bluuebunny/crossref_metadata_2025_split huggingface dataset:
Co‐activation of adenosine and glutamate receptors modulates signaling in primary brain cell cultures
Authors: Gregory Karen J. , Hellyer Shane D. , Albold Sabine , Leach Katie , May Lauren T. 
Published: June 2021

G protein‐coupled receptors (GPCRs) represent the largest class of targets for current therapeutics. Two distinct modulatory GPCRs within the CNS, the adenosine A1 receptor (A1AR) and the metabotropic glutamate receptor subtype 5 (mGlu5), have been implicated in Alzheimer's disease (AD) pathology and as potential targets to treat cognitive impairments and disease progression. Both GPCRs are present on the same CNS‐resident cell types, e.g. neurons and astrocytes, and are expressed in brain regions implicated in AD pathology. GPCR discovery programs generally only consider GPCR activity in isolation, without factoring in the influence of other GPCRs or stimuli present. Of note, both glutamate and adenosine are often present within culture medium and/or released from cultured cells. GPCR cross‐talk and/or heteromerization can introduce pharmacological heterogeneity and offer new avenues for targeted drug development. We sought to test the hypothesis that coincident activation of co‐located GPCRs modulates signaling in primary cultures of CNS resident cells. Primary cultures were derived from striatum and cortices of E16 mice and cultured for 6–8 days. High‐throughput signaling assays (iCa2+ mobilization, inositol phosphate (IP1) and cAMP accumulation) and radioligand binding studies were performed to confirm expression and function of mGlu5 and A1AR in the cultures by assessing the ability of: 1) A1AR selective (MeCCPA) and non‐selective (NECA) agonists to inhibit forskolin stimulation of cAMP accumulation; and 2) mGlu5 selective allosteric agonist (VU0424465) and group I mGlu selective orthosteric agonist (DHPG) to stimulate iCa2+ and IP1 accumulation. Experiments performed in the presence and absence of adenosine deaminase revealed that ambient adenosine was not influencing mGlu5 signalling. Coincident activation of A1AR enhanced mGlu5‐mediated iCa2+ mobilization in response to both orthosteric and allosteric agonists in primary striatal and cortical neuronal cultures. However, A1AR activation had no effect on mGlu5‐mediated IP1 accumulation in cortical neurons. Conversely, coincident activation of mGlu5 had little influence on A1AR‐mediated inhibition of cAMP accumulation, but inhibited AR‐stimulation of ERK1/2 phosphorylation. Collectively, our data demonstrate that coincident activation of mGlu5 and A1AR differentially modulates intracellular signaling pathways. Future work exploring the underlying mechanisms may reveal new strategies for targeting these GPCRs to treat AD and cognitive disorders.Support or Funding InformationThis work was supported by National Health and Medical Research Council of Australia (APP1123722 to KJG and LTM) and Australian Research Council Future Fellowships to KJG and KL.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Seizure‐protective Receptors GPR37L1 and GPR37 Regulate Calcium‐binding Protein S100A5 Expression and Secretion
Authors: Nguyen TrangKimberly Thu , Giddens Michelle , Duong Duc , Hall Randy 
Published: June 2021

