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Roussel UCLAF (B.M.V., S.D., A.C., F.P., T.G., C.M., M.R.-R.)
93235 Romainville Cedex, France
Institut de
Génétique et de Biologie Moléculaire et Cellulaire
(H.G.) IGBMC-BP. 163 67404 Illkirch Cedex, C.U. de Strasbourg,
France
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ABSTRACT |
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 TOP ABSTRACT INTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
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INTRODUCTION |
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
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Apart from acting as transactivators, several nuclear receptors, including GR, are able to negatively regulate transcription. For GR, two distinct mechanisms have been described. One implies a competition between GR and other transcription factors for binding to their cognate DNA elements (6, 7). The second, generally referred to as transrepression, is much less well understood. It refers to the original observation that transactivation by AP-1 was impaired in the presence of glucocorticoids. Reciprocally, glucocorticoid action was inhibited by AP-1 (Refs. 8, 9, 10, 11, 12 ; reviewed in Ref. 13). Although evidence for direct interaction between GR and AP-1 was provided from in vitro studies indicating that GR and AP-1 mutually interfere with each other’s DNA- binding ability (8, 9, 12), genomic footprinting experiments demonstrated that in the presence of glucocorticoids, AP-1 remains bound to the collagenase promoter in vivo (14). Together with the observation of cell- and promoter-specific transrepression, these results suggested that AP-1 and GR interact rather indirectly, possibly through other transcriptional factors (Ref. 15 ; reviewed in Ref. 13). Mutational studies have demonstrated that both GR DNA- and ligand-binding domains (LBDs) are required for AP-1 transrepression (9, 10, 11, 12, 15, 16). Furthermore, by mutating individual amino acids of the DNA-binding domain of GR, it could be demonstrated that transactivation and transrepression are two separable functions (17). A similar conclusion was drawn for retinoic acid receptors since certain synthetic retinoids elicited AP-1 inhibition without significantly transactivating cognate reporter genes (18, 19, 20).
Glucocorticoids can also repress members of the NF-
B-Rel
transcription factor family (21, 22, 23). In addition to a cross-coupling
mechanism of inhibition between NF-B and the GR, induction of
transcription of the gene encoding IB
has also been recently
demonstrated in lymphocytes and monocytes (24, 25). However, induction
of IB expression by glucocorticoids seems to be cell
type-restricted since it cannot account for inhibition of NF-B
activity in endothelial cells (26).
Glucocorticoids are highly potent antiinflammatory and immunosuppressive agents, due to their pleiotropic effects on the expression/activity of multiple immunomodulators and their ability to induce apoptosis in lymphocytes (Ref. 27 and references therein). However, their therapeutic use is limited by severe side effects, especially during long-term treatment (see Discussion). Whether the dissociation of glucocorticoid-dependent transactivation and transrepression may provide the possibility to separate (some of the) negative side effects from the beneficial antiinflammatory action of classic glucocorticoids is still not clear. Therefore, we have systematically compared the transactivation and transrepression abilities of various glucocorticoid agonists and antagonists and screened a series of synthetic compounds to detect those exhibiting dissociated characteristics (i.e. mainly transactivating or AP-1 transrepressing). Here we report the identification of a novel class of synthetic glucocorticoids that inhibit transcription of the collagenase promoter in transfected Hela cells, while only weakly activating GRE-based reporter genes. Moreover, we show that these ligands are potent inhibitors of interleukin-1ß (IL-1ß) secretion in activated monocytes, while they are unable to induce significant tyrosine amino transferase (TAT) activity in rat HTC hepatoma cells. Finally, we provide evidence that these dissociated glucocorticoids act as antiinflammatory and immunosuppressive drugs in vivo.
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RESULTS |
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
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); we refer to this
class of glucocorticoids as "symmetrical" compounds. All tested
glucocorticoids currently used in medical therapies as antiinflammatory
agents were found to belong to this class (data available upon
request). Each compound was classified relative to the maximal
transactivation and AP-1 transrepression obtained with Dex (100%) and
characterized by its relative activation and repression activities at a
given concentration (Table I, and data not shown).
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. For example, 1 µM RU24858
activated transcription from the GRE5-tk-CAT reporter only
25% as efficiently as the same concentration of Dex (Fig. 1A
), but was similarly potent (85%) in repressing
AP-1-induced transcription from the collagenase promoter (Fig. 1B).
