101 results on '"Olivier Mignen"'
Search Results
2. LKB1‐SIK2 loss drives uveal melanoma proliferation and hypersensitivity to SLC8A1 and ROS inhibition
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Sarah Proteau, Imène Krossa, Chrystel Husser, Maxime Guéguinou, Federica Sella, Karine Bille, Marie Irondelle, Mélanie Dalmasso, Thibault Barouillet, Yann Cheli, Céline Pisibon, Nicole Arrighi, Sacha Nahon‐Estève, Arnaud Martel, Lauris Gastaud, Sandra Lassalle, Olivier Mignen, Patrick Brest, Nathalie M Mazure, Frédéric Bost, Stéphanie Baillif, Solange Landreville, Simon Turcotte, Dan Hasson, Saul Carcamo, Christophe Vandier, Emily Bernstein, Laurent Yvan‐Charvet, Mitchell P Levesque, Robert Ballotti, Corine Bertolotto, and Thomas Strub
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calcium ,LKB1 ,SIK2 ,SLC8A1 ,uveal melanoma ,Medicine (General) ,R5-920 ,Genetics ,QH426-470 - Abstract
Abstract Metastatic uveal melanomas are highly resistant to all existing treatments. To address this critical issue, we performed a kinome‐wide CRISPR‐Cas9 knockout screen, which revealed the LKB1‐SIK2 module in restraining uveal melanoma tumorigenesis. Functionally, LKB1 loss enhances proliferation and survival through SIK2 inhibition and upregulation of the sodium/calcium (Na+/Ca2+) exchanger SLC8A1. This signaling cascade promotes increased levels of intracellular calcium and mitochondrial reactive oxygen species, two hallmarks of cancer. We further demonstrate that combination of an SLC8A1 inhibitor and a mitochondria‐targeted antioxidant promotes enhanced cell death efficacy in LKB1‐ and SIK2‐negative uveal melanoma cells compared to control cells. Our study also identified an LKB1‐loss gene signature for the survival prognostic of patients with uveal melanoma that may be also predictive of response to the therapy combination. Our data thus identify not only metabolic vulnerabilities but also new prognostic markers, thereby providing a therapeutic strategy for particular subtypes of metastatic uveal melanoma.
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- 2023
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3. Design and Synthesis of Novel N-Benzylidene Derivatives of 3-Amino-4-imino-3,5-dihydro-4H-chromeno[2,3-d]pyrimidine under Microwave, In Silico ADME Predictions, In Vitro Antitumoral Activities and In Vivo Toxicity
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Sirine Karoui, Marwa Dhiabi, Mehdi Fakhfakh, Souhir Abid, Emmanuelle Limanton, Rémy Le Guével, Thierry D. Charlier, Anthony Mainguy, Olivier Mignen, Ludovic Paquin, Houcine Ammar, and Jean-Pierre Bazureau
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2-amino-4H-chromene ,4H-chromeno[2,3-d]pyrimidine ,microwave irradiation ,tumoral cell line ,cytotoxicity ,Lipinski’s rules of five ,Medicine ,Pharmacy and materia medica ,RS1-441 - Abstract
The synthesis of a series of new N-benzylidene derivatives of 3-amino-4-imino-3,5-dihydro-4H-chromeno[2,3-d]pyrimidine 10(a-l) bearing two points of molecular diversity is reported. These new compounds were synthesized in five steps including two steps under microwave dielectric heating. They were fully characterized using 1H and 13C NMR, FTIR and HRMS. The in silico physicochemical properties of compounds 10(a-l) were determined according to Lipinski’s rules of five (RO5) associated with the prediction of their bioavailability. These new compounds 10(a-l) were tested for their antiproliferative activities in fibroblasts and eight representative human tumoral cell lines (Huh7 D12, Caco2, MDA-MB231, MDA-MB468, HCT116, PC3, MCF7 and PANC1). Among them, the compounds 10h and 10i showed sub-micromolar cytotoxic activity on tumor cell lines (0.23 < IC50 < 0.3 μM) and no toxicity on fibroblasts (IC50 > 25 μM). A dose-dependent inhibition of Store-Operated Ca+2 Entry (SOCE) was observed in the HEK293 cell line with 10h. In vitro embryotoxicity and angiogenesis on the mCherry transgenic zebrafish line showed that 10h presented no toxic effect and no angiogenic effect on embryos with a dose of 5 μM at 72 hpf.
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- 2024
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4. New insights into the regulation of Cystathionine beta synthase (CBS), an enzyme involved in intellectual deficiency in Down syndrome
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Pierre Conan, Alice Léon, Noéline Caroff, Claire Rollet, Loubna Chaïr, Jennifer Martin, Frédéric Bihel, Olivier Mignen, Cécile Voisset, and Gaëlle Friocourt
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CBS ,DYRK1A ,GSK3β ,Akt ,NF-κB ,pharmacological inhibitor ,Neurosciences. Biological psychiatry. Neuropsychiatry ,RC321-571 - Abstract
Down syndrome (DS), the most frequent chromosomic aberration, results from the presence of an extra copy of chromosome 21. The identification of genes which overexpression contributes to intellectual disability (ID) in DS is important to understand the pathophysiological mechanisms involved and develop new pharmacological therapies. In particular, gene dosage of Dual specificity tyrosine phosphorylation Regulated Kinase 1A (DYRK1A) and of Cystathionine beta synthase (CBS) are crucial for cognitive function. As these two enzymes have lately been the main targets for therapeutic research on ID, we sought to decipher the genetic relationship between them. We also used a combination of genetic and drug screenings using a cellular model overexpressing CYS4, the homolog of CBS in Saccharomyces cerevisiae, to get further insights into the molecular mechanisms involved in the regulation of CBS activity. We showed that overexpression of YAK1, the homolog of DYRK1A in yeast, increased CYS4 activity whereas GSK3β was identified as a genetic suppressor of CBS. In addition, analysis of the signaling pathways targeted by the drugs identified through the yeast-based pharmacological screening, and confirmed using human HepG2 cells, emphasized the importance of Akt/GSK3β and NF-κB pathways into the regulation of CBS activity and expression. Taken together, these data provide further understanding into the regulation of CBS and in particular into the genetic relationship between DYRK1A and CBS through the Akt/GSK3β and NF-κB pathways, which should help develop more effective therapies to reduce cognitive deficits in people with DS.
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- 2023
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5. Molecular Mechanisms Driving IL-10- Producing B Cells Functions: STAT3 and c-MAF as Underestimated Central Key Regulators?
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Magalie Michée-Cospolite, Marina Boudigou, Alexis Grasseau, Quentin Simon, Olivier Mignen, Jacques-Olivier Pers, Divi Cornec, Laëtitia Le Pottier, and Sophie Hillion
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regulatory B cells ,molecular drivers ,c-MAF ,STAT3 ,B-cell differentiation ,Immunologic diseases. Allergy ,RC581-607 - Abstract
Regulatory B cells (Bregs) have been highlighted in very different pathology settings including autoimmune diseases, allergy, graft rejection, and cancer. Improving tools for the characterization of Bregs has become the main objective especially in humans. Transitional, mature B cells and plasma cells can differentiate into IL-10 producing Bregs in both mice and humans, suggesting that Bregs are not derived from unique precursors but may arise from different competent progenitors at unrestricted development stages. Moreover, in addition to IL-10 production, regulatory B cells used a broad range of suppressing mechanisms to modulate the immune response. Although Bregs have been consistently described in the literature, only a few reports described the molecular aspects that control the acquisition of the regulatory function. In this manuscript, we detailed the latest reports describing the control of IL-10, TGFβ, and GZMB production in different Breg subsets at the molecular level. We focused on the understanding of the role of the transcription factors STAT3 and c-MAF in controlling IL-10 production in murine and human B cells and how these factors may represent an important crossroad of several key drivers of the Breg response. Finally, we provided original data supporting the evidence that MAF is expressed in human IL-10- producing plasmablast and could be induced in vitro following different stimulation cocktails. At steady state, we reported that MAF is expressed in specific human B-cell tonsillar subsets including the IgD+ CD27+ unswitched population, germinal center cells and plasmablast.
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- 2022
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6. STIM1 at the plasma membrane as a new target in progressive chronic lymphocytic leukemia
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Marjolaine Debant, Miguel Burgos, Patrice Hemon, Paul Buscaglia, Tinhinane Fali, Sarra Melayah, Nelig Le Goux, Christophe Vandier, Marie Potier-Cartereau, Jacques-Olivier Pers, Adrian Tempescul, Christian Berthou, Cristina Bagacean, Olivier Mignen, and Yves Renaudineau
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CLL ,STIM1 ,Constitutive Ca2+ entry ,Disease outcome ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Abstract Background Dysregulation in calcium (Ca2+) signaling is a hallmark of chronic lymphocytic leukemia (CLL). While the role of the B cell receptor (BCR) Ca2+ pathway has been associated with disease progression, the importance of the newly described constitutive Ca2+ entry (CE) pathway is less clear. In addition, we hypothesized that these differences reflect modifications of the CE pathway and Ca2+ actors such as Orai1, transient receptor potential canonical (TRPC) 1, and stromal interaction molecule 1 (STIM1), the latter being the focus of this study. Methods An extensive analysis of the Ca2+ entry (CE) pathway in CLL B cells was performed including constitutive Ca2+ entry, basal Ca2+ levels, and store operated Ca2+ entry (SOCE) activated following B cell receptor engagement or using Thapsigargin. The molecular characterization of the calcium channels Orai1 and TRPC1 and to their partner STIM1 was performed by flow cytometry and/or Western blotting. Specific siRNAs for Orai1, TRPC1 and STIM1 plus the Orai1 channel blocker Synta66 were used. CLL B cell viability was tested in the presence of an anti-STIM1 monoclonal antibody (mAb, clone GOK) coupled or not with an anti-CD20 mAb, rituximab. The Cox regression model was used to determine the optimal threshold and to stratify patients. Results Seeking to explore the CE pathway, we found in untreated CLL patients that an abnormal CE pathway was (i) highly associated with the disease outcome; (ii) positively correlated with basal Ca2+ concentrations; (iii) independent from the BCR-PLCγ2-InsP3R (SOCE) Ca2+ signaling pathway; (iv) supported by Orai1 and TRPC1 channels; (v) regulated by the pool of STIM1 located in the plasma membrane (STIM1PM); and (vi) blocked when using a mAb targeting STIM1PM. Next, we further established an association between an elevated expression of STIM1PM and clinical outcome. In addition, combining an anti-STIM1 mAb with rituximab significantly reduced in vitro CLL B cell viability within the high STIM1PM CLL subgroup. Conclusions These data establish the critical role of a newly discovered BCR independent Ca2+ entry in CLL evolution, provide new insights into CLL pathophysiology, and support innovative therapeutic perspectives such as targeting STIM1 located at the plasma membrane.
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- 2019
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7. Identification of 8-Hydroxyquinoline Derivatives That Decrease Cystathionine Beta Synthase (CBS) Activity
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Pierre Conan, Alice Léon, Mathilde Gourdel, Claire Rollet, Loubna Chaïr, Noéline Caroff, Nelig Le Goux, Catherine Le Jossic-Corcos, Maha Sinane, Lucile Gentile, Louise Maillebouis, Nadège Loaëc, Jennifer Martin, Marie Vilaire, Laurent Corcos, Olivier Mignen, Mikael Croyal, Cécile Voisset, Frédéric Bihel, and Gaëlle Friocourt
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CBS ,drug screening ,Cys4 ,copper ,zinc ,cytosolic pH ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
CBS encodes a pyridoxal 5′-phosphate-dependent enzyme that catalyses the condensation of homocysteine and serine to form cystathionine. Due to its implication in some cancers and in the cognitive pathophysiology of Down syndrome, the identification of pharmacological inhibitors of this enzyme is urgently required. However, thus far, attempts to identify such molecules have only led to the identification of compounds with low potency and limited selectivity. We consequently developed an original, yeast-based screening method that identified three FDA-approved drugs of the 8-hydroxyquinoline family: clioquinol, chloroxine and nitroxoline. These molecules reduce CBS enzymatic activity in different cellular models, proving that the molecular mechanisms involved in yeast phenotypic rescue are conserved in mammalian cells. A combination of genetic and chemical biology approaches also revealed the importance of copper and zinc intracellular levels in the regulation of CBS enzymatic activity—copper promoting CBS activity and zinc inhibiting its activity. Taken together, these results indicate that our effective screening approach identified three new potent CBS inhibitors and provides new findings for the regulation of CBS activity, which is crucial to develop new therapies for CBS-related human disorders.
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- 2022
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8. Pacific-Ciguatoxin-2 and Brevetoxin-1 Induce the Sensitization of Sensory Receptors Mediating Pain and Pruritus in Sensory Neurons
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Ophélie Pierre, Maxime Fouchard, Nelig Le Goux, Paul Buscaglia, Raphaël Leschiera, Richard J. Lewis, Olivier Mignen, Joachim W. Fluhr, Laurent Misery, and Raphaële Le Garrec
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ciguatoxin ,ciguatera fish poisoning ,brevetoxin ,neurotoxic shellfish poisoning ,sensory disorders ,pruritus ,Biology (General) ,QH301-705.5 - Abstract
Ciguatera fish poisoning (CFP) and neurotoxic shellfish poisoning syndromes are induced by the consumption of seafood contaminated by ciguatoxins and brevetoxins. Both toxins cause sensory symptoms such as paresthesia, cold dysesthesia and painful disorders. An intense pruritus, which may become chronic, occurs also in CFP. No curative treatment is available and the pathophysiology is not fully elucidated. Here we conducted single-cell calcium video-imaging experiments in sensory neurons from newborn rats to study in vitro the ability of Pacific-ciguatoxin-2 (P-CTX-2) and brevetoxin-1 (PbTx-1) to sensitize receptors and ion channels, (i.e., to increase the percentage of responding cells and/or the response amplitude to their pharmacological agonists). In addition, we studied the neurotrophin release in sensory neurons co-cultured with keratinocytes after exposure to P-CTX-2. Our results show that P-CTX-2 induced the sensitization of TRPA1, TRPV4, PAR2, MrgprC, MrgprA and TTX-r NaV channels in sensory neurons. P-CTX-2 increased the release of nerve growth factor and brain-derived neurotrophic factor in the co-culture supernatant, suggesting that those neurotrophins could contribute to the sensitization of the aforementioned receptors and channels. Our results suggest the potential role of sensitization of sensory receptors/ion channels in the induction or persistence of sensory disturbances in CFP syndrome.
