retinal pigment epithelial cells through the TLR2-mediated AKT/NF-κB pathway and ROS-independent pathways

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