Note that the IC50 of RU24858 for AP-1 transrepression was
20 nM, close to that of prednisolone (Fig. 1B). Two other
compounds, RU24782 and RU40066, exhibited similar dissociated
transactivation and AP-1 transrepression characteristics (see Table 1 and Fig. 1; note that the results given in Table 1
correspond to 100 nM of the indicated ligand). In
vitro competition assays with GR-bound [3H]RU28362,
a pure glucocorticoid agonist (28), or [3H]Dex revealed
that RU24858 and RU24782 bound to GR with a similar relative affinity
as Dex, while RU40066 displayed a 3.7-fold lower affinity (Table 1 and
data not shown). In view of their low agonistic potential and high
affinity to GR, we expected the three dissociated glucocorticoids to
act as efficient antagonists of Dex-induced transactivation. Indeed,
RU40066, at 100 and 1000 nM, efficiently antagonized both
10 nM Dex-induced GRE-tk-CAT (transient transfection) and
TAT (non-transfected HTC cells) activities (Fig. 3).
Surprisingly, however, a 100-fold excess of RU24858 was unable to
antagonize the same Dex-induced activities (Fig. 3), although in
vitro it exhibited an even higher affinity to GR than Dex (Table 1). In these experiments RU24782 was able to partially antagonize the
Dex-induced activities. As in vitro binding assays, RU24858,
RU24782, and RU40066 were shown to compete with
[3H]RU28362; we also measured transcriptional activation
of the GRE-tk-CAT reporter and induction of the TAT activity in Hela
and HTC cells, respectively, exposed to 5 or 10 nM RU28362
with increasing amounts of the dissociated compounds. Again, RU40066
was able to antagonize RU28362-induced transactivation and RU24782
displayed partial antagonist activity, whereas a 100- or 500-fold
excess of RU24858 was unable to antagonize RU28362-induced activity
(data not shown). Thus, apparently dissociated glucocorticoids with a
prevalent potency for AP-1 repression can be designed which are nearly
devoid of both agonistic and antagonistic activities.
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; and data not
shown). Prednisolone (1 µM) induced TAT to 93% of the
stimulation observed with 1 µM Dex (Fig. 4A).
Importantly, only a very weak induction of TAT activity was observed in
HTC cells treated with the different dissociated glucocorticoids, in
keeping with the transactivation results obtained in Hela cells
transiently transfected with GRE5-tk-CAT (Figs. 1A and 4A).
For example, at 1 µM, RU24858 and RU24782 induced TAT
activity to 11% and 17%, respectively, of the stimulation seen with 1
µM Dex. RU40066, like the antagonist RU486, did not
significantly induce TAT activity (Fig. 4A).
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(TNF). Dex, which at 1 µM inhibited 75% of
LPS-induced IL-1ß secretion (not shown), was taken as the reference
compound (-100% in Fig. 4B). Note that the same IC50
values (2 nM) were observed for the Dex-dependent
inhibition of LPS-induced IL-1ß secretion from THP1 cells and the
c-Jun-induced collagenase promoter activity in transiently transfected
Hela cells (Fig. 1, and data not shown). At 1 µM, the
dissociated glucocorticoids RU24782 and RU24858 inhibited IL-1ß
expression 70% and 95%, respectively, of the maximum inhibition
obtained with Dex (Fig. 4B). RU40066 was inactive at doses up to 10
nM and displayed moderate activity (65%) at high
concentrations only, with an IC50 of 0.3 µM
(Fig. 4B, and data not shown). As was the case for Dex, the
IC50 values for inhibition of IL-1ß secretion by
prednisolone, RU24858, and RU24782 were very close to those determined
for AP-1 transrepression in transiently transfected Hela cells (the
following IC50 values were obtained: prednisolone, 6
nM; RU24858, 30 nM; RU24782, <20
nM; compare with Fig. 1B). It is noteworthy that similar
results were obtained with the dissociated glucocorticoids for the
inhibition of LPS-actived IL-1ß secretion from peripheral blood
mononuclear cells (data not shown).
Dissociated Glucocorticoids Display Antiinflammatory and Thymolytic
Activity in Vivo
Glucocorticoids are major tools in the therapy of inflammatory
disorders. Encouraged by the observation that expression of IL-1ß, an
immunomodulator at the top of the inflammatory cascade, was affected by
dissociated glucocorticoids, we investigated whether these compounds
could also exibit antiinflammatory activity in vivo. Two
classic animal models were selected. In the "cotton pellet
granuloma" test (34) the compounds were given orally to rats using
Dex and prednisolone as reference compounds. RU24858 and RU24782
displayed a similar antiinflammatory activity as shown by the
inhibition of granuloma formation; the corresponding ED50
values were estimated to 7 mg/kg and 5 mg/kg, respectively (Table 2).
Dexamethasone, prednisolone, RU24858, and RU24782 inhibited granuloma
formation up to 80%. Prednisolone exhibited maximal efficiency above 5
mg/kg, whereas the activity of RU24782 in this assay plateaued off
above doses of 10 mg/kg (Fig. 5). Although these
compounds were 50 to 70 times less active than Dex, their activities
were very similar to that of prednisolone (ED50 of 2.5
mg/kg; Table 2); in this test system RU40066 was
inactive at the dose of 10 mg/kg, but was not tested at higher
doses.
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). RU40066 displayed a significant, albeit low activity when applied
topically (Table 2).
