Isolation and characterization of a mouse L cell variant deficient in glucocorticoid receptors

Housley, Paul R.; Forsthoefel, Antonia M.

doi:10.1016/0006-291x(89)91745-2

Cited by 34 publications

(16 citation statements)

References 29 publications

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“…E8.2 cells, derived from mouse L929 fibroblasts (Housley and Forshoefel, 1989), are spontaneous glucocorticoid-resistant cells and contain neither detectable GR protein nor mRNA transcripts, whereas in E8.2/GR3 cells the GR protein levels are reconstituted and are regulated by tetracycline both temporally and in a dose-dependent manner as shown previously (Wei et al, 1998).…”

Section: Methodsmentioning

confidence: 55%

Mechanism of the Anti-Inflammatory Effect of Thiazolidinediones: Relationship with the Glucocorticoid Pathway

Ialenti¹,

Grassia²,

Meglio³

et al. 2005

Mol Pharmacol

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The glucocorticoid receptor (GR) and peroxisome proliferatoractivated receptors (PPARs) play important roles in both physiological and pathological conditions such as cell differentiation, lipolysis, control of glucose metabolism, immunity, and inflammation. In fact, recent studies suggest that the thiazolidinedione (TZD) class of PPAR-␥ ligands, like glucocorticoids, may also be clinically beneficial in several inflammatory diseases, even if the molecular mechanisms responsible for these activities have not yet been clarified. In this study, by using a murine model of inflammation, the carrageenin-induced paw edema in mouse, we show that the anti-inflammatory activity exhibited by the PPAR-␥ agonists rosiglitazone and ciglitazone is reversed by the GR antagonist RU486 (17␤-hydroxy-11␤-[4-dimethylamino phenyl]-17␣-[1-propynyl]estra-4,9-dien-3-one). Moreover, by using a conditional GR null cell line, we demonstrate, for the first time to our knowledge, that one of the possible mechanisms explaining the anti-inflammatory activity of TZDs is their ability to activate GR nuclear translocation. In addition, by using J774 cell line lacking PPAR-␥, we demonstrate that PPAR-␥ expression could not be essential for TZDmediated GR nuclear translocation, thus explaining, at least in part, the molecular mechanism underlying their anti-inflammatory activity.

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Section: Methodsmentioning

confidence: 55%

Mechanism of the Anti-Inflammatory Effect of Thiazolidinediones: Relationship with the Glucocorticoid Pathway

Ialenti¹,

Grassia²,

Meglio³

et al. 2005

Mol Pharmacol

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“…First, we tested whether the lack of GR in GR null mutant fibroblasts affects the ability of drugs to inhibit inflammatory transcripts, which are predominantly controlled by NF‐κB. Absence of the GR in this spontaneous mutant line was previously selected for (Housley & Forsthoefel, ) and confirmed here through Western blot (Fig D). Cells were then treated with prednisolone or VBP15 and inflammatory transcripts were induced with TNFα.…”

Section: Resultsmentioning

confidence: 84%

VBP15, a novel anti‐inflammatory and membrane‐stabilizer, improves muscular dystrophy without side effects

et al. 2013

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Absence of dystrophin makes skeletal muscle more susceptible to injury, resulting in breaches of the plasma membrane and chronic inflammation in Duchenne muscular dystrophy (DMD). Current management by glucocorticoids has unclear molecular benefits and harsh side effects. It is uncertain whether therapies that avoid hormonal stunting of growth and development, and/or immunosuppression, would be more or less beneficial. Here, we discover an oral drug with mechanisms that provide efficacy through anti-inflammatory signaling and membrane-stabilizing pathways, independent of hormonal or immunosuppressive effects. We find VBP15 protects and promotes efficient repair of skeletal muscle cells upon laser injury, in opposition to prednisolone. Potent inhibition of NF-κB is mediated through protein interactions of the glucocorticoid receptor, however VBP15 shows significantly reduced hormonal receptor transcriptional activity. The translation of these drug mechanisms into DMD model mice improves muscle strength, live-imaging and pathology through both preventive and post-onset intervention regimens. These data demonstrate successful improvement of dystrophy independent of hormonal, growth, or immunosuppressive effects, indicating VBP15 merits clinical investigation for DMD and would benefit other chronic inflammatory diseases.

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“…To determine whether this was the case, GR immune pellets were incubated with concentrated cytosol from E82.A3 cells. The E82 subline of L929 fibroblasts was selected for glucocorticoid resistance, and E82 cells do not contain any GR mRNA or GR protein (29). As shown in Fig.…”

Section: Fig 2 Dynamitin Is Present In Gr Heterocomplexes a Immunmentioning

confidence: 99%

Evidence for Glucocorticoid Receptor Transport on Microtubules by Dynein

Harrell

Murphy

Morishima

et al. 2004

Journal of Biological Chemistry

114

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Rapid, ligand-dependent movement of glucocorticoid receptors (GR) from cytoplasm to the nucleus is hsp90-dependent, and much of the movement system has been defined. GR⅐hsp90 heterocomplexes isolated from cells contain one of several hsp90-binding immunophilins that link the complex to cytoplasmic dynein, a molecular motor that processes along microtubular tracks to the nucleus. Although it is known that rapid, hsp90-dependent GR movement requires intact microtubules, it has not been shown that the movement is dynein-dependent. Here, we show that overexpression of dynamitin, which blocks movement by dissociating the dynein motor from its cargo, inhibits ligand-dependent movement of the GR to the nucleus. We show that native GR⅐hsp90⅐immnunophilin complexes contain dynamitin as well as dynein and that GR heterocomplexes isolated from cytosol containing paclitaxel and GTP to stabilize microtubules also contain tubulin. The complete movement system, including the dynein motor complex and tubulin, can be assembled under cell-free conditions by incubating GR immune pellets with paclitaxel/GTP-stabilized cytosol prepared from GR ؊ L cells. This is the first evidence that the movement of a steroid receptor is dynein-dependent, and it is the first isolation of a steroid receptor bound to the entire system that determines its retrograde movement.As the initial step in their action, transcription factors, such as steroid receptors, p53, and HSF1, must move in a targeted manner through the cytoplasm to the nucleus. Until recently, there has been little mechanistic understanding of how protein solutes (i.e. non-vesicle-associated proteins) undergo such retrograde trafficking. Because the glucocorticoid receptor (GR)

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Isolation and characterization of a mouse L cell variant deficient in glucocorticoid receptors

Cited by 34 publications

References 29 publications

Mechanism of the Anti-Inflammatory Effect of Thiazolidinediones: Relationship with the Glucocorticoid Pathway

Mechanism of the Anti-Inflammatory Effect of Thiazolidinediones: Relationship with the Glucocorticoid Pathway

VBP15, a novel anti‐inflammatory and membrane‐stabilizer, improves muscular dystrophy without side effects

Evidence for Glucocorticoid Receptor Transport on Microtubules by Dynein

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