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- 2021
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9. Calcium Increase and Substance P Release Induced by the Neurotoxin Brevetoxin-1 in Sensory Neurons: Involvement of PAR2 Activation through Both Cathepsin S and Canonical Signaling
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Ophélie Pierre, Maxime Fouchard, Paul Buscaglia, Nelig Le Goux, Raphaël Leschiera, Olivier Mignen, Joachim W. Fluhr, Laurent Misery, and Raphaële Le Garrec
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brevetoxin ,ciguatoxin ,sensory disorders ,substance P ,cathepsin S ,PAR2 ,Cytology ,QH573-671 - Abstract
Red tides involving Karenia brevis expose humans to brevetoxins (PbTxs). Oral exposition triggers neurotoxic shellfish poisoning, whereas inhalation induces a respiratory syndrome and sensory disturbances. No curative treatment is available and the pathophysiology is not fully elucidated. Protease-activated receptor 2 (PAR2), cathepsin S (Cat-S) and substance P (SP) release are crucial mediators of the sensory effects of ciguatoxins (CTXs) which are PbTx analogs. This work explored the role of PAR2 and Cat-S in PbTx-1-induced sensory effects and deciphered the signaling pathway involved. We performed calcium imaging, PAR2 immunolocalization and SP release experiments in monocultured sensory neurons or co-cultured with keratinocytes treated with PbTx-1 or P-CTX-2. We demonstrated that PbTx-1-induced calcium increase and SP release involved Cat-S, PAR2 and transient receptor potential vanilloid 4 (TRPV4). The PbTx-1-induced signaling pathway included protein kinase A (PKA) and TRPV4, which are compatible with the PAR2 biased signaling induced by Cat-S. Internalization of PAR2 and protein kinase C (PKC), inositol triphosphate receptor and TRPV4 activation evoked by PbTx-1 are compatible with the PAR2 canonical signaling. Our results suggest that PbTx-1-induced sensory disturbances involve the PAR2-TRPV4 pathway. We identified PAR2, Cat-S, PKA, and PKC that are involved in TRPV4 sensitization induced by PbTx-1 in sensory neurons.
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- 2020
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10. Neurological Disturbances of Ciguatera Poisoning: Clinical Features and Pathophysiological Basis
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Killian L’Herondelle, Matthieu Talagas, Olivier Mignen, Laurent Misery, and Raphaele Le Garrec
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ciguatera ,ciguatoxin ,neurological ,sensory ,pathophysiology ,Cytology ,QH573-671 - Abstract
Ciguatera fish poisoning (CFP), the most prevalent seafood poisoning worldwide, is caused by the consumption of tropical and subtropical fish contaminated with potent neurotoxins called ciguatoxins (CTXs). Ciguatera is a complex clinical syndrome in which peripheral neurological signs predominate in the acute phase of the intoxication but also persist or reoccur long afterward. Their recognition is of particular importance in establishing the diagnosis, which is clinically-based and can be a challenge for physicians unfamiliar with CFP. To date, no specific treatment exists. Physiopathologically, the primary targets of CTXs are well identified, as are the secondary events that may contribute to CFP symptomatology. This review describes the clinical features, focusing on the sensory disturbances, and then reports on the neuronal targets and effects of CTXs, as well as the neurophysiological and histological studies that have contributed to existing knowledge of CFP neuropathophysiology at the molecular, neurocellular and nerve levels.
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- 2020
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11. In Vitro Differentiation of Human Skin-Derived Cells into Functional Sensory Neurons-Like
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Adeline Bataille, Raphael Leschiera, Killian L’Hérondelle, Jean-Pierre Pennec, Nelig Le Goux, Olivier Mignen, Mehdi Sakka, Emmanuelle Plée-Gautier, Cecilia Brun, Thierry Oddos, Jean-Luc Carré, Laurent Misery, and Nicolas Lebonvallet
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sensory neuron ,adult stem cell ,differentiation ,TRP ,Cytology ,QH573-671 - Abstract
Skin-derived precursor cells (SKPs) are neural crest stem cells that persist in certain adult tissues, particularly in the skin. They can generate a large type of cell in vitro, including neurons. SKPs were induced to differentiate into sensory neurons (SNs) by molecules that were previously shown to be important for the generation of SNs: purmorphamine, CHIR99021, BMP4, GDNF, BDNF, and NGF. We showed that the differentiation of SKPs induced the upregulation of neurogenins. At the end of the differentiation protocol, transcriptional analysis was performed on BRN3A and a marker of pain-sensing nerve cell PRDM12 genes: 1000 times higher for PRDM12 and 2500 times higher for BRN3A in differentiated cells than they were in undifferentiated SKPs. Using immunostaining, we showed that 65% and 80% of cells expressed peripheral neuron markers BRN3A and PERIPHERIN, respectively. Furthermore, differentiated cells expressed TRPV1, PAR2, TRPA1, substance P, CGRP, HR1. Using calcium imaging, we observed that a proportion of cells responded to histamine, SLIGKV (a specific agonist of PAR2), polygodial (a specific agonist of TRPA1), and capsaicin (a specific agonist of TRPV1). In conclusion, SKPs are able to differentiate directly into functional SNs. These differentiated cells will be very useful for further in vitro studies.
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- 2020
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12. A Convenient Four-Step Synthesis of 1-{β-[3-(4-Methoxy-phenyl)Propoxy]-4-Methoxyphenethyl}-1H-Imidazole Hydrochloride as a Probing Tool for SOCE Assays
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Camille D. Dago, Estelle Messé, Olivier Mignen, Christophe Brigaudeau, Yves-Alain Békro, and Jean-Pierre Bazureau
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SKF-96365 ,imidazole ,modulator ,SOCE ,Inorganic chemistry ,QD146-197 - Abstract
1-{β-[3-(4-Methoxy-phenyl)propoxy]-4-methoxyphenethyl}-1H-imidazole hydrochloride 7, or SKF-96365, was synthesized in four steps with an overall yield of 9%. The structure of 7 was confirmed by 1H-, 13C-NMR, HRMS, and elemental analysis. The intermediates 3 and 4 were also isolated and characterized by 1H-, 13C-NMR, and HRMS.
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- 2016
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13. Preliminary Structure-Activity Relationship (SAR) of a Novel Series of Pyrazole SKF-96365 Analogues as Potential Store-Operated Calcium Entry (SOCE) Inhibitors
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Camille D. Dago, Paul Le Maux, Thierry Roisnel, Christophe Brigaudeau, Yves-Alain Bekro, Olivier Mignen, and Jean-Pierre Bazureau
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pyrazole ,SKF-96365 ,analogues ,resolution ,half-quantities ,SOCE ,B lymphocyte cell ,SOCE inhibitor ,Ca2+ signalling ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
From a series of (1R, 1S)-1[β-(phenylalkoxy)-(phenetyl)]-1H-pyrazolium hydrochloride as new analogues of SKF-96365, one has an interesting effect for endoplasmic reticulum (ER) Ca2+ release and store-operated Ca2+ entry (SOCE) (IC50 25 μM) on the PLP-B lymphocyte cell line. A successful resolution of (±) 1-phenyl-2-(1H-pyrazol-1-yl)ethan-1-ol has been developed by using the method of “half-concentration” in the presence of (+)-(1S)- or (−)-(1R)-CSA.
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- 2018
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14. Proteomic identification of calumenin as a G551D-CFTR associated protein.
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Ling Teng, Mathieu Kerbiriou, Mehdi Taiya, Sophie Le Hir, Olivier Mignen, Nathalie Benz, Pascal Trouvé, and Claude Férec
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Medicine ,Science - Abstract
Cystic fibrosis (CF) is the most common lethal autosomal recessive disease in the Caucasian population. It is due to mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. To date, over 1910 mutations have been identified in the CFTR gene. Among these mutations, the CF-causing missense mutation G551D-CFTR (approx. 5% of cases) encodes for a CFTR chloride channel with normal expression on the cell surface. Nevertheless, it is associated with severe disease due to its altered channel activation. The aim of the present study was to identify specific interacting proteins of G551D-CFTR. Co-immunoprecipitated proteins with G551D-CFTR were resolved by 2D-gel electrophoresis (2-DE). Mass Spectrometry revealed that calumenin was present in the protein complex linked to G551D-CFTR. Despite its basal expression was not modified in G551D-CFTR expressing cells when compared to Wt-CFTR expressing cells, it was more abundant in the G551D-CFTR complex detected by immunoprecipitation. The calumenin-CFTR interaction was also shown by Surface Plasmon Resonance and further confirmed by computational analysis of the predicted calumenin's partners. Because in our cellular model calumenin was found in the endoplasmic reticulum (ER) by immunofluorescence experiments, we suggest that calumenin is likely involved in the mutated CFTR's maturation. In conclusion, we showed for the first time that calumenin binds to CFTR and that it is increased in the G551D-CFTR complex. We suggest that it may be involved in the physiopathology of G551D-CFTR and that G551D-CFTR may follow a specific maturation and trafficking pathway. We also hypothesize that UPR may be triggered independently of the retention of G551D-CFTR in the ER because Grp78/Bip expression is increased in the cells. Finally, we propose here that Calumenin is a new CFTR chaperone.
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- 2012
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15. High-throughput analysis of promoter occupancy reveals new targets for Arx, a gene mutated in mental retardation and interneuronopathies.
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Marie-Lise Quillé, Solenne Carat, Sylvia Quéméner-Redon, Edouard Hirchaud, Daniel Baron, Caroline Benech, Jeanne Guihot, Morgane Placet, Olivier Mignen, Claude Férec, Rémi Houlgatte, and Gaëlle Friocourt
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Medicine ,Science - Abstract
Genetic investigations of X-linked intellectual disabilities have implicated the ARX (Aristaless-related homeobox) gene in a wide spectrum of disorders extending from phenotypes characterised by severe neuronal migration defects such as lissencephaly, to mild or moderate forms of mental retardation without apparent brain abnormalities but with associated features of dystonia and epilepsy. Analysis of Arx spatio-temporal localisation profile in mouse revealed expression in telencephalic structures, mainly restricted to populations of GABAergic neurons at all stages of development. Furthermore, studies of the effects of ARX loss of function in humans and animal models revealed varying defects, suggesting multiple roles of this gene during brain development. However, to date, little is known about how ARX functions as a transcription factor and the nature of its targets. To better understand its role, we combined chromatin immunoprecipitation and mRNA expression with microarray analysis and identified a total of 1006 gene promoters bound by Arx in transfected neuroblastoma (N2a) cells and in mouse embryonic brain. Approximately 24% of Arx-bound genes were found to show expression changes following Arx overexpression or knock-down. Several of the Arx target genes we identified are known to be important for a variety of functions in brain development and some of them suggest new functions for Arx. Overall, these results identified multiple new candidate targets for Arx and should help to better understand the pathophysiological mechanisms of intellectual disability and epilepsy associated with ARX mutations.
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- 2011
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16. Molecular Dynamics Simulations of Membrane-Bound STIM1 to Investigate Conformational Changes during STIM1 Activation upon Calcium Release.
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Sreya Mukherjee, Aleksandra Karolak, Marjolaine Debant, Paul Buscaglia, Yves Renaudineau, Olivier Mignen, Wayne C. Guida, and Wesley H. Brooks
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- 2017
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17. Active U11 Peptide Luminescent Gold Nanoclusters for Pancreatic Tumor Cell Targeting
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Regina M. Chiechio, Rémi Le Guevél, Solène Ducarre, Hélène Solhi, Stéphanie Dutertre, Xavier Pinson, Jean-Pierre Bazureau, Olivier Mignen, Pascale Even-Hernandez, Paolo Musumeci, Célia Ravel, Maria Josè Lo Faro, Valérie Marchi, Institut des Sciences Chimiques de Rennes (ISCR), Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Plate-forme ImPACcell (ImPACcell), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Institut de recherche en santé, environnement et travail (Irset), Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Biosit : biologie, santé, innovation technologique (SFR UMS CNRS 3480 - INSERM 018), Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Lymphocytes B, Autoimmunité et Immunothérapies (LBAI), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-LabEX IGO Immunothérapie Grand Ouest, Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO), and Università degli studi di Catania = University of Catania (Unict)
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U 11 peptide ,[SDV.IB.IMA]Life Sciences [q-bio]/Bioengineering/Imaging ,nucleus ,PANC cells ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,zebrafish ,peptide ,internalization ,one-pot synthesis ,[CHIM]Chemical Sciences ,General Materials Science ,gold nanoclusters ,targeting ,labeling - Abstract
International audience; The imaging of different intercellular regions is attracting growing interest in the fields of biosensing, drug delivery, and gene therapy for cancer treatment. We present the synthesis of luminescent gold nanoclusters (AuNCs), grafted with a PEGylated U 11 peptide derivative, for tumor recognition and their capacity to label the nucleus of pancreatic cancer cells. The short peptide named C 3 E 6 U 11 , used to synthesize and functionalize gold nanoclusters, is composed of a tricysteine sequence with sidechain thiol residues as peptide anchors for the Au nanoclusters together with the recognition peptide domain (U 11). The active targeting red-or blue-emitting AuNCs can label pancreatic cancer cells and localize preferentially in the nucleus. The presence of U 11 at their surface facilitates their penetration into pancreatic cell nuclei. Finally, in vivo imaging experiments in early embryos and freeswimming juvenile zebrafish are also performed to evaluate their biodistribution.