Notably, RU486, which displayed some transrepression activity with the
Coll-CAT (but not TRE-tk-CAT; Table 1) reporter, was completely
inactive in the cotton pellet granuloma and croton oil-induced ear
edema models at concentrations up to 100 mg/kg (Table 2). Finally, two
further animal models, the "mouse zymosan paw edema" (36) and
"rat carrageenin paw edema" (37), confirmed that the
antiinflammatory potency of RU24858 was, albeit lower than that of Dex,
similar to that of prednisolone (data not shown).
As the immunosuppressive activity of classic glucocorticoids is linked
to their ability to induce lymphocyte apoptosis, we measured thymolytic
activity of dissociated glucocorticoids in the same rats that were used
for the cotton-pellet granuloma test. The doses at which a reduction of
thymus weights could be observed were generally lower than those needed
for inhibiting granuloma formation (Table 2). RU24782 and RU24858
displayed the same activity (ED50 estimated to 2.5 mg/kg)
which was, as in the granuloma test, near to that of prednisolone (1.6
mg/kg). Doses of 5 mg/kg of RU24858 or RU24782 induced a thymus weight
reduction of 81% and 77%, respectively, compared with control
animals. Notably, even at 1 mg/kg, these glucocorticoids exhibited
significant activity (25% reduction of thymus weight,
P < 0.05), whereas they were inactive at these doses
in the granuloma test (data not shown). As expected, Dex was highly
active in the thymolysis test (with an ED50 below 0.05
mg/kg) since at this lowest dose tested, the decrease of thymus weight
was 60% (data not shown). As previously observed in the cotton-pellet
granuloma, RU40066 was inactive at the dose tested (10 mg/kg). Taken
together, in several in vivo animal models, the dissociated
glucocorticoids RU24858 and RU24782, which are weak activators of
positively regulated GR target genes, displayed consistently a strong
antiinflammatory action that was similar, or even superior, to
that of the classic antiinflammatory glucocorticoid prednisolone.
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DISCUSSION |
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
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With the aim of better understanding how glucocorticoids enable GR to
transrepress AP-1 activity (see Introduction for references), we have
compared AP-1 inhibition in the presence of the three above types of
ligands and, moreover, screened a library of glucocortcoids to find
ligands that would inhibit AP-1 activity without significantly inducing
agonistic activity (termed "dissociated" glucocorticoids).
Confirming the original data of Jonat et al. (9), we
observed that in transfected HeLa cells the type I antiglucocorticoid
RU38486 was unable to repress AP-1-induced transactivation of a
TRE5-tk-CAT reporter (Table 1), and, indeed, could reverse
the effect of the agonist Dex (data not shown). With a collagenase
promoter-based reporter, some RU38486-dependent AP-1 repression was
observed but the antagonist still reversed the repression induced by 10
nM Dex (Fig. 6). No transrepressing ability
was conferred onto GR in the presence of the type II antagonist RU43044
with any of the two AP-1 reporters (Table 1). Note that in the above
cases transrepression was via the endogenous GR; transiently expressed
exogenous GR can apparently further increase the AP-1 inhibition (17)
but leads to nonphysiological GR concentrations. In conclusion, neither
type I nor type II antiglucocorticoids have the characteristics of
dissociated glucocorticoids and, thus, have only limited promise for
use as antiinflammatory agents (see below).
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). Note in this context that
prednisolone, widely used as antiinflammatory glucocorticoid, showed a
similar anti-AP-1 activity in transfected cells (Table 1). The most
interesting of these compounds is RU24858, since it exemplifies a novel
class of glucocorticoids: despite a binding affinity that is even
higher than that of Dex, RU24858 does not significantly antagonize
Dex-induced transcription (Fig. 3). However, in in vitro
binding assays RU24858 does efficiently compete with radiolabeled Dex
(and other glucocorticoid agonists) for binding to the GR. This implies
that, in contrast to classic glucocorticoids, the binding affinity of
RU24858 to transcriptionally engaged GR in vivo is lower
than that measured in vitro. Importantly, however, this
lower in vivo binding affinity does not significantly
decrease the ability of the GR-RU24858 complex to repress AP-1
activity. We conclude from these data that AP-1 repression involves
very specific ligand-induced structural alterations of GR. How can RU24858 bind GR with different affinities in vitro and in vivo? Transcriptional interference/squelching studies have suggested that transcriptional intermediary factors (TIFs, also termed mediators, coactivators, bridging factors) mediate the ligand-dependent activation function AF-2 of steroid receptors to the transcriptional machinery (45, 46, 47). Putative TIFs have been recently isolated and characterized on the basis of their ligand-dependent interaction with several nuclear receptors (48, 49, 50, 51, 52, 53, 54). We propose that the interaction between the LBD and TIF(s) (or any other factor that interacts with the GR LBD in vivo) may differentially affect the binding of Dex and RU24858, resulting in a decreased affinity of RU24858, but not Dex, to the GR-TIF complex. However, noncomplexed GRs, or a set of GR complexes distinct from those involved in transactivation, may bind RU24858 with high affinity and give rise to the strong AP-1 inhibition observed in the presence of this compound. Note that there is a precedent for an altered affinity of glucocorticoids upon GR complex formation, since different affinities have been reported for the GR and the GR-hsp90 complex (55, 56).