ObjectiveThe objective of this study is to investigate the mechanisms through which S100A5 expression and secretion is regulated by the receptors, GPR37L1 and GPR37.MethodsProteomic data obtained via mass spectrometry of mouse brain lysates was performed through the Emory Proteomics Core and follow‐up studies were performed studying cultured cells.Summary of resultsA mutation in the G protein‐coupled receptor GPR37L1 was recently found in a large family to be associated with a progressive form of epilepsy in which seizures start around puberty and increase in severity with age, ultimately resulting in death in the late teens to early twenties. Studies from our lab have revealed that knockout mice lacking GPR37L1 (and/or the closely related receptor GPR37) exhibit greatly increased susceptibility to seizures. To shed light on the potential mechanistic underpinnings of this phenotype, we performed proteomic analyses of whole mouse brain tissue from wild‐type vs. GPR37L1/GPR37 double knockout mice to identify proteins regulated by the absence versus presence of these receptors. The most dramatically downregulated protein in the knockout versus wild‐type brain tissue was S100A5, a brain‐specific calcium‐binding protein. The reduction of S100A5 expression in the knockout brain tissue was confirmed via Western blot. Furthermore, when S100A5 is co‐expressed with either GPR37L1 or GPR37 in HEK293T cells, a robust increase in secretion of S100A5 was observed. This secretion was receptor‐dependent, as over‐expression of S100A5 alone did not result in detectable S100A5 in the media. To dissect the G protein signaling pathway that mediates this secretion, inhibitors of the different G protein signaling pathways were used to treat cells that co‐expressed S100A5 with either receptor. These studies suggested that GPR37 and GPR37L1 signal through Gs to mediate S100A5 secretion, as treatment with H89, a protein kinase A (PKA) inhibitor, sharply reduces S100A5 secretion from transfected HEK293T cells. These findings may be of general interest because other members of the S100A protein family are known to be secreted and have neuroprotective actions within the brain, but little is known about the mechanisms controlling their secretion.ConclusionsS100A5 expression and secretion are both regulated by GPR37 and GPR37L1. Our studies reveal that these receptors signal in a PKA‐dependent manner to regulate S100A5 secretion.Support or Funding InformationPharmacological Sciences T32 Predoctoral Training Grant (4T32GM008602‐20), Pharmacological Sciences T32 Predoctoral Training Grant (4T32GM008602‐21), NIH Grants R21‐NS91986 and R01‐NS088413This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
EET‐agonist Prevents and Reverses Heart Failure in Obesity Induced Diabetic Cardiomyopathy
Authors: Singh Shailendra P. , McClung John A. , Bellner Lars , Schragenheim Joseph , Abraham Nader G. 
Published: June 2021

Previously we demonstrated that an EET‐agonist has pleiotropic effects including inhibition of Soluble Epoxide Hydrolase expression and reversed cardiomyopathy by decreasing inflammatory molecules and increasing antioxidant signaling. We hypothesized that an EET agonist would increase PGC‐1α, which controls mitochondrial function and HO‐1 induction and negatively regulates the expression of the proinflammatory adipokine NOV in both cardiac and pericardial tissues. This pathway would be expected to further improve left ventricular systolic function as well as increase insulin receptor phosphorylation. The effect of an EET agonist was measured in cardiac and pericardial adipose tissues in db/db mice on oxygen consumption, fractional shortening, blood glucose levels, thermogenic and mitochondrial signaling proteins. Control db/db mice developed signs of metabolic syndrome including insulin resistance, hypertension, inflammation, LV dysfunction, and increased NOV expression in pericardial adipose tissue. EET agonist decreased pericardial adipose tissue expression of NOV, normalized FS, increased PGC‐1α, HO‐1 levels, insulin receptor phosphorylation and improved mitochondrial function. HO‐1‐ derived carbon monoxide (CO) levels, mitochondrial function (p<0.01), insulin receptor phosphorylation (p<0.05), ATP levels and VO2 (p<0.05). Deletion of PGC‐1α reversed all of the above beneficial effects, increasing the expression of NOV and inflammatory cytokines in pericardial adipose tissue. These studies demonstrate that an EET agonist increases insulin receptor phosphorylation, mitochondrial and thermogenic gene expression, decreases cardiac and pericardial tissue NOV levels, and ameliorates cardiomyopathy in an obese mouse model of the metabolic syndrome. Further, this study demonstrated that EET‐mediated inhibition of NOV is a potential therapeutic target for the prevention of heart failure and the development of metabolic syndrome.Support or Funding InformationNational Institutes of Health grant (HL34300 to NGA)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Podocyte‐Specific Deletion of Asah1 Gene Produced Podocytopathy without Sclerotic Pathology: A Potential Mouse Model for Steroid‐Insensitive Minimal Change Disease
Authors: Li Guangbi , Lohner Hannah , Dempsey Sara , Yuan Xinxu , Zhang Qinghua , Gehr Todd , Ritter Joseph , Li Pin‐Lan 
Published: June 2021