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- 2023
18. The diversity of the plasmablast signature across species and experimental conditions: A meta‐analysis
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Olivier Mignen, Sophie Hillion, Marina Boudigou, Magalie Michée-Cospolite, Laëtitia Le Pottier, Jacques-Olivier Pers, Christophe Jamin, Alexis Grasseau, Divi Cornec, Céline Delaloy, Lymphocyte B et Auto-immunité (LBAI), Université de Brest (UBO)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM), Microenvironment, Cell Differentiation, Immunology and Cancer (MICMAC), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Lymphocytes B, Autoimmunité et Immunothérapies (LBAI), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-LabEX IGO Immunothérapie Grand Ouest, Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO), Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), and ANR-18-CE15-0002,CascadingPCdiff,Signalisations B et engagement précoce vers la différenciation plasmocytaire(2018)
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Genetically modified mouse ,B-cell differentiation ,[SDV]Life Sciences [q-bio] ,Plasma Cells ,Immunology ,Mice, Transgenic ,Computational biology ,Biology ,Plasma cell ,Transcriptome ,Mice ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Humans ,Immunology and Allergy ,Antibody-Producing Cells ,Gene ,Transcription factor ,Glycoproteins ,030304 developmental biology ,B-Lymphocytes ,0303 health sciences ,Whole Genome Sequencing ,Sequence Analysis, RNA ,PAX5 Transcription Factor ,RNA ,Cell Differentiation ,Original Articles ,Antigens, CD20 ,meta-analysis ,medicine.anatomical_structure ,plasmablast ,PAX5 ,Positive Regulatory Domain I-Binding Factor 1 ,IRF8 ,transcriptome ,030215 immunology - Abstract
International audience; Antibody-secreting cells (ASC) are divided into two principal subsets, including the long-lived plasma cell (PC) subset residing in the bone marrow and the short-lived subset, also called plasmablast (PB). PB are described as a proliferating subset circulating through the blood and ending its differentiation in tissues. Due to their inherent heterogeneity, the molecular signature of PB is not fully established. The purpose of this study was to decipher a specific PB signature in humans and mice through a comprehensive meta-analysis of different data sets exploring the PB differentiation in both species and across different experimental conditions. The present study used recent analyses using whole RNA sequencing in prdm1-GFP transgenic mice to define a reliable and accurate PB signature. Next, we performed similar analysis using current data sets obtained from human PB and PC. The PB-specific signature is composed of 155 and 113 genes in mouse and human being, respectively. Although only nine genes are shared between the human and mice PB signature, the loss of B-cell identity such as the down-regulation of PAX5, MS4A1, (CD20) CD22 and IL-4R is a conserved feature across species and across the different experimental conditions. Additionally, we observed that the IRF8 and IRF4 transcription factors have a specific dynamic range of expression in human PB. We thus demonstrated that IRF4/IRF8 intranuclear staining was useful to define PB in vivo and in vitro and able to discriminate between atypical PB populations and transient states.
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- 2021
19. Molecular Mechanisms Driving IL-10- Producing B Cells Functions: STAT3 and c-MAF as Underestimated Central Key Regulators?
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Magalie Michée-Cospolite, Marina Boudigou, Alexis Grasseau, Quentin Simon, Olivier Mignen, Jacques-Olivier Pers, Divi Cornec, Laëtitia Le Pottier, and Sophie Hillion
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B-Lymphocytes, Regulatory ,Mice ,Proto-Oncogene Proteins c-maf ,Immunology ,Plasma Cells ,Immunology and Allergy ,Animals ,Lymphocyte Count ,Autoimmune Diseases ,Interleukin-10 - Abstract
Regulatory B cells (Bregs) have been highlighted in very different pathology settings including autoimmune diseases, allergy, graft rejection, and cancer. Improving tools for the characterization of Bregs has become the main objective especially in humans. Transitional, mature B cells and plasma cells can differentiate into IL-10 producing Bregs in both mice and humans, suggesting that Bregs are not derived from unique precursors but may arise from different competent progenitors at unrestricted development stages. Moreover, in addition to IL-10 production, regulatory B cells used a broad range of suppressing mechanisms to modulate the immune response. Although Bregs have been consistently described in the literature, only a few reports described the molecular aspects that control the acquisition of the regulatory function. In this manuscript, we detailed the latest reports describing the control of IL-10, TGFβ, and GZMB production in different Breg subsets at the molecular level. We focused on the understanding of the role of the transcription factors STAT3 and c-MAF in controlling IL-10 production in murine and human B cells and how these factors may represent an important crossroad of several key drivers of the Breg response. Finally, we provided original data supporting the evidence that MAF is expressed in human IL-10- producing plasmablast and could be inducedin vitrofollowing different stimulation cocktails. At steady state, we reported that MAF is expressed in specific human B-cell tonsillar subsets including the IgD+CD27+unswitched population, germinal center cells and plasmablast.
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- 2021
20. Synthetic alkyl-ether-lipid promotes TRPV2 channel trafficking trough PI3K/Akt-girdin axis in cancer cells and increases mammary tumour volume
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Audrey Gambade, Marie Potier-Cartereau, Yann Fourbon, Philippe Bougnoux, Romain Félix, Sana Kouba, David Ternant, François Carreaux, Maxime Guéguinou, Ana Bouchet, Paul-Alain Jaffrès, Thomas Harnois, Christophe Vandier, Olivier Mignen, Aurélie Chantôme, Christophe Arnoult, Jean-Pierre Haelters, Thibauld Oullier, Séverine Marionneau-Lambot, Aubin Penna, Günther Weber, Gaëlle Simon, Nutrition, croissance et cancer (U 1069) (N2C), Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Groupe innovation et ciblage cellulaire (GICC), EA 7501 [2018-...] (GICC EA 7501), Université de Tours (UT), Institut de Recherche en Santé de l'Université de Nantes (IRS-UN), The Enteric Nervous System in gut and brain disorders [U1235] (TENS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN), Signalisation et Transports Ioniques Membranaires (STIM), Université de Poitiers-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), Université de Brest (UBO), Chimie, Electrochimie Moléculaires et Chimie Analytique (CEMCA), Université de Brest (UBO)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut Brestois Santé Agro Matière (IBSAM), Institut des Sciences Chimiques de Rennes (ISCR), Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Inserm UMR 1227 Immunothérapies et Pathologies Lymphocytaires B, Hôpital Morvan - CHRU de Brest (CHU - BREST ), INCa, Ligue Nationale Contre le Cancer, Region Centre Val de Loire (LIPIDS project of ARD2020-Biomédicaments), INSERM, Cancéropôle Grand Ouest, the association 'CANCEN' and Tours Hospital oncology association 'ACORT'. M. Guéguinou, Y. Fourbon and A. Gambade held fellowships from the 'Region Centre', 'Ministère de l'Enseignement Supérieur et de la Recherche'., Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA), Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Tours, and Université de Poitiers-Université de Tours-Centre National de la Recherche Scientifique (CNRS)
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0301 basic medicine ,Physiology ,[SDV]Life Sciences [q-bio] ,Metastasis ,03 medical and health sciences ,Transient receptor potential channel ,chemistry.chemical_compound ,0302 clinical medicine ,medicine ,[CHIM]Chemical Sciences ,Molecular Biology ,Protein kinase B ,PI3K/AKT/mTOR pathway ,Constitutive Ca2+ entry ,Chemistry ,[CHIM.ORGA]Chemical Sciences/Organic chemistry ,Breast cancer cell migration ,Cancer ,Cell migration ,Cell Biology ,medicine.disease ,3. Good health ,Cell biology ,TRPV2 ,030104 developmental biology ,Cancer cell ,lipids (amino acids, peptides, and proteins) ,Alkyl-ether-lipid ,030217 neurology & neurosurgery ,Edelfosine - Abstract
International audience; The Transient Receptor Potential Vanilloid type 2 (TRPV2) channel is highly selective for Ca2+ and can be activated by lipids, such as LysoPhosphatidylCholine (LPC). LPC analogues, such as the synthetic alkyl-ether-lipid edelfosine or the endogenous alkyl-ether-lipid Platelet Activating Factor (PAF), modulates ion channels in cancer cells. This opens the way to develop alkyl-ether-lipids for the modulation of TRPV2 in cancer. Here, we investigated the role of 2-Acetamido-2-Deoxy-l-O-Hexadecyl-rac-Glycero-3-PhosphatidylCholine (AD-HGPC), a new alkyl-ether-lipid (LPC analogue), on TRPV2 trafficking and its impact on Ca2+ -dependent cell migration. The effect of AD-HGPC on the TRPV2 channel and tumour process was further investigated using calcium imaging and an in vivo mouse model. Using molecular and pharmacological approaches, we dissected the mechanism implicated in alkyl-ether-lipids sensitive TRPV2 trafficking. We found that TRPV2 promotes constitutive Ca2+ entry, leading to migration of highly metastatic breast cancer cell lines through the PI3K/Akt-Girdin axis. AD-HGPC addresses the functional TRPV2 channel in the plasma membrane through Golgi stimulation and PI3K/Akt/Rac-dependent cytoskeletal reorganization, leading to constitutive Ca2+ entry and breast cancer cell migration (without affecting the development of metastasis), in a mouse model. We describe, for the first time, the biological role of a new alkyl-ether-lipid on TRPV2 channel trafficking in breast cancer cells and highlight the potential modulation of TRPV2 by alkyl-ether-lipids as a novel avenue for research in the treatment of metastatic cancer.
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- 2021
21. Pacific-Ciguatoxin-2 and Brevetoxin-1 Induce the Sensitization of Sensory Receptors Mediating Pain and Pruritus in Sensory Neurons
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Joachim W. Fluhr, Richard J. Lewis, Laurent Misery, Raphaele Le Garrec, Raphael Leschiera, Nelig Le Goux, Olivier Mignen, Maxime Fouchard, Ophélie Pierre, and Paul Buscaglia
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0301 basic medicine ,Aquatic Organisms ,Ciguatoxin ,Sensory Receptor Cells ,QH301-705.5 ,receptors ,Pain ,Pharmaceutical Science ,Sensory system ,ciguatera fish poisoning ,Pharmacology ,Article ,sensitization ,Ciguatoxins ,03 medical and health sciences ,Brevetoxin ,0302 clinical medicine ,Neurotrophic factors ,Drug Discovery ,medicine ,Animals ,Rats, Wistar ,neurotoxic shellfish poisoning ,Biology (General) ,Receptor ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Sensitization ,sensory disorders ,Pacific Ocean ,biology ,business.industry ,Oxocins ,ion channels ,Ciguatera Poisoning ,pruritus ,Rats ,030104 developmental biology ,medicine.anatomical_structure ,Nerve growth factor ,Animals, Newborn ,Models, Animal ,biology.protein ,Marine Toxins ,business ,ciguatoxin ,030217 neurology & neurosurgery ,Neurotrophin ,brevetoxin - Abstract
Ciguatera fish poisoning (CFP) and neurotoxic shellfish poisoning syndromes are induced by the consumption of seafood contaminated by ciguatoxins and brevetoxins. Both toxins cause sensory symptoms such as paresthesia, cold dysesthesia and painful disorders. An intense pruritus, which may become chronic, occurs also in CFP. No curative treatment is available and the pathophysiology is not fully elucidated. Here we conducted single-cell calcium video-imaging experiments in sensory neurons from newborn rats to study in vitro the ability of Pacific-ciguatoxin-2 (P-CTX-2) and brevetoxin-1 (PbTx-1) to sensitize receptors and ion channels, (i.e., to increase the percentage of responding cells and/or the response amplitude to their pharmacological agonists). In addition, we studied the neurotrophin release in sensory neurons co-cultured with keratinocytes after exposure to P-CTX-2. Our results show that P-CTX-2 induced the sensitization of TRPA1, TRPV4, PAR2, MrgprC, MrgprA and TTX-r NaV channels in sensory neurons. P-CTX-2 increased the release of nerve growth factor and brain-derived neurotrophic factor in the co-culture supernatant, suggesting that those neurotrophins could contribute to the sensitization of the aforementioned receptors and channels. Our results suggest the potential role of sensitization of sensory receptors/ion channels in the induction or persistence of sensory disturbances in CFP syndrome.
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- 2021
22. IL-21 and IFN-alpha have both opposite and redundant role on human innate precursors and memory B-cell differentiation
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Marina Boudigou, Magalie Michée-Cospolite, Patrice Hémon, Alexis Grasseau, Christelle Le Dantec, Emmanuelle Porchet, Christophe Jamin, Valérie Devauchelle, Olivier Mignen, Divi Cornec, Jacques-Olivier Pers, Laëtitia Le Pottier, and Sophie Hillion
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education.field_of_study ,biology ,Population ,Interleukin ,Spleen ,Marginal zone ,Immunoglobulin D ,In vitro ,Cell biology ,medicine.anatomical_structure ,Antigen ,Apoptosis ,biology.protein ,medicine ,education - Abstract
Immunological memory is essential for effective immune protection upon antigen rechallenge. Memory B cells encompass multiple subsets, heterogeneous in terms of phenotypes, origins and precursors, anatomical localization, and functional responses. B-cell responses are conditioned by micro-environmental signals, including cytokines. Here, we analyzed in vitro the effects of two cytokines implicated in B-cell differentiation, interferon-alpha (IFN-α) and interleukin (IL)-21, on the early functional response of four different mature B-cell subsets (IgD- CD27- naive, IgD+ CD27+ unswitched, IgD- CD27+ switched and double-negative B cells). The dual response of naive and memory B cells to IL-21 allowed us to uncover a unique IgD+ CD27- CD10- B-cell population (referred to as NARB+) characterized by the expression of marginal zone B-cell markers CD45RB and CD1c. Similar to memory B cells, NARB+ cells were in a pre-activated state, allowing them to rapidly differentiate into plasmablasts upon innate signals while maintaining their susceptibility to IL-21 activation-induced apoptosis as observed for the naive compartment. Both in-depth phenotypic analysis of circulating B cells, and identification of these cells in spleen, tonsil and gut-associated lymphoid tissues, supported that NARB+ are uncommitted precursors of human marginal zone B cells.