Taken together, the above results suggest that we have identified a novel class of glucocorticoids that are neither efficient agonists nor antagonists in vivo, but can very efficiently induce GR-dependent AP-1 repression.
In Contrast to Classic Antiglucocorticoids, Dissociated
Glucocorticoids Are Strong Inhibitors of LPS-Induced IL-1ß
Secretion
Glucocorticoids are powerful antiinflammatory agents, most likely
due to their ability to block the expression of multiple cytokines.
Inhibition of cytokine expression has been reported to occur at the
transcriptional level for IL-1 to IL-6, IL-8, TNF,
colony-stimulating factor (CSF)-1/macrophage (M)-CSF, granulocyte
macrophage (GM)-CSF, and 
-interferon (IL-1 expression is blocked at
other levels as well; TNF and GM-CSF expression may be blocked through
degradation of their mRNAs; for reviews see Refs. 27, 57, 58, 59, 60 , and
references therein). Most of these genes require transcription factors,
such as AP-1, NF-B, or NF-AT, for their expression, suggesting that
glucocorticoids exert their antiinflammatory functions by negatively
interfering with the activity of (some of) these factors. Dissociated
glucocorticoids, such as RU24858, were as potent inhibitors of IL-1ß
secretion as prednisolone in LPS-stimulated cultured human monocytes,
whereas classic antiglucocorticoids with no (RU43044) or
promoter-dependent AP-1-transrepression activities (RU486)
inhibited IL-1ß secretion only weakly (RU486) or not at all (RU43044)
in this assay (Fig. 4 and data not shown).
In Vivo Dissociated Glucocorticoids Are
Antiinflammatory Agents as Potent as Prednisolone
To investigate whether dissociated glucocorticoids could be
active as antiinflammatory drugs in vivo, we studied two
established animal models, the cotton pellet granuloma model, in which
the glucocorticoid is given orally to rats, and the croton oil-induced
ear edema model, in which the drug is applied topically to mice (see
Materials and Methods for details and references).
Importantly, in these in vivo models, RU24858 was as active
as prednisolone (Fig. 5 and Table 2). In the croton oil-induced ear
edema model, which relies on phorbol ester-induced skin inflammation,
RU24858 was even 2-fold more active than prednisolone, and RU24782 was
about half as active as prednisolone. It is unclear why RU40066 was
inactive; at present, no data on the metabolism and stability in
vivo of this compound are available. Importantly, in keeping with
their inability to inhibit IL-1ß secretion, the classic antagonists
RU486 and RU43044 were completely inactive in these in vivo
tests (Table 2 and data not shown).
The immunosuppressant potential of glucocorticoids relates to
their ability to induce T cell apoptosis and to inhibit cytokine gene
expression. These activities are therapeutically exploited in organ
transplantation and in treatment of leukemias. We compared, in the
cotton pellet granuloma model, the extent of thymolysis induced by
classic (anti)glucocorticoids and the novel class of dissociated
glucocorticoids. Again we observed a high activity of RU24858 and
RU24782, similar to that of prednisolone, while antiglucocorticoids
were inactive (Table 2 and data not shown).
Dissociated Glucocorticoids May be Novel Tools to Improve
Antiinflammatory Glucocorticoid Therapy
We describe here a novel class of dissociated glucocorticoids,
distinct from the various previously reported agonists and antagonists.
These compounds efficiently transrepress AP-1 activity, while only
marginally activating glucocorticoid target genes. The dissociated
glucocorticoids have maintained the antiinflammatory and, according to
our as yet limiting data, also the thymolytic capacity of classical
glucocorticoid agonists. Moreover, its prototypic member RU24858 is as
active, or, depending on the system, even more active, than
prednisolone in standard in vivo models.
Hypothalamic-pituitary-adrenal axis insufficiency, osteoporosis,
diabetes, steroid myopathy, and infectious and neuropsychiatric
complications limit the therapeutic use of classic glucocorticoid
agonists (61). At least some of these complications are likely to
derive from the agonistic activities of the classic glucocorticoids,
and it has not escaped our attention that our glucocorticoid analogs,
provided that the dissociation of transactivation and transrepression
is maintained in target organs, have the potential to constitute a new
class of antiinflammatory glucocorticoids with significantly reduced
side effects. Studies are under way to confirm this hypothesis.
