A new approach for studying GPCR dimers: Drug-induced inactivation and reactivation to reveal GPCR dimer function in vitro, in primary culture, and in vivo

Abstract Imidazolidinyl urea (IU) is used as an antimicrobial preservative in cosmetic and pharmaceutical products. IU induces allergic contact dermatitis, however, the mechanism has not yet been elucidated. Mas-related G protein-coupled receptor-X2 (MRGPRX2) triggers drug-induced pseudo-allergic reactions. The aims of this study were to determine whether IU activated mast cells through MRGPRX2 to further trigger contact dermatitis. Wild-type (WT) and KitW-sh/HNihrJaeBsmJNju (MUT) mice were treated with IU to observe its effects on local inflammation and mast cells degranulation in vivo. Laboratory of allergic disease 2 cells were used to detect calcium mobilization and release of inflammatory mediators in vitro. WT mice showed a severe local inflammatory response and contact dermatitis, whereas only slight inflammatory infiltration was observed in MUT mice. Thus, MRGPRX2 mediated the IU-induced activation of mast cells. However, histamine, a typical allergen, was not involved in this process. Tryptase expressed by mast cells was the major non-histaminergic inflammatory mediator of contact dermatitis. IU induced anaphylactic reaction via MRGPRX2 and further triggering non-histaminergic contact dermatitis, which explained why antihistamines are clinically ineffective against some chronic dermatitis.

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In Vivo and In Vitro Toxicodynamic Analyses of New Quinolone-and Nonsteroidal Anti-Inflammatory Drug-Induced Effects on the Central Nervous System

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.43.5.1091 ◽

1999 ◽

Vol 43 (5) ◽

pp. 1091-1097 ◽

Cited By ~ 7

Author(s):

Hideki Kita ◽

Hirotami Matsuo ◽

Hitomi Takanaga ◽

Junichi Kawakami ◽

Koujirou Yamamoto ◽

...

Keyword(s):

Brain Tissue ◽

Threshold Concentration ◽

Current Response ◽

Drug Induced ◽

Inhibition Ratio ◽

New Quinolones ◽

The Central Nervous System ◽

Anti Inflammatory

ABSTRACT We investigated the correlation between an in vivo isobologram based on the concentrations of new quinolones (NQs) in brain tissue and the administration of nonsteroidal anti-inflammatory drugs (NSAIDs) for the occurrence of convulsions in mice and an in vitro isobologram based on the concentrations of both drugs for changes in the γ-aminobutyric acid (GABA)-induced current response in Xenopus oocytes injected with mRNA from mouse brains in the presence of NQs and/or NSAIDs. After the administration of enoxacin (ENX) in the presence or absence of felbinac (FLB), ketoprofen (KTP), or flurbiprofen (FRP), a synergistic effect was observed in the isobologram based on the threshold concentration in brain tissue between mice with convulsions and those without convulsions. The three NSAIDs did not affect the pharmacokinetic behavior of ENX in the brain. However, the ENX-induced inhibition of the GABA response in the GABAA receptor expressed in Xenopus oocytes was enhanced in the presence of the three NSAIDs. The inhibition ratio profiles of the GABA responses for both drugs were analyzed with a newly developed toxicodynamic model. The inhibitory profiles for ENX in the presence of NSAIDs followed the order KTP (1.2 μM) > FRP (0.3 μM) > FLB (0.2 μM). These were 50- to 280-fold smaller than those observed in the absence of NSAIDs. The inhibition ratio (0.01 to 0.02) of the GABAA receptor in the presence of both drugs was well-fitted to the isobologram based on threshold concentrations of both drugs in brain tissue between mice with convulsions and those without convulsions, despite the presence of NSAIDs. In mice with convulsions, the inhibitory profiles of the threshold concentrations of both drugs in brain tissue of mice with convulsions and those without convulsions can be predicted quantitatively by using in vitro GABA response data and toxicodynamic model.

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Drug-induced liver injury classification model based on in vitro human transcriptomics and in vivo rat clinical chemistry data

Systems Biomedicine ◽

10.4161/sysb.29400 ◽

2014 ◽

Vol 2 (4) ◽

pp. 63-70 ◽

Cited By ~ 10

Author(s):

Danyel Jennen ◽

Jan Polman ◽

Mark Bessem ◽

Maarten Coonen ◽

Joost van Delft ◽

...