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- 2021
23. The p.E152K-STIM1 mutation deregulates Ca2+ signaling contributing to chronic pancreatitis
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Thierry Capiod, Emmanuelle Masson, Wesley H. Brooks, Reginald Philippe, Nicolas Lebonvallet, Wayne C. Guida, Pauline Dubar, Miguel Burgos, Juan Llopis, Fabrice Antigny, Olivier Mignen, Peter B. Stathopulos, Claude Férec, Mitsuhiko Ikura, Maud Frieden, F. Campeotto, Beatriz Domingo, Sreya Mukherjee, Cédric Le Maréchal, Paul Buscaglia, Jian-Min Chen, Laboratoire sur les interactions Epithéliums Neurones (LIEN), and Université de Brest (UBO)
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0303 health sciences ,Stromal cell ,SERCA ,[SDV]Life Sciences [q-bio] ,Endoplasmic reticulum ,STIM1 ,Cell Biology ,Biology ,medicine.disease ,Cell biology ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Pancreatitis ,Secretion ,ddc:612 ,030217 neurology & neurosurgery ,Homeostasis ,Intracellular ,030304 developmental biology - Abstract
Since deregulation of intracellular Ca2+ can lead to intracellular trypsin activation, and stromal interaction molecule-1 (STIM1) protein is the main regulator of Ca2+ homeostasis in pancreatic acinar cells, we explored the Ca2+ signaling in 37 STIM1 variants found in three pancreatitis patient cohorts. Extensive functional analysis of one particular variant, p.E152K, identified in three patients, provided a plausible link between dysregulated Ca2+ signaling within pancreatic acinar cells and chronic pancreatitis susceptibility. Specifically, p.E152K, located within the STIM1 EF-hand and sterile α-motif domain, increased the release of Ca2+ from the endoplasmic reticulum in patient-derived fibroblasts and transfected HEK293T cells. This event was mediated by altered STIM1–sarco/endoplasmic reticulum calcium transport ATPase (SERCA) conformational change and enhanced SERCA pump activity leading to increased store-operated Ca2+ entry (SOCE). In pancreatic AR42J cells expressing the p.E152K variant, Ca2+ signaling perturbations correlated with defects in trypsin activation and secretion, and increased cytotoxicity after cholecystokinin stimulation. This article has an associated First Person interview with the first author of the paper.
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- 2021
24. Dual Response to IL-21 and IFN-Alpha Reveals Human B-Cell Precursors With Multiple Differentiation Potentials
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Christophe Jamin, Christelle Le Dantec, Magalie Michée-Cospolite, Divi Cornec, Marina Boudigou, Valérie Devauchelle, Alexis Grasseau, Patrice Hemon, Olivier Mignen, Jacques-Olivier Pers, Emanuelle Porchet, Sophie Hillion, and Laëtitia Le Pottier
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education.field_of_study ,Population ,Interleukin ,chemical and pharmacologic phenomena ,Spleen ,Biology ,Marginal zone ,Immunoglobulin D ,In vitro ,Cell biology ,medicine.anatomical_structure ,Antigen ,Apoptosis ,medicine ,biology.protein ,education - Abstract
Immunological memory is essential for effective immune protection upon antigen rechallenge. Memory B cells encompass multiple subsets, heterogeneous in terms of phenotypes, origins and precursors, anatomical localization, and functional responses. B-cell responses are conditioned by micro-environmental signals, including cytokines. Here, we analyzed in vitro the effects of two cytokines implicated in B-cell differentiation, interferon-alpha (IFN-α) and interleukin (IL)-21, on the early functional response of four different mature B-cell subsets (IgD+ CD27- naive, IgD+ CD27+ unswitched, IgD- CD27+ switched and double-negative B cells). The dual response of naive and memory B cells to IL-21 allowed us to uncover a unique IgD+ CD27- CD10- B-cell population characterized by the expression of marginal zone B-cell markers CD45RB and CD1c (referred to as NARB+). Similar to memory B cells, NARB+ cells displayed a pre-activated state, allowing them to rapidly differentiate into plasmablasts upon innate signals. However, this population also maintained its susceptibility to IL-21 activation-induced apoptosis similarly to the naive compartment. Both in-depth phenotypic analysis of circulating B cells, and identification of these cells in spleen, tonsil, and gut-associated lymphoid tissues, suggested that NARB+ could be uncommitted precursors of different developmental pathways including human marginal zone-like B cells (MZB). In this regard, we showed that patients with primary Sjogren’s syndrome exhibited a severe reduction of IgD+CD27+ MZB-like cells associated with a diminution of the NARB+ subset suggesting a developmental relationship.
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- 2021
25. Calcium Increase and Substance P Release Induced by the Neurotoxin Brevetoxin-1 in Sensory Neurons: Involvement of PAR2 Activation through Both Cathepsin S and Canonical Signaling
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Paul Buscaglia, Raphael Leschiera, Olivier Mignen, Nelig Le Goux, Joachim W. Fluhr, Ophélie Pierre, Raphaele Le Garrec, Laurent Misery, and Maxime Fouchard
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0301 basic medicine ,Keratinocytes ,Sensory Receptor Cells ,substance P ,TRPV Cation Channels ,canonical pathway ,Article ,03 medical and health sciences ,Transient receptor potential channel ,Brevetoxin ,0302 clinical medicine ,Calcium imaging ,Neurotoxin ,Animals ,Humans ,Receptor, PAR-2 ,biased pathway ,Rats, Wistar ,Protein kinase A ,Evoked Potentials ,lcsh:QH301-705.5 ,Protein kinase C ,Cells, Cultured ,Protein Kinase C ,sensory disorders ,Chemistry ,Oxocins ,General Medicine ,Dipeptides ,Isoxazoles ,Inositol trisphosphate receptor ,Cathepsins ,PAR2 ,Recombinant Proteins ,Cell biology ,Rats ,030104 developmental biology ,lcsh:Biology (General) ,cathepsin S ,Calcium ,Marine Toxins ,Signal transduction ,ciguatoxin ,030217 neurology & neurosurgery ,Signal Transduction ,brevetoxin - Abstract
Red tides involving Karenia brevis expose humans to brevetoxins (PbTxs). Oral exposition triggers neurotoxic shellfish poisoning, whereas inhalation induces a respiratory syndrome and sensory disturbances. No curative treatment is available and the pathophysiology is not fully elucidated. Protease-activated receptor 2 (PAR2), cathepsin S (Cat-S) and substance P (SP) release are crucial mediators of the sensory effects of ciguatoxins (CTXs) which are PbTx analogs. This work explored the role of PAR2 and Cat-S in PbTx-1-induced sensory effects and deciphered the signaling pathway involved. We performed calcium imaging, PAR2 immunolocalization and SP release experiments in monocultured sensory neurons or co-cultured with keratinocytes treated with PbTx-1 or P-CTX-2. We demonstrated that PbTx-1-induced calcium increase and SP release involved Cat-S, PAR2 and transient receptor potential vanilloid 4 (TRPV4). The PbTx-1-induced signaling pathway included protein kinase A (PKA) and TRPV4, which are compatible with the PAR2 biased signaling induced by Cat-S. Internalization of PAR2 and protein kinase C (PKC), inositol triphosphate receptor and TRPV4 activation evoked by PbTx-1 are compatible with the PAR2 canonical signaling. Our results suggest that PbTx-1-induced sensory disturbances involve the PAR2-TRPV4 pathway. We identified PAR2, Cat-S, PKA, and PKC that are involved in TRPV4 sensitization induced by PbTx-1 in sensory neurons.
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- 2020
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26. Keratinocytes Communicate with Sensory Neurons via Synaptic‐like Contacts
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Killian L’Herondelle, Laurent Misery, Laëtitia Le Pottier, Denis Ressnikoff, Nathalie Kerfant, Olivier Mignen, Jean-Pierre Pennec, Philippe Elies, Raphaele Le Garrec, Matthieu Talagas, Marek Haftek, Alexia Reux, Raphael Leschiera, Nicolas Lebonvallet, Veronica La Padula, Gerard Sinquin, Pascale Marcorelles, Laboratoire sur les interactions Epithéliums Neurones (LIEN), and Université de Brest (UBO)
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Adult ,Keratinocytes ,Male ,0301 basic medicine ,Sensory Receptor Cells ,[SDV]Life Sciences [q-bio] ,Synaptophysin ,Human skin ,Sensory system ,Biology ,Synaptic vesicle ,Synaptotagmin 1 ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Humans ,ComputingMilieux_MISCELLANEOUS ,Aged ,Qa-SNARE Proteins ,Middle Aged ,Coculture Techniques ,Rats ,Microscopy, Electron ,030104 developmental biology ,medicine.anatomical_structure ,Neurology ,Synaptotagmin I ,Synapses ,biology.protein ,Female ,Synaptic Vesicles ,Neurology (clinical) ,Neuron ,Epidermis ,Keratinocyte ,Neuroscience ,Free nerve ending ,030217 neurology & neurosurgery - Abstract
Objective Pain, temperature, and itch are conventionally thought to be exclusively transduced by the intraepidermal nerve endings. Although recent studies have shown that epidermal keratinocytes also participate in sensory transduction, the mechanism underlying keratinocyte communication with intraepidermal nerve endings remains poorly understood. We sought to demonstrate the synaptic character of the contacts between keratinocytes and sensory neurons and their involvement in sensory communication between keratinocytes and sensory neurons. Methods Contacts were explored by morphological, molecular, and functional approaches in cocultures of epidermal keratinocytes and sensory neurons. To interrogate whether structures observed in vitro were also present in the human epidermis, in situ correlative light electron microscopy was performed on human skin biopsies. Results Epidermal keratinocytes dialogue with sensory neurons through en passant synaptic-like contacts. These contacts have the ultrastructural features and molecular hallmarks of chemical synaptic-like contacts: narrow intercellular cleft, keratinocyte synaptic vesicles expressing synaptophysin and synaptotagmin 1, and sensory information transmitted from keratinocytes to sensory neurons through SNARE-mediated (syntaxin1) vesicle release. Interpretation By providing selective communication between keratinocytes and sensory neurons, synaptic-like contacts are the hubs of a 2-site receptor. The permanent epidermal turnover, implying a specific en passant structure and high plasticity, may have delayed their identification, thereby contributing to the long-held concept of nerve endings passing freely between keratinocytes. The discovery of keratinocyte-sensory neuron synaptic-like contacts may call for a reassessment of basic assumptions in cutaneous sensory perception and sheds new light on the pathophysiology of pain and itch as well as the physiology of touch. ANN NEUROL 2020;88:1205-1219.
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- 2020
27. CD5 and B lymphocyte responses: multifaceted effects through multitudes of pathways and channels
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Jacques-Olivier Pers, Yves Renaudineau, Rizgar A. Mageed, Olivier Mignen, Taher E. Taher, Jonas Bystrom, Insitute of Immunology and Immunotherapy, Birmingham (University of Birmingham), Centre for Experimental Medicine and Rheumatology (William Harvey Research Institute), Canalopathy and Calcium Signaling, INSERM UMR1078, Brest, France, Immunologie et Pathologie (EA 2216), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire d'Immunologie et Immunothérapie [Brest], and Centre Hospitalier Régional Universitaire de Brest (CHRU Brest)
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Cell Survival ,Lymphocyte ,T-Lymphocytes ,[SDV]Life Sciences [q-bio] ,Immunology ,Biology ,CD5 Antigens ,Models, Biological ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,Correspondence ,medicine ,Immunology and Allergy ,Humans ,ComputingMilieux_MISCELLANEOUS ,030304 developmental biology ,0303 health sciences ,B-Lymphocytes ,Interleukin-10 ,Infectious Diseases ,medicine.anatomical_structure ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,CD5 ,030215 immunology ,Signal Transduction - Abstract
International audience
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- 2020
28. Potassium and Calcium Channel Complexes as Novel Targets for Cancer Research
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Marie, Potier-Cartereau, William, Raoul, Gunther, Weber, Karine, Mahéo, Raphael, Rapetti-Mauss, Maxime, Gueguinou, Paul, Buscaglia, Caroline, Goupille, Nelig, Le Goux, Souleymane, Abdoul-Azize, Thierry, Lecomte, Gaëlle, Fromont, Aurélie, Chantome, Olivier, Mignen, Olivier, Soriani, and Christophe, Vandier
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Potassium Channels ,Potassium Channels, Voltage-Gated ,Neoplasms ,Potassium ,Humans ,Calcium ,Calcium Channels ,Lipids - Abstract
The intracellular Ca
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- 2020
29. The p.E152K-STIM1 mutation deregulates Ca
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Miguel, Burgos, Reginald, Philippe, Fabrice, Antigny, Paul, Buscaglia, Emmanuelle, Masson, Sreya, Mukherjee, Pauline, Dubar, Cédric, Le Maréchal, Florence, Campeotto, Nicolas, Lebonvallet, Maud, Frieden, Juan, Llopis, Beatriz, Domingo, Peter B, Stathopulos, Mitsuhiko, Ikura, Wesley, Brooks, Wayne, Guida, Jian-Min, Chen, Claude, Ferec, Thierry, Capiod, and Olivier, Mignen
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HEK293 Cells ,ORAI1 Protein ,Pancreatitis, Chronic ,Mutation ,Humans ,Calcium ,Calcium Signaling ,Stromal Interaction Molecule 1 ,Endoplasmic Reticulum ,Neoplasm Proteins - Abstract
Since deregulation of intracellular Ca
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- 2020
30. p.E152K-STIM1 mutation deregulates Ca2+ signaling contributing to chronic pancreatitis
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Wesley H. Brooks, Juan Llopis, Olivier Mignen, Mitsuhiko Ikura, Fabrice Antigny, Emmanuelle Masson, Paul Buscaglia, Reginald Philippe, Sreya Mukherjee, F. Campeotto, Cédric Le Maréchal, Maud Frieden, Beatriz Domingo, Jian-Min Chen, Nicolas Lebonvallet, Wayne C. Guida, Peter B. Stathopulos, Claude Férec, Thierry Capiod, Pauline Dubar, and Miguel Burgos
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inorganic chemicals ,Calcium metabolism ,0303 health sciences ,SERCA ,Chemistry ,Endoplasmic reticulum ,STIM1 ,medicine.disease ,Store-operated calcium entry ,Cell biology ,03 medical and health sciences ,0302 clinical medicine ,Acinar cell ,medicine ,Pancreatitis ,030217 neurology & neurosurgery ,Intracellular ,030304 developmental biology - Abstract
Since deregulation of intracellular Ca2+ can lead to intracellular trypsin activation and STIM1 (stromal interaction molecule-1) protein is the main regulator of Ca2+ homeostasis in pancreatic acinar cells, we explored the Ca2+ signaling in 37 STIM1 variants found in three pancreatitis patient cohorts. Extensive functional analysis of one particular variant, p.E152K, identified in three patients, provided a plausible link between dysregulated Ca2+ signaling within pancreatic acinar cells and chronic pancreatitis susceptibility. Specifically, p.E152K, located within the STIM1 EF-hand and sterile α-motif domain, increased the release of Ca2+ from the endoplasmic reticulum in patient-derived fibroblasts and transfected HEK293T cells. This event was mediated by altered STIM1-sarco/endoplasmic reticulum calcium transport ATPase (SERCA) interactions and enhanced SERCA pump activity leading to increased Store Operated Calcium Entry (SOCE). In the pancreatic AR42J cells expressing the p.E152K variant, Ca2+-signaling perturbations correlated with defects in trypsin activation and secretion, and increased cytotoxicity after cholecystokinin stimulation.Summary statementp.E152K-STIM1 variant found in pancreatitis patients leads to intracellular changes in calcium homeostasis through SERCA interaction, enabling intracellular trypsin activation and pancreatic acinar cell death.