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MATERIALS AND METHODS |
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
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Cell Culture and Transfection
Hela cells were cultured in DMEM supplemented with 10% FCS and
grown at 37 C and 5% CO2. Transient transfection assays
for transactivation or AP-1 transrepression were carried out using the
calcium phosphate coprecipitation procedure. Briefly, 4 x
105 cells were seeded 24 h before transfection in
six-well plates containing DMEM supplemented with 5%
charcoal/dextran-treated serum. For transactivation assays, HeLa cells
were transfected with 1 µg GRE5-tk-CAT reporter, 1 µg
polyIIßGal, and 3 µg Bluescript KS (Stratagene, La Jolla, CA). For
transrepression assays, HeLa cells were transfected with 3 µg of the
Coll(-517/+63)CAT reporter, 1 µg polyIIßGal, 250 ng pSVc-Jun
expression plasmid, and 1 µg Bluescript KS. Sixteen hours later,
cells were rinsed with Earle’s Balanced Salt Solution (EBSS), the
medium was replaced, and the hormones were added as indicated. After
24 h, cells were washed with ice-cold EBSS and resuspended in 250
µl of lysis buffer provided with the chloramphenicol
acetyltransferase enzyme-linked immunosorbent assay (CAT ELISA) kit
(Boehringer Mannheim, Indianapolis, IN). The amount of CAT enzyme was
quantified according to the manufacturer’s instructions. Results were
normalized according to the ß-galactosidase activity originating from
cotransfected polyIIßGal and plotted relative to the transactivation
seen with 1 µM Dex (=100%) or relative to the AP-1
transrepression exerted by 1 µM Dex (=-100%).
SEMs were calculated from a minimum of three independent
assays per compound and per concentration.
Determination of Relative Binding Affinities
Relative binding affinities were determined by incubating HeLa
cell cytosol for 24 h at 0 C with either [3H]RU28362
(28) or [3H]Dex with or without different concentrations
of competitor steroids. Bound and free ligands were separated by the
dextran-coated charcol method (38). The relative binding affinity of
Dex was taken as reference (100%). Similar results were obtained with
crude SF9 cell extracts containing high levels of human GR expressed
from recombinant baculoviruses.
TAT Assay
HTC cells were cultured in DMEM supplemented with 10% FCS and
kept at 37 C and 5% CO2. For steroid treatment
106 cells per well were seeded into six-well plates. After
24 h, at confluency, the medium was replaced by DMEM without
serum, and hormones were added for 18 h. TAT activity was modified
from Ref. 64 to be measured in 96-well plates. The endogenous enzymatic
activity measured in control cells was subtracted, and the results were
expressed relative to the maximal activity measured in cells treated
with 10-6 M Dex.
Assay of IL-1ß Secretion
The monocytic cell line THP1 (ATCC: TIB202) was cultured in RPMI
1640 supplemented with 10% FCS and grown at 37 C and 5%
CO2. IL-1ß secretion was induced by treating the cells
with 5 µg/ml LPS (Sigma, St. Louis, MO); glucocorticoids were added
simultaneously. After 18 h, supernatants were collected and
IL-1ß was quantitated using the human IL-1ß ELISA kit from R&D
Systems (Minneapolis, MN).
Antiinflammatory Activity in the Cotton-Pellet Granuloma Test
The cotton-pellet granuloma assay was performed as described
previously (34). Two cotton pellets (10 mg each) were inserted
subcutaneously into the upper dorsal area of female Wistar rats (weight
range 90–100 g, Iffa Credo, France). Test compounds were administered
orally, once a day, for 4 days. Twenty-four hours after the last
treatment, the animal was killed and the pellet, along with the
surrounding granuloma, was carefully dissected from the animal and its
dry weight was determined. For this, granuloma and pellet were heated
at 60 C overnight. By subtracting the initial weight of the pellet, the
dry weight of the granuloma was determined. The activities of the
various synthetic glucocorticoids were expressed as ED50
values (dose causing a reduction of granuloma weight by 50%).
Thymolytic Activity
The thymolytic activity of the synthetic compounds was
determined by weighing the thymus of the rats treated in the granuloma
test. The ED50 values correspond to the glucocorticoid
doses inducing a 50% decrease in thymus weights.
Topical Antiinflammatory Effect on the Croton Oil-Induced Ear
Edema
This test was carried out as described previously (35) on groups
of eight male OF1 mice weighing 18–22 g (Iffa Credo, L’Arbresle,
France). The edema was induced on one ear by the application of a
solution of croton oil (2% vol/vol) in pyridine-water-ether 4:1:14.6
(by volume). Animals were killed 6 h later, and the ears were
removed and weighed. Edema was determined from the difference in weight
between the irritant-treated and the contralateral ear. The compounds
to be tested were dissolved in the croton oil solution and topically
applied on the ear. ED50 values correspond to the doses
reducing the control edema by 50%.