Lysosomal acid ceramidase (AC) has been shown to be critical for the ceramide hydrolysis and thereby regulates lysosome function and related cell function such as autophagy and vesicle/molecular trafficking. Despite that ceramide is involved in the regulation of glomerular function, it remains unknown whether AC activity contributes to the control of podocyte function and whether its deficiency causes glomerular disease. In the present study, we generated a mouse line with podocyte‐specific deletion of α subunit in Asah1 (AC gene code in mouse) gene, a main subunit for its activity in lysosomes using Cre‐Lox recombination technology. This Asah1fl/fl/PodoCre mouse colony was characterized by several genetic, molecular and biochemical approaches. AC α subunit was not detectable in podocytes in glomeruli of Asah1fl/fl/PodoCre mice, compared with their control littermates (Asah1fl/fl/WT and WT/WT mice). Using LC‐MS/MS, remarkable ceramide accumulation was detected in isolated glomeruli of Asah1fl/fl/PodoCre mice, but not in their control littermates (increased from 42.99 pg/glomerulus to 237.19 pg/glomerulus). Although Periodic acid‐Schiff staining showed no significant increase in glomerular damage index in Asah1fl/fl/PodoCre mice compared with other two types of mice, severe proteinuria and albuminuria were found in Asah1fl/fl/PodoCre mice started at 8 weeks old (increased from 28.84 μg/24h/gbw to 181.41 μg/24h/gbw of urinary protein excretion, increased from 1.22 μg/24h/gbw to 10.39 μg/24h/gbw of urinary albumin excretion). Correspondingly, glomerular permeability to albumin was markedly increased by podocyte‐specific deletion of AC α subunit. Moreover, hypoalbuminemia and edema were observed in Asah1fl/fl/PodoCre mice when they grew to 12 weeks old. Under transmission electron microscope, Asah1fl/fl/PodoCre mice were found to have foot process effacement, vacuolation, and microvillus formation in podocytes. Treatment with dexamethasone (DEX) for 4 weeks failed to reverse podocyte injury and proteinuria in Asah1fl/fl/PodoCre mice. Immunohistochemistry showed no changes in α‐dystroglycan, but decreases in β‐dystroglycan in glomeruli of Asah1fl/fl/PodoCre mice, compared with their control littermates. These results suggest that lysosomal AC and associated ceramide hydrolysis is essential for the maintenance of podocyte structural and functional integrity and that the defect of AC expression and its function deficiency may result in podocytopathy and related glomerular disease such as steroid‐resistant minimal change disease.Support or Funding InformationThis study was supported by NIH grants DK54927 to Pin‐Lan Li and DK102539 to Joseph K. Ritter.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Contribution of High Mobility Group Box 1 to Obesity‐Induced Podocyte Dysfunction and Glomerular Injury
Authors: Koka Saisudha , Mohammad Razia S. , Singh Gurinder Bir , Kshirasagar Naresh , Hussain Tahir , Li Ningjun , Li Pin‐Lan , Li Xiang , Boini Krishna M. 
Published: June 2021

High mobility group box 1 protein (HMGB1), a nuclear DNA binding protein is released under pathological conditions and locally act as one of potent damage‐associated molecular patterns (DAMPs) to produce tissue injury and chronic inflammation. However, it remains unknown whether HMGB1 is implicated in obesity‐induced podocyte injury. In the present study, we found that obesity‐induced adipokine visfatin dose‐dependently increased the production of HMGB1 in cultured podocytes compared to control cells. However, pretreatment with HMGB1 binder, glycyrrhizin (Gly) completely blocked HMGB1 release which was induced by visfatin. Further, immunofluorescence analysis showed that visfatin treatment significantly decreased the podocin expression (podocyte damage) compared to control cells. However, prior treatment with glycerrhizin attenuated the visfatin‐induced podocin reduction. Furthermore, glycyrrhizin treatment was found to preserve podocyte morphology by maintaining the distinct arrangement of F‐actin fibers normally lost in response to visfatin. It also prevented podocyte dysfunction by restoring visfatin‐induced suppression of VEGF production and secretion. In addition, functional studies showed that dextran flux was significantly increased in visfatin treated podocytes compared to control cells. This visfatin‐induced increase in permeability was markedly reduced in the presence of glycyrrhizin. In in vivo studies, C57BL6 mice were fed with high fat diet or normal chow for 14 weeks to produce obesity. Western blot analysis showed that high fat diet treatment significantly increased the HMGB1, TLR4 and RAGE expression in glomeruli of high fat diet fed mice compared to normal diet fed mice. However, such increase of HMGB1, TLR4 and RAGE expression were attenuated in glomeruli of glycyrrhizin treated mice. Based on these results, it is concluded that HMGB1 is one of important mediators of obesity‐induced podocyte injury and glomerular sclerosis. HMGB1 may be a therapeutic target for treatment or prevention of glomerulosclerosis associated with obesity.Support or Funding Informationsupported by NIH grant, DK104031This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Upregulation of cystathionine‐γ‐lyase/hydrogen sulfide pathway underlies the celecoxib counteraction of the cyclosporine‐induced hypertension and renal insult in rats
Authors: Helmy Mai M. , Helmy Maged Wasfy , El‐Mas Mahmoud M. 
Published: June 2021