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- 2020
31. Potassium and Calcium Channel Complexes as Novel Targets for Cancer Research
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Karine Mahéo, Nelig Le Goux, Günther Weber, Aurélie Chantôme, Paul Buscaglia, Thierry Lecomte, Olivier Soriani, Marie Potier-Cartereau, Olivier Mignen, Raphael Rapetti-Mauss, Souleymane Abdoul-Azize, Caroline Goupille, Maxime Guéguinou, William Raoul, Christophe Vandier, and Gaëlle Fromont
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0303 health sciences ,Chemistry ,Calcium channel ,Depolarization ,Membrane hyperpolarization ,3. Good health ,03 medical and health sciences ,Cytosol ,0302 clinical medicine ,Membrane ,030220 oncology & carcinogenesis ,Cancer research ,Receptor ,Flux (metabolism) ,Intracellular ,030304 developmental biology - Abstract
The intracellular Ca2+ concentration is mainly controlled by Ca2+ channels. These channels form complexes with K+ channels, which function to amplify Ca2+ flux. In cancer cells, voltage-gated/voltage-dependent Ca2+ channels and non-voltage-gated/voltage-independent Ca2+ channels have been reported to interact with K+ channels such as Ca2+-activated K+ channels and voltage-gated K+ channels. These channels are activated by an increase in cytosolic Ca2+ concentration or by membrane depolarization, which induces membrane hyperpolarization, increasing the driving force for Ca2+ flux. These complexes, composed of K+ and Ca2+ channels, are regulated by several molecules including lipids (ether lipids and cholesterol), proteins (e.g. STIM), receptors (e.g. S1R/SIGMAR1), and peptides (e.g. LL-37) and can be targeted by monoclonal antibodies, making them novel targets for cancer research.
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- 2020
32. STIM1 at the plasma membrane as a new target in progressive chronic lymphocytic leukemia
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Nelig Le Goux, Marie Potier-Cartereau, Olivier Mignen, Adrian Tempescul, Tinhinane Fali, Sarra Melayah, Yves Renaudineau, Patrice Hemon, Cristina Bagacean, Christophe Vandier, Paul Buscaglia, Marjolaine Debant, Miguel Burgos, Christian Berthou, Jacques-Olivier Pers, Lymphocyte B et Auto-immunité (LBAI), Université de Brest (UBO)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire d'Immunologie et Immunothérapie [Brest], Centre Hospitalier Régional Universitaire de Brest (CHRU Brest), Nutrition, croissance et cancer (U 1069) (N2C), Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM), Cancéropôle Grand-Ouest, CHRU Brest - Service d'Hématologie (CHU-Brest-Hemato), Laboratoire d'Immunologie et Immunothérapie, Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Michel, Geneviève, Lymphocytes B, Autoimmunité et Immunothérapies (LBAI), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-LabEX IGO Immunothérapie Grand Ouest, Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ)-Institut Brestois Santé Agro Matière (IBSAM), and Université de Brest (UBO)
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Male ,0301 basic medicine ,Cancer Research ,ORAI1 Protein ,Chronic lymphocytic leukemia ,[SDV]Life Sciences [q-bio] ,chemistry.chemical_compound ,Antineoplastic Agents, Immunological ,0302 clinical medicine ,hemic and lymphatic diseases ,Tumor Cells, Cultured ,Immunology and Allergy ,Prospective Studies ,RNA, Small Interfering ,TRPC ,B-Lymphocytes ,Constitutive Ca2+ entry ,Voltage-dependent calcium channel ,Chemistry ,breakpoint cluster region ,Antibodies, Monoclonal ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Neoplasm Proteins ,[SDV] Life Sciences [q-bio] ,Treatment Outcome ,medicine.anatomical_structure ,Oncology ,Gene Knockdown Techniques ,030220 oncology & carcinogenesis ,Disease Progression ,Molecular Medicine ,Female ,Signal transduction ,Research Article ,Thapsigargin ,Cell Survival ,STIM1 ,Primary Cell Culture ,Immunology ,B-cell receptor ,lcsh:RC254-282 ,03 medical and health sciences ,medicine ,Humans ,Calcium Signaling ,Stromal Interaction Molecule 1 ,Disease outcome ,B cell ,Aged ,TRPC Cation Channels ,Pharmacology ,Cell Membrane ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,030104 developmental biology ,Cancer research ,Calcium ,CLL - Abstract
Background Dysregulation in calcium (Ca2+) signaling is a hallmark of chronic lymphocytic leukemia (CLL). While the role of the B cell receptor (BCR) Ca2+ pathway has been associated with disease progression, the importance of the newly described constitutive Ca2+ entry (CE) pathway is less clear. In addition, we hypothesized that these differences reflect modifications of the CE pathway and Ca2+ actors such as Orai1, transient receptor potential canonical (TRPC) 1, and stromal interaction molecule 1 (STIM1), the latter being the focus of this study. Methods An extensive analysis of the Ca2+ entry (CE) pathway in CLL B cells was performed including constitutive Ca2+ entry, basal Ca2+ levels, and store operated Ca2+ entry (SOCE) activated following B cell receptor engagement or using Thapsigargin. The molecular characterization of the calcium channels Orai1 and TRPC1 and to their partner STIM1 was performed by flow cytometry and/or Western blotting. Specific siRNAs for Orai1, TRPC1 and STIM1 plus the Orai1 channel blocker Synta66 were used. CLL B cell viability was tested in the presence of an anti-STIM1 monoclonal antibody (mAb, clone GOK) coupled or not with an anti-CD20 mAb, rituximab. The Cox regression model was used to determine the optimal threshold and to stratify patients. Results Seeking to explore the CE pathway, we found in untreated CLL patients that an abnormal CE pathway was (i) highly associated with the disease outcome; (ii) positively correlated with basal Ca2+ concentrations; (iii) independent from the BCR-PLCγ2-InsP3R (SOCE) Ca2+ signaling pathway; (iv) supported by Orai1 and TRPC1 channels; (v) regulated by the pool of STIM1 located in the plasma membrane (STIM1PM); and (vi) blocked when using a mAb targeting STIM1PM. Next, we further established an association between an elevated expression of STIM1PM and clinical outcome. In addition, combining an anti-STIM1 mAb with rituximab significantly reduced in vitro CLL B cell viability within the high STIM1PM CLL subgroup. Conclusions These data establish the critical role of a newly discovered BCR independent Ca2+ entry in CLL evolution, provide new insights into CLL pathophysiology, and support innovative therapeutic perspectives such as targeting STIM1 located at the plasma membrane. Electronic supplementary material The online version of this article (10.1186/s40425-019-0591-3) contains supplementary material, which is available to authorized users.
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- 2019
33. CD5 expression promotes IL-10 production through activation of the MAPK/Erk pathway and upregulation of TRPC1 channels in B lymphocytes
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Sarra Melayah, Marjolaine Debant, Miguel Burgos, Soizic Garaud, Olivier Mignen, Kaushal Parikh, Pierre Youinou, Maikel P. Peppelenbosch, Damien Luque-Paz, David A. Isenberg, Yves Renaudineau, Jacques-Olivier Pers, Rizgar A. Mageed, Gilles Chiocchia, Taher E. Taher, Christian Berthou, Michel, Geneviève, Clinical Chemistry, Gastroenterology & Hepatology, Other departments, Center of Experimental and Molecular Medicine, Immunologie et Pathologie (EA2216), Université de Brest (UBO)-IFR148, William Harvey Research Institute, Barts and the London Medical School, Pontifical Catholic University of Rio de Janeiro (PUC), Centre for Experimental and Molecular Medicine, Centre For Experimental and Molecular Medicine, Laboratoire d'Immunologie et Immunothérapie, Centre Hospitalier Régional Universitaire de Brest (CHRU Brest), Physiopathologie et Pharmacologie Clinique de la Douleur (LPPD), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Institut National de la Santé et de la Recherche Médicale (INSERM), Department of Gastroenterology and Hepatology, Erasmus University Medical Center [Rotterdam] (Erasmus MC), and Centre for Rheumatology - London
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0301 basic medicine ,MAPK/ERK pathway ,Male ,Small interfering RNA ,[SDV.IMM] Life Sciences [q-bio]/Immunology ,MAP Kinase Signaling System ,Immunology ,Receptors, Antigen, B-Cell ,Biology ,CD5 Antigens ,Models, Biological ,03 medical and health sciences ,Downregulation and upregulation ,immune system diseases ,hemic and lymphatic diseases ,Cell Line, Tumor ,medicine ,Immunology and Allergy ,Humans ,Phosphorylation ,B cell ,ComputingMilieux_MISCELLANEOUS ,Aged ,TRPC Cation Channels ,Aged, 80 and over ,B-Lymphocytes ,breakpoint cluster region ,Middle Aged ,Leukemia, Lymphocytic, Chronic, B-Cell ,3. Good health ,Cell biology ,Interleukin-10 ,Up-Regulation ,Interleukin 10 ,030104 developmental biology ,Infectious Diseases ,medicine.anatomical_structure ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,Calcium ,Female ,CD5 ,Signal transduction ,Transcriptome ,Research Article - Abstract
CD5 is constitutively expressed on T cells and a subset of mature normal and leukemic B cells in patients with chronic lymphocytic leukemia (CLL). Important functional properties are associated with CD5 expression in B cells, including signal transducer and activator of transcription 3 activation, IL-10 production and the promotion of B-lymphocyte survival and transformation. However, the pathway(s) by which CD5 influences the biology of B cells and its dependence on B-cell receptor (BCR) co-signaling remain unknown. In this study, we show that CD5 expression activates a number of important signaling pathways, including Erk1/2, leading to IL-10 production through a novel pathway independent of BCR engagement. This pathway is dependent on extracellular calcium (Ca2+) entry facilitated by upregulation of the transient receptor potential channel 1 (TRPC1) protein. We also show that Erk1/2 activation in a subgroup of CLL patients is associated with TRPC1 overexpression. In this subgroup of CLL patients, small inhibitory RNA (siRNA) for CD5 reduces TRPC1 expression. Furthermore, siRNAs for CD5 or for TRPC1 inhibit IL-10 production. These findings provide new insights into the role of CD5 in B-cell biology in health and disease and could pave the way for new treatment strategies for patients with B-CLL.