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ACKNOWLEDGMENTS |
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FOOTNOTES |
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1 Present address: INSERM U 248, Institut Curie, 26 rue d’Ulm, 75231
Paris Cedex 05. 
2 S. Dupont is currently working at Institut de Génétique
et de Biologie Moléculaire et Cellulaire.
Received for publication November 27, 1996. Revision received May 14, 1997. Accepted for publication May 19, 1997.
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G. Hochhaus New Developments in Corticosteroids Proceedings of the ATS, November 1, 2004; 1(3): 269 - 274. [Abstract] [Full Text] [PDF]
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 C. Desmet, P. Gosset, B. Pajak, D. Cataldo, M. Bentires-Alj, P. Lekeux, and F. Bureau Selective Blockade of NF-{kappa}B Activity in Airway Immune Cells Inhibits the Effector Phase of Experimental Asthma J. Immunol., November 1, 2004; 173(9): 5766 - 5775. [Abstract] [Full Text] [PDF]
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 O. Kassel, S. Schneider, C. Heilbock, M. Litfin, M. Gottlicher, and P. Herrlich A nuclear isoform of the focal adhesion LIM-domain protein Trip6 integrates activating and repressing signals at AP-1- and NF-{kappa}B-regulated promoters Genes & Dev., October 15, 2004; 18(20): 2518 - 2528. [Abstract] [Full Text] [PDF]
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 L. E. Donnelly, R. Newton, G. E. Kennedy, P. S. Fenwick, R. H. F. Leung, K. Ito, R. E. K. Russell, and P. J. Barnes Anti-inflammatory effects of resveratrol in lung epithelial cells: molecular mechanisms Am J Physiol Lung Cell Mol Physiol, October 1, 2004; 287(4): L774 - L783. [Abstract] [Full Text] [PDF]
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H. Schacke, A. Schottelius, W.-D. Docke, P. Strehlke, S. Jaroch, N. Schmees, H. Rehwinkel, H. Hennekes, and K. Asadullah Dissociation of transactivation from transrepression by a selective glucocorticoid receptor agonist leads to separation of therapeutic effects from side effects PNAS, January 6, 2004; 101(1): 227 - 232. [Abstract] [Full Text] [PDF]
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 I. Kurucz, S. Toth, K. Nemeth, K. Torok, V. Csillik-Perczel, A. Pataki, C. Salamon, Z. Nagy, J. I. Szekely, K. Horvath, et al. Potency and Specificity of the Pharmacological Action of a New, Antiasthmatic, Topically Administered Soft Steroid, Etiprednol Dicloacetate (BNP-166) J. Pharmacol. Exp. Ther., October 1, 2003; 307(1): 83 - 92. [Abstract] [Full Text] [PDF]
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 P. J. Barnes and I. M. Adcock How Do Corticosteroids Work in Asthma? Ann Intern Med, September 2, 2003; 139(5_Part_1): 359 - 370. [Full Text] [PDF]
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K. De Bosscher, W. Vanden Berghe, and G. Haegeman The Interplay between the Glucocorticoid Receptor and Nuclear Factor-{kappa}B or Activator Protein-1: Molecular Mechanisms for Gene Repression Endocr. Rev., August 1, 2003; 24(4): 488 - 522. [Abstract] [Full Text] [PDF]
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A. Stevens, H. Garside, A. Berry, C. Waters, A. White, and D. Ray Dissociation of Steroid Receptor Coactivator 1 and Nuclear Receptor Corepressor Recruitment to the Human Glucocorticoid Receptor by Modification of the Ligand-Receptor Interface: The Role of Tyrosine 735 Mol. Endocrinol., May 1, 2003; 17(5): 845 - 859. [Abstract] [Full Text] [PDF]
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M. J. Coghlan, P. B. Jacobson, B. Lane, M. Nakane, C. W. Lin, S. W. Elmore, P. R. Kym, J. R. Luly, G. W. Carter, R. Turner, et al. A Novel Antiinflammatory Maintains Glucocorticoid Efficacy with Reduced Side Effects Mol. Endocrinol., May 1, 2003; 17(5): 860 - 869. [Abstract] [Full Text] [PDF]
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Y. He, D. Szapary, and S. S. Simons Jr. Modulation of Induction Properties of Glucocorticoid Receptor-Agonist and -Antagonist Complexes by Coactivators Involves Binding to Receptors but Is Independent of Ability of Coactivators to Augment Transactivation J. Biol. Chem., December 13, 2002; 277(51): 49256 - 49266. [Abstract] [Full Text] [PDF]