We recently reported that celecoxib, a selective cyclooxygenase‐2 inhibitor, counteracts the adverse circulatory and renal actions of cyclosporine (CSA). Despite the advantageous nature of this interaction particularly when the two drugs are prescribed together in the management of immune‐related arthritic conditions, the underlying mechanism remains elusive. This prompted us to test the hypothesis that the facilitation of the cystathionine‐γ‐lyase (CES)/hydrogen sulfide (H2S) signaling accounts for the celecoxib‐CSA interaction. The data showed that the 10‐day cotreatment of rats with celecoxib (10 mg/kg/day) ameliorated the hypertensive and biochemical and structural renal damages caused by CSA (20 mg/kg/day). Celecoxib also reversed the CSAevoked (i) reductions in the tubular and glomerular protein expression of CES and levels of H2S, prostaglandin E2, and total antioxidant capacity, and (iii) increases in tumor necrosis factor‐α, transforming growth factor‐β1 and caspase‐3. These favorable effects of celecoxib disappeared when rats were treated concomitantly with the CES inhibitor DL‐propargylglycine (PAG), indicating the importance of the CES‐derived H2S in mediating the celecoxib counteraction of the CSA effects. This view is bolstered by the observation that the beneficial hemodynamic and renal actions of celecoxib were replicated after supplementation of rats with sodium sulfide (H2S donor). Together, the increased abundance of renal CES and H2S generation and subsequent dampening down of inflammatory, fibrotic, oxidant, and apoptotic pathways play pivotal roles in the celecoxib counteraction of the troublesome hypetensive and nephrotoxic insults caused by CSA in rats.Support or Funding Informationthe running research was funded by authors themselves.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Calcium Sulfide Nanoclusters in the Migration of Non‐Small Cell Lung Adenocarcinoma cell line HCC827
Authors: Reyes Amalia Marie Rivera , Forti Kevin Muñoz , Martínez Gerardo Arroyo , Torres Geraline Trossi , Rivera Abigail Arroyo Ruiz , Suarez‐Martínez Edu Arroyo 
Published: June 2021

Lung cancer is the most common malignancy worldwide independently of gender. Non‐small cell lung carcinoma (NSCLC) is responsible for 80% of diagnosed cases. Prognosis for patients suffering this cancer is poor, with only 7% of patients surviving five years after the diagnosis. Current treatments include surgery, radiotherapy, and chemotherapy. Although the aforementioned seem successful, they do not address the development of secondary tumors due to cell migration resulting in metastasis. Evidently, there is a need for developing novel therapeutic approaches specifically targeting malignant cells. In our laboratory, we evaluate calcium sulfide (CaS) nanostructures as potential multi‐cancer malignant‐cell targeted specific treatment. The objective of this study was to determine the effect of CaS nanoclusters in the migration of NSCLC HCC827 cells. We hypothesized that CaS will decrease the area of migration compared to the control vehicle (DMSO), will increase the number of dead cells and decrease the live cells in the collected supernatant. We used the Scratch Wound Healing Assay, to determine the area of migration after 24, 48, and 72hrs upon a single exposure at time 0 of CaS treatment and controls (DMSO: vehicle control; etoposide: positive control; and media: negative control). We also collected the supernatant to characterize detached cells in suspension. Preliminary data shows that at 72hrs, cells treated with CaS exhibited a significant decrease in cell migration. We categorized the cells in the supernatant as live or dead. Although there were not statistically significant differences when compared CaS treatment with the controls, we observed a time‐dependent trend. This trend showed a decrease of the number of dead cells and an increase of live cells as the time increases. Based on this preliminary data, we partly accept our hypothesis due to the observed decreasing of migration in CaS treatment. However, the supernatant data was the opposite to what we hypothesized. We are currently optimizing experiments to increase statistical power and correlate the area of migration with the supernatant findings. In the next step of this project, I will be determining the effect of CaS by means of migration and number of live and dead cells in the supernatant in the non‐malignant lung cells MRC5.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
G Protein‐Coupled Receptors in Human Lung Fibroblasts: Novel Therapeutic Approaches for Idiopathic Pulmonary Fibrosis?
Authors: Michkov Alexander V. , Sriram Krishna , Mukherjee Subhendu , Janssen Luke J. , Insel Paul A. 
Published: June 2021