Keyword(s):

Liver Injury ◽

Clinical Chemistry ◽

Classification Model ◽

Drug Induced ◽

Drug Induced Liver Injury ◽

Chemistry Data ◽

Model Based ◽

Injury Classification

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Bioresorbable Microspheres as Injectable Cell Carrier: FRom Preparation to in Vitro Evaluation

MRS Proceedings ◽

10.1557/proc-550-183 ◽

1998 ◽

Vol 550 ◽

Cited By ~ 1

Author(s):

Y. Senuma ◽

S. Franceschin ◽

J. G. Hilborn ◽

P. Tissiéres ◽

P. Frey

Keyword(s):

Cultured Cells ◽

Urothelial Cells ◽

New Approach ◽

Muscular Structure ◽

Cell Carrier ◽

Support Matrix ◽

Vesico Ureteral Reflux

AbstractA new approach to the vesico-ureteral reflux could be a local regeneration of the defective vesicoureteral junction by transplanting living cells to the target site. The aim of this work is to provide a long-term effective treatment by producing bioresorbable microspheres which can act as support matrix for those cells, with the goal of an in vivo transfer of the in vitro cultured cells with a minimal surgical procedure. After microsphere degradation, the cells should be integrated into the muscular structure of the junction. Most innovative is that these are cultured muscle and urothelial cells from the bladder of the same patient.

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A New Approach to Studying Ochratoxin A (OTA)-Induced Nephrotoxicity: Expression Profiling in Vivo and in Vitro Employing cDNA Microarrays

Toxicological Sciences ◽

10.1093/toxsci/kfg073 ◽

2003 ◽

Vol 73 (2) ◽

pp. 315-328 ◽

Cited By ~ 110

Author(s):

A. Luhe

Keyword(s):

Ochratoxin A ◽

Expression Profiling ◽

Cdna Microarrays ◽

New Approach

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A Tissue-Engineered Tracheobronchial In Vitro Co-Culture Model for Determining Epithelial Toxicological and Inflammatory Responses

Biomedicines ◽

10.3390/biomedicines9060631 ◽

2021 ◽

Vol 9 (6) ◽

pp. 631

Author(s):

Luis Soriano ◽

Tehreem Khalid ◽

Fergal J. O'Brien ◽

Cian O'Leary ◽

Sally-Ann Cryan

Keyword(s):

Epithelial Cells ◽

Inflammatory Responses ◽

Bronchial Epithelial Cells ◽

Lung Fibroblasts ◽

Drug Induced ◽

Bronchial Epithelial ◽

Culture Model ◽

Epithelial Cultures

Translation of novel inhalable therapies for respiratory diseases is hampered due to the lack of in vitro cell models that reflect the complexity of native tissue, resulting in many novel drugs and formulations failing to progress beyond preclinical assessments. The development of physiologically-representative tracheobronchial tissue analogues has the potential to improve the translation of new treatments by more accurately reflecting in vivo respiratory pharmacological and toxicological responses. Herein, advanced tissue-engineered collagen hyaluronic acid bilayered scaffolds (CHyA-B) previously developed within our group were used to evaluate bacterial and drug-induced toxicity and inflammation for the first time. Calu-3 bronchial epithelial cells and Wi38 lung fibroblasts were grown on either CHyA-B scaffolds (3D) or Transwell® inserts (2D) under air liquid interface (ALI) conditions. Toxicological and inflammatory responses from epithelial monocultures and co-cultures grown in 2D or 3D were compared, using lipopolysaccharide (LPS) and bleomycin challenges to induce bacterial and drug responses in vitro. The 3D in vitro model exhibited significant epithelial barrier formation that was maintained upon introduction of co-culture conditions. Barrier integrity showed differential recovery in CHyA-B and Transwell® epithelial cultures. Basolateral secretion of pro-inflammatory cytokines to bacterial challenge was found to be higher from cells grown in 3D compared to 2D. In addition, higher cytotoxicity and increased basolateral levels of cytokines were detected when epithelial cultures grown in 3D were challenged with bleomycin. CHyA-B scaffolds support the growth and differentiation of bronchial epithelial cells in a 3D co-culture model with different transepithelial resistance in comparison to the same co-cultures grown on Transwell® inserts. Epithelial cultures in an extracellular matrix like environment show distinct responses in cytokine release and metabolic activity compared to 2D polarised models, which better mimic in vivo response to toxic and inflammatory stimuli offering an innovative in vitro platform for respiratory drug development.

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In vivo measurement of intrauterine pressure by telemetry: a new approach for studying parturition in mouse models

Physiological Genomics ◽

10.1152/physiolgenomics.00058.2010 ◽

2010 ◽

Vol 42 (2) ◽

pp. 310-316 ◽

Cited By ~ 12

Author(s):

Stephanie L. Pierce ◽

William Kutschke ◽

Rafael Cabeza ◽

Sarah K. England

Keyword(s):