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 M. Lasa, S. M. Abraham, C. Boucheron, J. Saklatvala, and A. R. Clark Dexamethasone Causes Sustained Expression of Mitogen-Activated Protein Kinase (MAPK) Phosphatase 1 and Phosphatase-Mediated Inhibition of MAPK p38 Mol. Cell. Biol., November 15, 2002; 22(22): 7802 - 7811. [Abstract] [Full Text] [PDF]
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R. Korhonen, A. Lahti, M. Hamalainen, H. Kankaanranta, and E. Moilanen Dexamethasone Inhibits Inducible Nitric-Oxide Synthase Expression and Nitric Oxide Production by Destabilizing mRNA in Lipopolysaccharide-Treated Macrophages Mol. Pharmacol., September 1, 2002; 62(3): 698 - 704. [Abstract] [Full Text] [PDF]
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C. W. Lin, M. Nakane, M. Stashko, D. Falls, J. Kuk, L. Miller, R. Huang, C. Tyree, J. N. Miner, J. Rosen, et al. trans-Activation and Repression Properties of the Novel Nonsteroid Glucocorticoid Receptor Ligand 2,5-Dihydro-9-hydroxy-10-methoxy-2,2,4-trimethyl-5-(1-methylcyclohexen-3-y1)-1H-[1]benzopyrano[3,4-f]quinoline (A276575) and Its Four Stereoisomers Mol. Pharmacol., August 1, 2002; 62(2): 297 - 303. [Abstract] [Full Text] [PDF]
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J. Chen, S. Kaul, and S. S. Simons Jr. Structure/Activity Elements of the Multifunctional Protein, GMEB-1. CHARACTERIZATION OF DOMAINS RELEVANT FOR THE MODULATION OF GLUCOCORTICOID RECEPTOR TRANSACTIVATION PROPERTIES J. Biol. Chem., June 7, 2002; 277(24): 22053 - 22062. [Abstract] [Full Text] [PDF]
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N. Yoshikawa, Y. Makino, K. Okamoto, C. Morimoto, I. Makino, and H. Tanaka Distinct Interaction of Cortivazol with the Ligand Binding Domain Confers Glucocorticoid Receptor Specificity. CORTIVAZOL IS A SPECIFIC LIGAND FOR THE GLUCOCORTICOID RECEPTOR J. Biol. Chem., February 8, 2002; 277(7): 5529 - 5540. [Abstract] [Full Text] [PDF]
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J. D. Baxter, P. Goede, J. W. Apriletti, B. L. West, W. Feng, K. Mellstrom, R. J. Fletterick, R. L. Wagner, P. J. Kushner, R. C. J. Ribeiro, et al. Structure-Based Design and Synthesis of a Thyroid Hormone Receptor (TR) Antagonist Endocrinology, February 1, 2002; 143(2): 517 - 524. [Abstract] [Full Text] [PDF]
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T. Miura, R. Ouchida, N. Yoshikawa, K. Okamoto, Y. Makino, T. Nakamura, C. Morimoto, I. Makino, and H. Tanaka Functional Modulation of the Glucocorticoid Receptor and Suppression of NF-kappa B-dependent Transcription by Ursodeoxycholic Acid J. Biol. Chem., December 7, 2001; 276(50): 47371 - 47378. [Abstract] [Full Text] [PDF]
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K.F. Rabe and D.T. Schmidt Pharmacological treatment of asthma today Eur. Respir. J., July 2, 2001; 18(34_suppl): 34S - 40s. [Abstract] [Full Text] [PDF]
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 A. Aranda and A. Pascual Nuclear Hormone Receptors and Gene Expression Physiol Rev, July 1, 2001; 81(3): 1269 - 1304. [Abstract] [Full Text] [PDF]
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 M. KRAFT, Q. HAMID, G. P. CHROUSOS, R. J. MARTIN, and D. Y. M. LEUNG Decreased Steroid Responsiveness at Night in Nocturnal Asthma . Is the Macrophage Responsible? Am. J. Respir. Crit. Care Med., April 1, 2001; 163(5): 1219 - 1225. [Abstract] [Full Text]
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K. De Bosscher, W. Vanden Berghe, and G. Haegeman Glucocorticoid Repression of AP-1 Is Not Mediated by Competition for Nuclear Coactivators Mol. Endocrinol., February 1, 2001; 15(2): 219 - 227. [Abstract] [Full Text]
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M. G. Belvisi, S. L. Wicks, C. H. Battram, S. E. W. Bottoms, J. E. Redford, P. Woodman, T. J. Brown, S. E. Webber, and M. L. Foster Therapeutic Benefit of a Dissociated Glucocorticoid and the Relevance of In Vitro Separation of Transrepression from Transactivation Activity J. Immunol., February 1, 2001; 166(3): 1975 - 1982. [Abstract] [Full Text] [PDF]
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 M. V. Gonzalez, B. Jimenez, M. T. Berciano, J. M. Gonzalez-Sancho, C. Caelles, M. Lafarga, and A. Munoz Glucocorticoids Antagonize Ap-1 by Inhibiting the Activation/Phosphorylation of Jnk without Affecting Its Subcellular Distribution J. Cell Biol., September 4, 2000; 150(5): 1199 - 1208. [Abstract] [Full Text] [PDF]