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- 2018
34. Les kératinocytes épidermiques communiquent avec les neurones sensoriels par l’intermédiaire de contacts de type synaptique
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Laurent Misery, Olivier Mignen, Denis Ressnikoff, Jean-Pierre Pennec, Gerard Sinquin, Laetitia Le Pottier, Raphael Leschiera, Killian L’Herondelle, Philippe Elies, N. Kerfant, Marek Haftek, Alexia Reux, Matthieu Talagas, Pascale Marcorelles, Raphaele Le Garrec, Nicolas Lebonvallet, and Veronica La Padula
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Anatomy - Abstract
La temperature, la douleur et le prurit sont classiquement considerees comme etant exclusivement percus par les fibres nerveuses intra-epidermiques. Alors que des etudes recentes ont demontre que les keratinocytes epidermiques participent egalement a la transduction sensorielle cutanee, le mecanisme qui sous-tend la communication entre les keratinocytes et les fibres nerveuses intra-epidermiques reste meconnu. Nous demontrons, dans cette etude, que les kerationocytes dialoguent avec les neurones sensoriels par l’intermediaire de contacts « en passant » de type synaptique. Ces contacts ont les caracteristiques ultrastructurales et les caracteristiques moleculaires de synapses chimiques, vehiculant des informations sensorielles des keratinocytes vers les neurones sensoriels par l’intermediaire d’une exocytose mediee par des complexes SNAREs. Le renouvellement permanent de l’epiderme, qui implique une structure specifique de type « en passant » ainsi qu’une grande plasticite, pourrait avoir retarde leur identification, contribuant par consequent a la perennite du concept de fibres nerveuses intra-epidermiques cheminant librement entre les keratinocytes. En assurant une communication selective entre les keratinocytes et les neurones sensoriels, ces contacts synaptiques pourraient etre le pivot d’un recepteur a deux sites, l’un keratinocytaire et l’autre neuronal. La decouverte de ces contacts neuro-keratinocytaires de type synaptique pourrait amener a une reconsideration des concepts etablis au sujet de la perception sensorielle cutanee, et invite a porter un regard nouveau sur la physiopathologie de la douleur et du prurit, aussi bien que de la physiologie du toucher.
- Published
- 2021
35. Dysregulated Lymphoid Cell Populations in Mouse Models of Systemic Lupus Erythematosus
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Aurélie De Groof, Patrice Hemon, Bernard Lauwerys, Yves Renaudineau, Olivier Mignen, Jacques-Olivier Pers, Edward K. Wakeland, European Space Research and Technology Centre (ESTEC), European Space Agency (ESA), Lymphocyte B et Auto-immunité (LBAI), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO), University of Texas Southwestern Medical Center [Dallas], Laboratoire d'Immunologie et Immunothérapie [Brest], Centre Hospitalier Régional Universitaire de Brest (CHRU Brest), and Université Catholique de Louvain = Catholic University of Louvain (UCL)
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0301 basic medicine ,Mice, Inbred MRL lpr ,T-Lymphocytes ,T cell ,Cell ,Autoimmunity ,Biology ,Lymphocyte Activation ,Mouse models ,medicine.disease_cause ,Mice ,03 medical and health sciences ,Systemic lupus erythematosus ,0302 clinical medicine ,medicine ,Animals ,Humans ,Lupus Erythematosus, Systemic ,Immunology and Allergy ,Flow cytometry ,skin and connective tissue diseases ,B cell ,Autoantibodies ,B-Lymphocytes ,Autoantibody ,Peripheral tolerance ,General Medicine ,Chromatin ,Lymphocyte Subsets ,3. Good health ,Disease Models, Animal ,Self Tolerance ,030104 developmental biology ,medicine.anatomical_structure ,Immunology ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,Disease Susceptibility ,Bone marrow ,Immunologic Memory ,030215 immunology - Abstract
International audience; Biases in the distribution and phenotype of T, B, and antigen-presenting cell populations are strongly connected to mechanisms of disease development in mouse models of systemic lupus erythematosus (SLE). Here, we describe longitudinal changes in lymphoid and antigen-presenting cell subsets in bone marrow, blood and spleen from two lupus-prone strains (MRL/lpr and B6.Sle1.Sle2.Sle3 tri-congenic mice), and how they integrate in our present understanding of the pathogenesis of the disease. In particular, we focus on (autoreactive) T cell activation patterns in lupus-prone mice. Break of T cell tolerance to chromatin constituents (histone peptides) is key to the development of the disease and is related to T cell intrinsic defects, contributed by genetic susceptibility factors and by extrinsic amplificatory mechanisms, in particular over-stimulation by antigen-presenting cells. We also describe shifts in B cell sub-populations, going from skewed immature B cell populations as an indication of disturbed central and peripheral tolerance checkpoints, to enriched long-lived plasma cells, which are key to persistent autoantibody production in the disease. B cell activation mechanisms in SLE are both T cell-dependent (break of tolerance and production of specific autoantibodies) and -independent (polyclonal B cell activation, production of autoantibodies by long-lived plasma cells). By providing a comprehensive evaluation of B and T cell surface markers in two major mouse models of SLE and a description of their changes before and after disease onset, this review illustrates how the study of lymphoid cell phenotype delivers key information regarding pathogenic pathways and supplies tools to assess the beneficial effects of novel therapeutic interventions.
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- 2017
36. Molecular Dynamics Simulations of Membrane-Bound STIM1 to Investigate Conformational Changes during STIM1 Activation upon Calcium Release
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Aleksandra Karolak, Paul Buscaglia, Olivier Mignen, Yves Renaudineau, Sreya Mukherjee, Wayne C. Guida, Wesley H. Brooks, Marjolaine Debant, School of Education Technology [Kolkata, West Bengal], Jadavpur University, Department of Chemistry University of South Florida, University of South Florida [Tampa] (USF), Lymphocyte B et Auto-immunité (LBAI), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO), Laboratoire d'Immunologie et Immunothérapie [Brest], Centre Hospitalier Régional Universitaire de Brest (CHRU Brest), and Department of Chemistry, University of South Florida, Tampa, FL, USA.
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inorganic chemicals ,0301 basic medicine ,General Chemical Engineering ,chemistry.chemical_element ,Molecular Dynamics Simulation ,Library and Information Sciences ,Calcium ,Calcium in biology ,Cell membrane ,03 medical and health sciences ,0302 clinical medicine ,Protein Domains ,medicine ,Humans ,Stromal Interaction Molecule 1 ,Calcium signaling ,Voltage-dependent calcium channel ,Chemistry ,Cell Membrane ,STIM1 ,General Chemistry ,Neoplasm Proteins ,Computer Science Applications ,Cell biology ,Cytosol ,030104 developmental biology ,medicine.anatomical_structure ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,030217 neurology & neurosurgery ,Intracellular - Abstract
International audience; Calcium is involved in important intracellular processes, such as intracellular signaling from cell membrane receptors to the nucleus. Typically, calcium levels are kept at less than 100 nM in the nucleus and cytosol, but some calcium is stored in the endoplasmic reticulum (ER) lumen for rapid release to activate intracellular calcium-dependent functions. Stromal interacting molecule 1 (STIM1) plays a critical role in early sensing of changes in the ER's calcium level, especially when there is a sudden release of stored calcium from the ER. Inactive STIM1, which has a bound calcium ion, is activated upon ion release. Following activation of STIM1, there is STIM1-assisted initiation of extracellular calcium entry through channels in the cell membrane. This extracellular calcium entering the cell then amplifies intracellular calcium-dependent actions. At the end of the process, ER levels of stored calcium are reestablished. The main focus of this work was to study the conformational changes accompanying homo- or heterodimerization of STIM1. For this purpose, the ER luminal portion of STIM1 (residues 58-236), which includes the sterile alpha motif (SAM) domain plus the calcium-binding EF-hand domains 1 and 2 attached to the STIM1 transmembrane region (TM), was modeled and embedded in a virtual membrane. Next, molecular dynamics simulations were performed to study the conformational changes that take place during STIM1 activation and subsequent protein-protein interactions. Indeed, the simulations revealed exposure of residues in the EF-hand domains, which may be important for dimerization steps. Altogether, understanding conformational changes in STIM1 can help in drug discovery when targeting this key protein in intracellular calcium functions.
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- 2017
37. Measuring Calumenin Impact on ER-Calcium Depletion Using Transient Calumenin Overexpression and Silencing
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Reginald Philippe, Olivier Mignen, Institut Necker Enfants-Malades (INEM - UM 111 (UMR 8253 / U1151)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Génétique moléculaire et génétique épidémiologique, Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM), Lymphocyte B et Auto-immunité (LBAI), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Brestois Santé Agro Matière (IBSAM), and Université de Brest (UBO)
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0301 basic medicine ,Artificial intelligence ,SERCA ,Bioinformatics ,[SDV]Life Sciences [q-bio] ,Western blot ,Calcium Measurement ,Transfection ,Calumenin ,03 medical and health sciences ,Machine learning ,Secretory pathway ,Calcium signaling ,Calcium metabolism ,030102 biochemistry & molecular biology ,Chemistry ,Endoplasmic reticulum ,ER calcium ,Cell biology ,Calcium ATPase ,030104 developmental biology ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,Rheumatologists ,Ca2+ measurement ,Microplate reader - Abstract
International audience; Calumenin is a secretory pathway protein regulating different endoplasmic reticulum (ER) proteins such as the sarco-endoplasmic reticulum calcium ATPase (SERCA) pumps. Combined with its diverse cellular distribution, its calcium-binding ability, and its interaction with proteins involved in calcium signaling, it is easy to speculate on future description of important roles of calumenin in calcium homeostasis in many cell types, as it was initially observed in muscle cells. In this chapter, we describe basic techniques to modulate calumenin expression and detect its impact on ER calcium content using classic transfection and Western blot techniques, as well as ER calcium measurement using microplate reader.
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- 2019
38. Overexpression of certain transient receptor potential and Orai channels in prostate cancer is associated with decreased risk of systemic recurrence after radical prostatectomy
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Laurent Doucet, Georges Fournier, Olivier Mignen, Pascale Marcorelles, S. Rosec, Arnaud Uguen, Antoine Valeri, N. Schoentgen, Marie Potier-Cartereau, Ronan Garlantézec, Gaëlle Fromont, Matthieu Talagas, Claude Férec, Marie-Aimée Perrouin-Verbe, Christophe Vandier, Génétique, génomique fonctionnelle et biotechnologies (UMR 1078) (GGB), Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO)-Université de Brest (UBO)-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre hospitalier universitaire de Nantes (CHU Nantes), Centre Hospitalier Régional Universitaire de Brest (CHRU Brest), Laboratoire sur les interactions Epithéliums Neurones (LIEN), Université de Brest (UBO), Institut de recherche en santé, environnement et travail (Irset), Université d'Angers (UA)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Centre d'Investigation Clinique (CIC - Brest), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM), Nutrition, croissance et cancer (U 1069) (N2C), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Tours, Centre Hospitalier Régional Universitaire de Tours (CHRU TOURS), Lymphocyte B et Auto-immunité (LBAI), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Brestois Santé Agro Matière (IBSAM), Ligue Contre le CancerLigue nationale contre le cancer, Association Francaise d'Urologie, EFS-Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Brestois Santé Agro Matière (IBSAM), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Santé et de la Recherche Médicale (INSERM)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Université d'Angers (UA), Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Hospitalier Régional Universitaire de Tours (CHRU Tours), Université de Brest (UBO)-Institut Brestois Santé Agro Matière (IBSAM), Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Lymphocytes B, Autoimmunité et Immunothérapies (LBAI), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-LabEX IGO Immunothérapie Grand Ouest, and Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ)-Institut Brestois Santé Agro Matière (IBSAM)
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0301 basic medicine ,Oncology ,Male ,Risk ,medicine.medical_specialty ,Surgical margin ,TRPV6 ,recurrence ,Urology ,[SDV]Life Sciences [q-bio] ,Lower risk ,calcium signaling ,03 medical and health sciences ,Prostate cancer ,0302 clinical medicine ,Transient Receptor Potential Channels ,Predictive Value of Tests ,Internal medicine ,Biomarkers, Tumor ,Medicine ,Humans ,ComputingMilieux_MISCELLANEOUS ,Aged ,Prostatectomy ,Tissue microarray ,business.industry ,Cancer ,Prostatic Neoplasms ,biomarkers ,Middle Aged ,medicine.disease ,prostate cancer ,Calcium Release Activated Calcium Channels ,radical prostatectomy ,3. Good health ,030104 developmental biology ,Real-time polymerase chain reaction ,030220 oncology & carcinogenesis ,Case-Control Studies ,Multivariate Analysis ,Immunohistochemistry ,Neoplasm Recurrence, Local ,business - Abstract
BACKGROUND Several studies had suggested the potential role of calcium signaling in prostate cancer (PCa) prognosis and agressiveness. We aimed to investigate selected proteins contributing to calcium (Ca2+ ) signaling, (Orai, stromal interaction molecule (STIM), and transient receptor potential (TRP) channels) and involved in cancer hallmarks, as independent predictors of systemic recurrence after radical prostatectomy (RP). METHODS A case-control study including 112 patients with clinically localized PCa treated by RP between 2002 and 2009 and with at least 6-years' follow-up. Patients were divided into two groups according to the absence or presence of systemic recurrence. Expression levels of 10 proteins involved in Ca2+ signaling (TRPC1, TRPC4, TRPV5, TRPV6, TRPM8, STIM1, STIM2, Orai1, Orai2, and Orai3), were assessed by immunohistochemistry using tissue microarrays (TMAs) constructed from paraffin-embedded PCa specimens. The level of expression of the various transcripts in PCa was assessed using quantitative polymerase chain reaction (qPCR) analysis. RNA samples for qPCR were obtained from fresh frozen tissue samples of PCa after laser capture microdissection on RP specimens. Relative gene expression was analyzed using the 2-▵▵Ct method. RESULTS Multivariate analysis showed that increased expression of TRPC1, TRPC4, TRPV5, TRPV6, TRPM8, and Orai2 was significantly associated with a lower risk of systemic recurrence after RP, independently of the prostate-specific antigen (PSA) level, percentage of positive biopsies, and surgical margin (SM) status (P = .007, P = .01, P
- Published
- 2019
39. A Practical Access to New Pyrazole SKF-96365 Analogues as Potential Store-Operated Calcium Entry (SOCE) Inhibitors
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Janat Mamybékova, Olivier Mignen, Yves-Alain Bekro, Camille Déliko Dago, Bazureau Jean Pierre, Christophe Brigaudeau, Thierry Roisnel, and Lou-Anna Voli
- Subjects
chemistry.chemical_compound ,Chemistry ,Hydrochloride ,Endoplasmic reticulum ,Diastereomer ,Substituent ,Side chain ,Enantiomer ,Pyrazole ,Store-operated calcium entry ,Medicinal chemistry - Abstract
The racemic synthesis in four steps of pyrazole SKF-96365 analogues without substituent (CF3 group) on the pyrazole platform was realized in moderate to good yields. The separation of (±) hydroxyl enantiomers 4 was developed successfully using the method of "half-concentration" with commercial (+)-(1S)- and (-)-(1R)-10-camphorsulfonic acid (CSA) followed by neutralization of diastereomers with MeONa in dry MeOH solution. With the pure enantiomers (-)-(1S)-4b and (+)-(1R)-4b, initial attempts to obtain the crystallized (-)-(1S)-7d and (+)-(1R)-7d after treatment of intermediate 6d with a solution of 1M HCl (for precipitation of hydrochloride salt 7d) failed. We have also investigated the effects of compounds 7(a-d) on endoplasmic reticulum (ER) Ca2+ and SOCE on PLP-B lymphocyte cell line and compound 7d was identified as a better SOCE inhibitor than SKF-96365. This preliminary SAR study showed that the MeO group in para-position of the phenethyl-1H-pyrazolium skeleton or for the Cbeta-phenylpropoxy side chain of 7 influenced the SOCE activity.