Mouse Models ◽

Accurate Method ◽

Isometric Tension ◽

Mouse Uterus ◽

New Approach ◽

In Vivo Measurement ◽

Intrauterine Pressure

Transgenic and knockout mouse models have proven useful in the study of genes necessary for parturition—including genes that affect the timing and/or progression of labor contractions. However, taking full advantage of these models will require a detailed characterization of the contractile patterns in the mouse uterus. Currently the best methodology for this has been measurement of isometric tension in isolated muscle strips in vitro. However, this methodology does not provide a real-time measure of changes in uterine pressure over the course of pregnancy. Recent advances have opened the possibility of using radiotelemetric devices to more accurately and comprehensively study intrauterine pressure in vivo. We tested the effectiveness of this technology in the mouse, in both wild-type (WT) mice and a mouse model of defective parturition (SK3 channel-overexpressing mice), after surgical implant of telemetry transmitters into the uterine horn. Continuous recordings from day 18 of pregnancy through delivery revealed that WT mice typically deliver during the 12-h dark cycle after 19.5 days postcoitum. In these mice, intrauterine pressure gradually increases during this cycle, to threefold greater than that measured during the 12-h cycle before delivery. SK3-overexpressing mice, by contrast, exhibited lower intrauterine pressure over the same period. These results are consistent with the outcome of previous in vitro studies, and they indicate that telemetry is an accurate method for measuring uterine contraction, and hence parturition, in mice. The use of this technology will lead to important novel insights into changes in intrauterine pressure during the course of pregnancy.

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Remodeling tumor microenvironment by liposomal co-delivery of DMXAA and simvastatin inhibits malignant melanoma progression

10.1101/2021.04.26.441259 ◽

2021 ◽

Author(s):

Valentin Florian Rauca ◽

Laura Patras ◽

Lavinia Luput ◽

Emilia Licarete ◽

Vlad Alexandru Toma ◽

...

Keyword(s):

Tumor Microenvironment ◽

Hypoxia Inducible Factor ◽

Tumor Development ◽

Novel Therapy ◽

Drug Induced ◽

Angiogenic Therapy ◽

Melanoma Model ◽

M1 Phenotype

Anti-angiogenic therapies for melanoma have not yet been translated into meaningful clinical benefit for patients, due to development of drug-induced resistance in cancer cells, mainly caused by hypoxia-inducible factor 1α (HIF-1α) overexpression and enhanced oxidative stress mediated by tumor-associated macrophages (TAMs). Our previous study demonstrated synergistic antitumor actions of simvastatin (SIM) and 5,6-dimethylxanthenone-4-acetic acid (DMXAA) on an in vitro melanoma model via suppression of the aggressive phenotype of melanoma cells and inhibition of TAMs-mediated angiogenesis. Therefore, we took the advantage of long circulating liposomes (LCL) superior tumor targeting capacity to efficiently deliver SIM and DMXAA to B16.F10 melanoma in vivo, with the final aim of improving the outcome of the anti-angiogenic therapy. Thus, we assessed the effects of this novel combined tumor-targeted treatment on s.c. B16.F10 murine melanoma growth and on the production of critical markers involved in tumor development and progression. Our results showed that the combined liposomal therapy inhibited almost totally the growth of melanoma tumors, due to the enhancement of anti-angiogenic effects of LCL-DMXAA by LCL-SIM and induction of a pro-apoptotic state in the tumor microenvironment (TME). These effects were favoured by the partial re-education of TAMs towards a M1 phenotype and maintained via suppression of major invasion and metastasis promoters (HIF-1α, pAP-1 c-Jun, and MMPs). Thus, this novel therapy holds the potential to remodel the tumor microenvironment, by suppressing its most important malignant biological capabilities.

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IN VITRO AND IN VIVO EVALUATION OF 2-CHLOROE THYLN ITROSOUREA DERIVATIVES AS ANTITUMOR AGENTS

Experimental Oncology ◽

10.31768/2312-8852.2015.37(1):23-29 ◽

2015 ◽

Vol 37 (1) ◽

pp. 23-29

Author(s):

A Sen ◽

K K Goswami ◽

A Mallick ◽

A K Saxena ◽

U Sanyal ◽

...

Keyword(s):

T Cells ◽

Antitumor Agents ◽

Malignant Tumors ◽

Optimum Dose ◽

Mouse Tumor ◽

Drug Induced ◽

In Vivo Evaluation ◽

Liver And Kidney

Aim: To evaluate potential of Naphthal-NU, Napro-NU and 5-Nitro-naphthal-NU, 2-chloroethylnitrosourea compounds with substituted naphthalimide in the pre-clinical studies. Materials and Methods: In vitro cytotoxicity of three nitrosoureas was determined in human and mouse tumor cell lines by MTT assays. In vivo anti-tumor potential was evaluated in Sarcoma-180 (S-180) and Ehrlich’s carcinoma (EC) solid tumors. Apoptosis in S-180 cells was analyzed by using Annexin V-Propidium Iodide (PI). Histological analysis of liver and kidney was performed at optimum dose (50 mg/kg). Expression status of CD4+, CD8+ and CD25+ cells in treated mouse were also examined. Results: Significant tumor growth retardation by the compounds was noted in early and advanced disease groups, as the life span of drug treated mice increased considerably. Drug induced killing was observed by induction of apoptosis. Naphthal-NU and 5-Nitro-naphthal-NU were effective to normalize the tumor induced structural abnormalities of liver and kidney. The compounds have no immunotoxic effect on CD4+ and CD8+ T cells and down regulate CD4+CD25+ regulatory T cells. Conclusion: Overall data holds promise for the antitumor activity with lower toxicity of the compounds that can be utilized for the treatment of human malignant tumors.

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