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D. JAFFUEL, P. DEMOLY, C. GOUGAT, P. BALAGUER, G. MAUTINO, P. GODARD, J. BOUSQUET, and M. MATHIEU Transcriptional Potencies of Inhaled Glucocorticoids Am. J. Respir. Crit. Care Med., July 1, 2000; 162(1): 57 - 63. [Abstract] [Full Text]
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 R. Newton Molecular mechanisms of glucocorticoid action: what is important? Thorax, July 1, 2000; 55(7): 603 - 613. [Full Text]
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K. De Bosscher, W. Vanden Berghe, L. Vermeulen, S. Plaisance, E. Boone, and G. Haegeman Glucocorticoids repress NF-kappa B-driven genes by disturbing the interaction of p65 with the basal transcription machinery, irrespective of coactivator levels in the cell PNAS, April 11, 2000; 97(8): 3919 - 3924. [Abstract] [Full Text] [PDF]
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R. M. Sapolsky, L. M. Romero, and A. U. Munck How Do Glucocorticoids Influence Stress Responses? Integrating Permissive, Suppressive, Stimulatory, and Preparative Actions Endocr. Rev., February 1, 2000; 21(1): 55 - 89. [Abstract] [Full Text]
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D. Luo, E. Guerin, M.-G. Ludwig, I. Stoll, P. Basset, and P. Anglard Transcriptional Induction of Stromelysin-3 in Mesodermal Cells Is Mediated by an Upstream CCAAT/Enhancer-binding Protein Element Associated with a DNase I-hypersensitive Site J. Biol. Chem., December 24, 1999; 274(52): 37177 - 37185. [Abstract] [Full Text] [PDF]
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 K Thiele, A Bierhaus, F Autschbach, M Hofmann, W Stremmel, H Thiele, R Ziegler, and P P Nawroth Cell specific effects of glucocorticoid treatment on the NF-kappa Bp65/Ikappa Balpha system in patients with Crohn's disease Gut, November 1, 1999; 45(5): 693 - 704. [Abstract] [Full Text] [PDF]
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 C. M. Bamberger, T. Else, A.-M. Bamberger, F. Ulrich Beil, and H. M. Schulte Dissociative Glucocorticoid Activity of Medroxyprogesterone Acetate in Normal Human Lymphocytes J. Clin. Endocrinol. Metab., November 1, 1999; 84(11): 4055 - 4061. [Abstract] [Full Text]
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D. W. Ray, C.-S. Suen, A. Brass, J. Soden, and A. White Structure/Function of the Human Glucocorticoid Receptor: Tyrosine 735 Is Important for Transactivation Mol. Endocrinol., November 1, 1999; 13(11): 1855 - 1863. [Abstract] [Full Text]
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W. Vanden Berghe, E. Francesconi, K. De Bosscher, M. Resche-Rigon, and G. Haegeman Dissociated Glucocorticoids with Anti-Inflammatory Potential Repress Interleukin-6 Gene Expression by a Nuclear Factor-kappa B-Dependent Mechanism Mol. Pharmacol., October 1, 1999; 56(4): 797 - 806. [Abstract] [Full Text]
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O. Kassel, F. Schmidlin, C. Duvernelle, F. de Blay, and N. Frossard Up- and Down-Regulation by Glucocorticoids of the Constitutive Expression of the Mast Cell Growth Factor Stem Cell Factor by Human Lung Fibroblasts in Culture Mol. Pharmacol., December 1, 1998; 54(6): 1073 - 1079. [Abstract] [Full Text]
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R. Newton, J. Seybold, L. M. E. Kuitert, M. Bergmann, and P. J. Barnes Repression of Cyclooxygenase-2 and Prostaglandin E2 Release by Dexamethasone Occurs by Transcriptional and Post-transcriptional Mechanisms Involving Loss of Polyadenylated mRNA J. Biol. Chem., November 27, 1998; 273(48): 32312 - 32321. [Abstract] [Full Text] [PDF]
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 S. Wakisaka, N. Suzuki, M. Takeno, Y. Takeba, H. Nagafuchi, N. Saito, H. Hashimoto, T. Tomita, T. Ochi, and T. Sakane Involvement of simultaneous multiple transcription factor expression, including cAMP responsive element binding protein and OCT-1, for synovial cell outgrowth in patients with rheumatoid arthritis Ann Rheum Dis, August 1, 1998; 57(8): 487 - 494. [Abstract] [Full Text]
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K. Ito, E. Jazrawi, B. Cosio, P. J. Barnes, and I. M. Adcock p65-activated Histone Acetyltransferase Activity Is Repressed by Glucocorticoids. MIFEPRISTONE FAILS TO RECRUIT HDAC2 TO THE p65-HAT COMPLEX J. Biol. Chem., August 3, 2001; 276(32): 30208 - 30215. [Abstract] [Full Text] [PDF]
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