- Published
- 2018
40. La ciguatoxine-2 du Pacifique et la brévétoxine-1 entraînent la sensibilisation de récepteurs sensoriels impliqués dans le prurit
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Laurent Misery, Ophélie Pierre, Olivier Mignen, N. Le Goux, Paul Buscaglia, Raphael Leschiera, Maxime Fouchard, and R. Le Garrec
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Dermatology - Abstract
Introduction Les ciguatoxines (CTXs) et les brevetoxines (PbTxs) sont des neurotoxines impliquees respectivement dans la ciguatera et l’intoxication neurologique par les fruits de mer, deux syndromes consecutifs a l’ingestion de poissons ou de mollusques contamines. Les symptomes communs mais d’intensite plus marquee dans la ciguatera, sont des troubles neuro-cutanes (prurit, paresthesies et dysesthesies au froid) qui peuvent persister plusieurs semaines a plusieurs annees. Les cibles primaires de ces toxines sont le canal sodium dependant du potentiel de membrane (Nav) dont elles entrainent l’ouverture. Il s’ensuit une hyperexcitabilite membranaire des neurones peripheriques. Cependant, les evenements moleculaires en aval restent peu connus et aucun traitement n’existe pour soulager ces troubles neuro-cutanes persistants et invalidants. Recemment, nous avons mis en evidence que la Pacific-ciguatoxine-2 (P-CTX-2), appliquee a une co-culture de keratinocytes/neurones sensoriels, induit la liberation des neuropeptides de l’inflammation neurogene (CGRP et substance P). Chez la souris, il a ete montre que l’allodynie au froid induite par les CTXs implique la sensibilisation (i.e. l’abaissement du seuil d’activation) Nav-dependante de TRPA1. Nos travaux ont pour but d’etudier la sensibilisation de plusieurs recepteurs sensoriels cutanes par ces neurotoxines. Materiel et methodes Des cultures de neurones sensoriels de rats nouveaux-nes ont ete incubes pendant 24 h avec la P-CTX-2 ou la brevetoxine-1 (PbTx-1). A l’aide de la video-microscopie en fluorescence sur cellule unique, nous avons etudie la modulation par ces toxines de la reponse calcique a un agoniste specifique du recepteur sensoriel d’interet. Dans un second temps, nous avons dose dans le surnageant de co-cultures de keratinocytes et neurones sensoriels traitee par la P-CTX-2 plusieurs mediateurs susceptibles de contribuer a la sensibilisation de recepteurs sensoriels. Resultats Nos resultats montrent que la P-CTX-2 et la PbTx-1 sensibilisent differents recepteurs sensoriels de la peau. De plus, ils mettent en evidence que la P-CTX-2 augmente, dans la coculture la liberation de plusieurs mediateurs, suggerant leur role dans la sensibilisation des recepteurs sensoriels cutanes induite par les neurotoxines. Discussion Ce travail permet de mieux comprendre la physiopathologie des troubles sensoriels induits par les CTXs et les PbTxs et le prurit de maniere generale. Cette etude suggere que le mecanisme de sensibilisation mis en evidence pourrait participer a l’initiation et la persistance des troubles neuro-cutanees induits par la P-CTX-2 et la PbTx-1. Nos resultats et la caracterisation des mecanismes impliques dans la sensibilisation pourraient permettre d’identifier des voies therapeutiques ciblees. Ce travail a ete realise avec le soutien de la Societe francaise de dermatologie.
- Published
- 2020
41. Function and regulation of TRPM7, as well as intracellular magnesium content, are altered in cells expressing ΔF508-CFTR and G551D-CFTR
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Mathieu Kerbiriou, Pascal Trouvé, Marie-Laure Calvez, F. D. Horgen, Paul Buscaglia, S. Le Hir, Florentin Huguet, Claude Férec, Nathalie Benz, and Olivier Mignen
- Subjects
0301 basic medicine ,congenital, hereditary, and neonatal diseases and abnormalities ,Patch-Clamp Techniques ,Cystic Fibrosis ,Mutant ,Cystic Fibrosis Transmembrane Conductance Regulator ,TRPM Cation Channels ,Protein Serine-Threonine Kinases ,Biology ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,Adenosine Triphosphate ,Chloride Channels ,TRPM7 ,medicine ,Humans ,Magnesium ,Protein Interaction Maps ,RNA, Messenger ,Molecular Biology ,Magnesium ion ,Pharmacology ,Activator (genetics) ,Wild type ,Cell Biology ,respiratory system ,Molecular biology ,Naltrexone ,digestive system diseases ,Cystic fibrosis transmembrane conductance regulator ,respiratory tract diseases ,Kinetics ,030104 developmental biology ,Gene Expression Regulation ,Naltriben ,Monoterpenes ,Mutagenesis, Site-Directed ,Chloride channel ,biology.protein ,Cymenes ,Molecular Medicine ,Calcium ,Fura-2 ,HeLa Cells ,medicine.drug - Abstract
Cystic fibrosis (CF), one of the most common fatal hereditary disorders, is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. The CFTR gene product is a multidomain adenosine triphosphate-binding cassette (ABC) protein that functions as a chloride (Cl(-)) channel that is regulated by intracellular magnesium [Mg(2+)]i. The most common mutations in CFTR are a deletion of a phenylalanine residue at position 508 (ΔF508-CFTR, 70-80 % of CF phenotypes) and a Gly551Asp substitution (G551D-CFTR, 4-5 % of alleles), which lead to decreased or almost abolished Cl(-) channel function, respectively. Magnesium ions have to be finely regulated within cells for optimal expression and function of CFTR. Therefore, the melastatin-like transient receptor potential cation channel, subfamily M, member 7 (TRPM7), which is responsible for Mg(2+) entry, was studies and [Mg(2+)]i measured in cells stably expressing wildtype CFTR, and two mutant proteins (ΔF508-CFTR and G551D-CFTR). This study shows for the first time that [Mg(2+)]i is decreased in cells expressing ΔF508-CFTR and G551D-CFTR mutated proteins. It was also observed that the expression of the TRPM7 protein is increased; however, membrane localization was altered for both ΔF508del-CFTR and G551D-CFTR. Furthermore, both the function and regulation of the TRPM7 channel regarding Mg(2+) is decreased in the cells expressing the mutated CFTR. Ca(2+) influx via TRPM7 were also modified in cells expressing a mutated CFTR. Therefore, there appears to be a direct involvement of TRPM7 in CF physiopathology. Finally, we propose that the TRPM7 activator Naltriben is a new potentiator for G551D-CFTR as the function of this mutant increases upon activation of TRPM7 by Naltriben.
- Published
- 2016
42. Ciguatera poisoning: the role of high-voltage-activated and store-operated calcium channels in ciguatoxin-induced sensory effects
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Matthieu Talagas, Laurent Misery, Christelle Le Gall-Ianotto, Raphaele Le Garrec, Reginald Philippe, Killian L’Herondelle, Richard J. Lewis, and Olivier Mignen
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Ciguatoxin ,Voltage-dependent calcium channel ,Chemistry ,Biophysics ,Sensory system ,Ciguatera Poisoning - Published
- 2020
43. Preliminary Structure-Activity Relationship (SAR) of a Novel Series of Pyrazole SKF-96365 Analogues as Potential Store-Operated Calcium Entry (SOCE) Inhibitors
- Author
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Paul Le Maux, Olivier Mignen, Christophe Brigaudeau, Jean Pierre Bazureau, Camille Déliko Dago, Yves-Alain Bekro, Thierry Roisnel, Institut des Sciences Chimiques de Rennes (ISCR), Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie Bioorganique et des Substances Naturelles (LCBOSN), Université Abobo-Adjamé, Lymphocytes B, Autoimmunité et Immunothérapies (LBAI), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-LabEX IGO Immunothérapie Grand Ouest, Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO), CalciScreen Platform, Université Nangui Abrogoua (UNA), Synthèse Caractérisation Analyse de la Matière (ScanMAT), Université de Rennes (UR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Jonchère, Laurent, Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA), Lymphocyte B et Auto-immunité (LBAI), Université de Brest (UBO)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Nangui Abrogoua, Centre National de la Recherche Scientifique (CNRS)-Université de Rennes 1 (UR1), and Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut de Chimie du CNRS (INC)
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0301 basic medicine ,Hydrochloride ,Quantitative Structure-Activity Relationship ,pyrazole ,SKF-96365 ,analogues ,resolution ,half-quantities ,SOCE ,B lymphocyte cell ,SOCE inhibitor ,Ca2+ signalling ,Pyrazole ,Catalysis ,Article ,Inorganic Chemistry ,lcsh:Chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,Cell Line, Tumor ,[CHIM] Chemical Sciences ,Structure–activity relationship ,Humans ,[CHIM]Chemical Sciences ,Calcium Signaling ,Physical and Theoretical Chemistry ,Molecular Biology ,IC50 ,lcsh:QH301-705.5 ,Spectroscopy ,Chemistry ,Endoplasmic reticulum ,Organic Chemistry ,Resolution (electron density) ,Imidazoles ,General Medicine ,Calcium Channel Blockers ,Store-operated calcium entry ,3. Good health ,Computer Science Applications ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,Cell culture ,Biophysics - Abstract
International audience; From a series of (1, 1)-1[β-(phenylalkoxy)-(phenetyl)]-1-pyrazolium hydrochloride as new analogues of SKF-96365, one has an interesting effect for endoplasmic reticulum (ER) Carelease and store-operated Caentry (SOCE) (IC25 μM) on the PLP-B lymphocyte cell line. A successful resolution of (±) 1-phenyl-2-(1-pyrazol-1-yl)ethan-1-ol has been developed by using the method of "half-concentration" in the presence of (+)-(1)- or (-)-(1)-CSA.
- Published
- 2018
44. A new tool to test active ingredient using lactic acid in vitro, a help to understand cellular mechanism involved in stinging test: An example using a bacterial polysaccharide (Fucogel ® )
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Killian L’Herondelle, Laurent Misery, Nicolas Lebonvallet, Mehdi Sakka, Thibaut Saguet, Olivier Gouin, Christelle Le Gall-Ianotto, Olivier Mignen, Paul Buscaglia, Raphael Leschiera, Jean-Luc Carré, Jean-Luc Philbe, Université de Brest (UBO), Centre Hospitalier Régional Universitaire de Brest (CHRU Brest), and Laboratoire sur les interactions Epithéliums Neurones (LIEN)
- Subjects
Adult ,Keratinocytes ,0301 basic medicine ,[SDV]Life Sciences [q-bio] ,Pain ,TRPV Cation Channels ,Nerve Tissue Proteins ,Dermatology ,Substance P ,Pharmacology ,Polysaccharide ,PC12 Cells ,Biochemistry ,Sensitive skin ,Young Adult ,030207 dermatology & venereal diseases ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,In vivo ,[SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB] ,Animals ,Humans ,Lactic Acid ,Molecular Biology ,ComputingMilieux_MISCELLANEOUS ,Aged ,Skin ,Active ingredient ,chemistry.chemical_classification ,Neurogenic inflammation ,Polysaccharides, Bacterial ,Bacterial polysaccharide ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,[SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology ,Hydrogen-Ion Concentration ,Middle Aged ,Coculture Techniques ,In vitro ,Rats ,3. Good health ,Lactic acid ,Acid Sensing Ion Channels ,030104 developmental biology ,chemistry ,Female ,Carrier Proteins ,[SDV.MHEP.DERM]Life Sciences [q-bio]/Human health and pathology/Dermatology - Abstract
The stinging test is an in vivo protocol that evaluates sensitive skin using lactic acid (LA). A soothing sensation of cosmetics or ingredients can be also appreciated through a decrease in stinging score. To predict the soothing sensation of a product before in vivo testing, we developed a model based on an LA test and substance P (SP) release using a co-culture of human keratinocytes and NGF-differentiated PC12 cells. A bacterial fucose-rich polysaccharide present in Fucogel® was evaluated as the soothing molecule in the in vivo stinging test and our in vitro model. Excluding toxic concentrations, the release of SP was significant from 0.2% of lactic acid for the PC12 cells and from 0.1% of lactic acid for the keratinocytes. When the pH was adjusted to approximately 7.4, LA did not provoke SP release. At these concentrations of LA, 0.1% of polysaccharide showed a significant decrease in SP release from the two cellular types and in co-cultures without modifying the pH of the medium. In vivo, a stinging test using the polysaccharide showed a 30% decrease in prickling intensity vs the placebo in 19 women between the ages of 21 and 69. Our in vitro model is ethically interesting and is adapted for cosmetic ingredients screening because it does not use animal experimentation and limits human volunteers. Moreover, Fucogel® reduced prickling sensation as revealed by the in vivo stinging test and inhibits the neurogenic inflammation as showed by our new in vitro stinging test based on SP release.