Idiopathic Pulmonary Fibrosis (IPF) is a progressive, often fatal interstitial lung disease and thus in need of new, effective therapies. A feature of IPF is fibroblast‐to‐myofibroblast transition and resultant increased deposition of extracellular matrix (ECM) proteins, which can lead to loss of functional lung tissue. Previous studies have shown that increases in cyclic AMP (cAMP) can decrease the number and function of myofibroblasts. Cellular synthesis of cAMP is mediated by adenylyl cyclase, the activity of which is regulated by G‐protein signaling and G‐protein coupled receptors (GPCRs). We hypothesized that alteration in GPCR expression in lung fibroblasts from IPF patients may identify new therapeutic targets for lung fibrosis. Using RNA‐seq, we have identified and quantified GPCRs of lung fibroblasts isolated from six IPF patients and six controls grown in low passage primary cultures. RNA‐seq analysis revealed the expression of ~120 GPCRs (of ~380 endoGPCRs [regulated by endogenous ligands]). On average ~20 highly expressed GPCR were detected in all samples, with small differences in biological replicates; these may be “housekeeping” GPCRs. We also identified a group of GPCRs with highly variable expression and found that lung fibroblasts from IPF patients selectively expressed a subset of GPCRs. To assess changes in GPCR expression during fibroblast‐to‐myofibroblast transition, we treated fibroblasts with TGFβ1 (10 ng/ml, 48 hrs or 20 ng/ml, 16 hrs). As expected, TGFβ1 treatment increased the expression of pro‐fibrotic markers (e.g., collagens Iα1, Iα2, III; α‐smooth muscle actin; plasminogen activator inhibitor‐1). In addition, TGFβ1 altered (increased or decreased) the expression of numerous GPCRs. TGFβ1 treatment of control fibroblasts did not fully mimic the GPCR expression changes of fibroblasts from IPF patients. We conclude that GPCR expression (GPCRomic) analysis identifies GPCRs that may contribute to the fibrotic state in the lung and thus, may be novel therapeutic targets for lung fibrosis.Support or Funding InformationSupported by NIH T32 HL 098062‐04, Bristol Myers Squibb and CIHR 201403MOP‐RS‐323925.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Resolvin D1 blocks ROS‐mediated inhibitory crosstalk between SHP2 and PP2A and suppresses endothelial‐monocyte interactions
Authors: Singh Nikhlesh , Chattopadhyay Rima N. , Mani Arul M. , Rao Gadiparthi N. 
Published: June 2021

In recent years, various studies have demonstrated a role for endogenously derived specialized lipid mediators such as resolvins in the resolution of inflammation. In exploring the signaling mechanisms, in the present study, we show that RvD1 reduces LPS‐induced endothelial cell (EC)‐monocyte interactions via blocking ROS‐mediated PP2A inactivation, NFκB activation and ICAM1 and VCAM1 expression. In addition, we found that ROS‐mediated SHP2 inhibition leads to tyrosine phosphorylation and inactivation of PP2A by LPS, which in turn, accounts for increased NFκB activation and ICAM1 and VCAM1 expression facilitating EC‐monocyte interactions and all these LPS‐mediated responses were reversed by RvD1. Furthermore, the suppression of NFκB activation, ICAM1 and VCAM1 expression and EC and monocyte interactions by RvD1 involved its receptors ALX/FPR2 and GPR32 as inhibition or neutralization of these receptors negated its effects. Together, these observations show that RvD1 via protecting SHP2 from ROS‐mediated inactivation blocks PP2A inactivation and thereby shuts of NFκB activation, ICAM1 and VCAM1 expression and EC‐monocyte interactions, which could be one of the several possible mechanisms underlying its anti‐inflammatory functions.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.