- Published
- 2018
45. Involvement of cathepsin S and protease-activated receptor-2 in ciguatoxin-induced substance P release: new promising targets to treat ciguatera pruritus
- Author
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PIERRE, Ophélie, primary, L'HERONDELLE, Killian, primary, Sophie, Fouyet, primary, Raphaël, Leschiera, primary, LE GALL-IANOTTO, Christelle, primary, Paul, Buscaglia, primary, Olivier, Mignen, primary, Misery, Laurent, primary, and Garrec Raphaele, Le, primary
- Published
- 2019
- Full Text
- View/download PDF
46. The SigmaR1 chaperone drives breast and colorectal cancer cell migration by tuning SK3-dependent Ca2+ homeostasis
- Author
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Y Fourbon, Gaëlle Fromont, Lucie Clarysse, R Schiappa, Aurélie Chantôme, Marie Potier-Cartereau, Raphael Rapetti-Mauss, Maxime Guéguinou, Pierre-Marie Martin, Christophe Vandier, Arnaud Uguen, E Chamorey, B Pellissier, D Crottès, Pascal Jézéquel, Franck Borgese, Olivier Mignen, Alban Girault, Olivier Soriani, Catherine Guérin-Charbonnel, Nutrition, croissance et cancer (U 1069) (N2C), Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Hospitalier Régional Universitaire de Tours (CHRU Tours), Laboratoire des technologies de la microélectronique (LTM ), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Dept. of Statistics, Centre de Lutte contre le Cancer Antoine Lacassagne [Nice] (UNICANCER/CAL), UNICANCER-Université Côte d'Azur (UCA)-UNICANCER-Université Côte d'Azur (UCA), Lymphocyte B et Auto-immunité (LBAI), Université de Brest (UBO)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Hospitalier Régional Universitaire de Brest (CHRU Brest), Institute of Developmental Biology and Cancer (IBDC), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), Lymphocytes B, Autoimmunité et Immunothérapies (LBAI), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-LabEX IGO Immunothérapie Grand Ouest, Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO), Université Nice Sophia Antipolis (1965 - 2019) (UNS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Tours, and Centre Hospitalier Régional Universitaire de Tours (CHRU TOURS)
- Subjects
0301 basic medicine ,Cancer Research ,medicine.medical_specialty ,Colorectal cancer ,[SDV]Life Sciences [q-bio] ,chemistry.chemical_element ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Calcium ,Biology ,Molecular oncology ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,Genetics ,medicine ,Molecular Biology ,ComputingMilieux_MISCELLANEOUS ,Calcium signaling ,Calcium metabolism ,ORAI1 ,Calcium channel ,Bone metastasis ,medicine.disease ,3. Good health ,030104 developmental biology ,Endocrinology ,chemistry ,030220 oncology & carcinogenesis ,Cancer research - Abstract
The remodeling of calcium homeostasis contributes to the cancer hallmarks and the molecular mechanisms involved in calcium channel regulation in tumors remain to be characterized. Here, we report that SigmaR1, a stress-activated chaperone, is required to increase calcium influx by triggering the coupling between SK3, a Ca2+-activated K+ channel (KCNN3) and the voltage-independent calcium channel Orai1. We show that SigmaR1 physically binds SK3 in BC cells. Inhibition of SigmaR1 activity, either by molecular silencing or by the use of sigma ligand (igmesine), decreased SK3 current and Ca2+ entry in breast cancer (BC) and colorectal cancer (CRC) cells. Interestingly, SigmaR1 inhibition diminished SK3 and/or Orai1 levels in lipid nanodomains isolated from BC cells. Analyses of tissue microarray from CRC patients showed higher SigmaR1 expression levels in cancer samples and a correlation with tumor grade. Moreover, the exploration of a cohort of 4937 BC patients indicated that high expression of SigmaR1 and Orai1 channels was significantly correlated to a lower overall survival. As the SK3/Orai1 tandem drives invasive process in CRC and bone metastasis progression in BC, our results may inaugurate innovative therapeutic approaches targeting SigmaR1 to control the remodeling of Ca2+ homeostasis in epithelial cancers.
- Published
- 2017
47. Calcium Signaling: From Normal B Cell Development to Tolerance Breakdown and Autoimmunity
- Author
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Patrice Hemon, Nelig Le Goux, Wesley H. Brooks, Marjolaine Debant, Yves Renaudineau, Sreya Mukherjee, Olivier Mignen, Michel, Geneviève, Lymphocyte B et Auto-immunité (LBAI), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO), European Space Research and Technology Centre (ESTEC), European Space Agency (ESA), Laboratoire d'Immunologie et Immunothérapie [Brest], Centre Hospitalier Régional Universitaire de Brest (CHRU Brest), School of Education Technology [Kolkata, West Bengal], Jadavpur University, Department of Chemistry, University of South Florida, Tampa, FL, USA., and University of South Florida [Tampa] (USF)
- Subjects
0301 basic medicine ,TRPC ,[SDV.IMM] Life Sciences [q-bio]/Immunology ,B-cell receptor ,Receptors, Antigen, B-Cell ,Context (language use) ,Autoimmunity ,Biology ,medicine.disease_cause ,Lymphocyte Activation ,Autoantigens ,Clonal deletion ,Autoimmune Diseases ,03 medical and health sciences ,Systemic lupus erythematosus ,0302 clinical medicine ,Primary Sjӧgren’s syndrome ,Calcium pathway ,medicine ,Immune Tolerance ,Immunology and Allergy ,Animals ,Humans ,Calcium Signaling ,Clonal Selection, Antigen-Mediated ,B cell ,Calcium signaling ,B cells ,B-Lymphocytes ,ORAI ,Receptor editing ,Cell Differentiation ,General Medicine ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Immunology ,STIM ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,Signal transduction ,Tolerance ,030215 immunology - Abstract
International audience; Maintenance of self-tolerance of auto-reactive lymphocytes is a fundamental mechanism to prevent the onset of autoimmune diseases. Deciphering the mechanisms involved in the deregulations leading to tolerance disruption and autoimmunity is still a major area of interest to identify new therapeutic targets and options. Ca(2+) signaling plays a major role in B cell normal development and is therefore finely tuned by B cell receptor (BCR)-dependent and independent pathways. Developmental changes in the characteristics of BCR-dependent Ca(2+) signals as well as the modulation of basal intracellular concentration ([Ca(2+)]i) contribute strongly to self-tolerance maintaining mechanisms responsible for the physical or functional elimination of autoreactive B cells such as clonal deletion, receptor editing, and anergy. Implication of Ca(2+) signals in B tolerance mechanisms mainly occurs through the specific activation of transcriptional programs depending on the amplitude, shape, and duration of Ca(2+) signals. A large number of studies reported Ca(2+) signaling defects in autoimmune pathology such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA) and primary Sjӧgren's syndrome (pSS). However, the precise nature of the molecular events responsible for these deregulations is not fully understood. Moreover, the demonstration of a direct correlation between Ca(2+) signaling defects and tolerance disruption is still lacking. The recent identification of proteins involved in B cell Ca(2+) signals such as ORAI, stromal interaction molecule and transient receptor potential is opening new horizons for understanding Ca(2+) signaling defects observed in autoimmune diseases and for proposing potentially new therapeutic solutions. This review aims to present an overview of the developmental evolution of BCR dependent Ca(2+) signaling and to place this signaling pathway in the context of mechanisms involved in tolerance maintenance and breakdown.
- Published
- 2017
48. Calumenin contributes to ER-Ca(2+) homeostasis in bronchial epithelial cells expressing WT and F508del mutated CFTR and to F508del-CFTR retention
- Author
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Maud Frieden, Florentin Huguet, Paul Buscaglia, Pascal Trouvé, Olivier Mignen, Fabrice Antigny, Caroline Norez, Cyril Castelbou, Frédéric Becq, Claude Férec, Reginald Philippe, Génétique, génomique fonctionnelle et biotechnologies (UMR 1078) (GGB), EFS-Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO), Hypertension arterielle pulmonaire physiopathologie et innovation thérapeutique, Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre chirurgical Marie Lannelongue, Lymphocyte B et Auto-immunité (LBAI), Université de Brest (UBO)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM), Signalisation et Transports Ioniques Membranaires (STIM), Université de Poitiers-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), Centre Hospitalier Universitaire de Purpan (CHU Purpan), Département de Physiologie Cellulaire et Métabolisme, Université de Genève (UNIGE), ONERA - The French Aerospace Lab [Palaiseau], ONERA-Université Paris Saclay (COmUE), Université de Bretagne Occidentale - UFR Médecine et Sciences de la Santé (UBO UFR MSS), Centre chirurgical Marie Lannelongue-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Brestois Santé Agro Matière (IBSAM), Université de Poitiers-Université de Tours-Centre National de la Recherche Scientifique (CNRS), ONERA, Institut Brestois Santé Agro Matière (IBSAM), and Université de Brest (UBO)-Université de Brest (UBO)-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)
- Subjects
0301 basic medicine ,medicine.medical_specialty ,congenital, hereditary, and neonatal diseases and abnormalities ,SERCA ,Physiology ,Biology ,medicine.disease_cause ,Calumenin ,Cystic fibrosis ,F508del-CFTR ,03 medical and health sciences ,0302 clinical medicine ,Calcium-binding protein ,Internal medicine ,medicine ,ddc:612 ,ER Ca(2+) homeostasis ,Molecular Biology ,Mutation ,Endoplasmic reticulum ,Wild type ,Cell Biology ,Transmembrane protein ,Cystic fibrosis transmembrane conductance regulator ,Cell biology ,030104 developmental biology ,Endocrinology ,Chloride channel ,biology.protein ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,030217 neurology & neurosurgery - Abstract
International audience; Cystic Fibrosis (CF) is the most frequent fatal genetic disease in Caucasian populations. Mutations in the chloride channel CF Transmembrane Conductance Regulator (CFTR) gene are responsible for functional defects of the protein and multiple associated dysregulations. The most common mutation in patients with CF, F508del-CFTR, causes defective CFTR protein folding. Thus minimal levels of the receptor are expressed at the cell surface as the mutated CFTR is retained in the endoplasmic reticulum (ER) where it correlates with defective calcium (Ca(2+)) homeostasis. In this study, we discovered that the Ca(2+) binding protein Calumenin (CALU) is a key regulator in the maintenance of ER-Ca(2+) calcium homeostasis in both wild type and F508del-CFTR expressing cells. Calumenin modulates SERCA pump activity without drastically affecting ER-Ca(2+) concentration. In addition, reducing Calumenin expression in CF cells results in a partial restoration of CFTR activity, highlighting a potential function of Calumenin in CFTR maturation. These findings demonstrate a pivotal role for Calumenin in CF cells, providing insights into how modulation of Calumenin expression or activity may be used as a potential therapeutic tool to correct defects in F508del-CFTR.
- Published
- 2017
49. Calumenin contributes to ER-Ca
- Author
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Réginald, Philippe, Fabrice, Antigny, Paul, Buscaglia, Caroline, Norez, Florentin, Huguet, Cyril, Castelbou, Pascal, Trouvé, Frédéric, Becq, Maud, Frieden, Claude, Férec, and Olivier, Mignen
- Subjects
Calcium-Binding Proteins ,Cystic Fibrosis Transmembrane Conductance Regulator ,Homeostasis ,Humans ,Bronchi ,Calcium ,Epithelial Cells ,Endoplasmic Reticulum ,Cells, Cultured - Abstract
Cystic Fibrosis (CF) is the most frequent fatal genetic disease in Caucasian populations. Mutations in the chloride channel CF Transmembrane Conductance Regulator (CFTR) gene are responsible for functional defects of the protein and multiple associated dysregulations. The most common mutation in patients with CF, F508del-CFTR, causes defective CFTR protein folding. Thus minimal levels of the receptor are expressed at the cell surface as the mutated CFTR is retained in the endoplasmic reticulum (ER) where it correlates with defective calcium (Ca
- Published
- 2017
50. Differentiation of human skin-derived precursors (SKPs) into functional sensory neurons
- Author
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Olivier, Mignen
- Abstract
Skin-derived precursor cells (SKPs) have been shown as a population of stem cells that express some markers common to the neural crest stem cells (NCSCs) as progenitor neural nestin, the low affinity neurotrophin receptor p75 and transcription factors as Sox9 and 10 and Pax3, Slug, Snail. SKPs are NCSCs that persist in certain adult tissues, particularly in the skin, which can generate a large part of the peripheral nervous system, particularly sensory neurons (SN). In this study, we reported a protocol to obtain SNs from SKPs. For this, we rely on a protocol already established by Reinhardt et al. in 2013 from Embryonic stem cells (ESCs) and induced Pluripotent stem cells (iPSCs). The SKPs were induced to generate SNs by treating cells with trophic factors and / or chemicals that had previously been shown to be important for the generation of SNs: Purmorphamine, CHIR99021, BMP4 and GDNF, BDNF, NGF. We show that the differentiation of SKPs into SNs was regulated by neurogenins 1, 2 and 3. At the end of the differentiation, the protocol allowed gene transcription of BRN3A and PRDM12 (marker of pain-sensing nerve cells). Expression was from 1000 to 2500 times higher for PRDM12 and BRN3A respectively versus undifferentiated SKP. Using immunostaining we showed that 65% and 80% of cells expressed BRN3A and peripherin, marker for peripheral neurons. Furthermore, cells expressed TRPV1, PAR2, TRPA1, substance P, CGRP, H1, B3-tubulin. Using calcium imaging, a proportion of cells was responding for histamine, SLIGKV (a specific agonist of PAR2), polygodial (a specific agonist of TRPA1), and capsaicin (a specific agonist of TRPV1). In conclusion, SKP are NCSC, that able to differentiate directly into functional sensory neuron.
- Published
- 2017
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