Faculty Opinions recommendation of NALP3 inflammasome upregulation and CASP1 cleavage of the glucocorticoid receptor cause glucocorticoid resistance in leukemia cells.

We determined the mol wt of glucocorticoid receptors in human leukemia cells in order to detect glucocorticoid receptor defects that might cause glucocorticoid resistance. Glucocorticoid receptors in intact cells were affinity labeled with [3H]dexamethasone-21-mesylate and were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Receptors in normal human peripheral blood mononuclear cells and six human leukemia cell lines had mol wt of 97,000. Malignant cells from ten of 25 patients with leukemia contained electrophoretically abnormal glucocorticoid receptors having mol wt of 55,000 in addition to normal size receptors (Mr = 97,000). The receptor abnormality was not restricted to a particular type of leukemia and was seen in cells from both newly diagnosed patients and patients who had received prior chemotherapy, including prednisone. The abnormal receptor was not generated when cells having only normal size receptors were assayed under conditions that favor proteolysis or when cytosol from cells containing the abnormal receptor form was mixed with cytosol from cells containing only normal size receptors. The mol wt of the abnormal receptors in human leukemia cells was the same as the mol wt of receptors in mutant mouse lymphoma cell lines, S49 143R and S49 55R, which have the nuclear transfer-increased phenotype of glucocorticoid resistance. This work describes for the first time a single common electrophoretic abnormality of glucocorticoid receptors in human leukemia cells. Further investigation of glucocorticoid receptor defects in human leukemia cells could lead to an improved understanding of the mechanisms of glucocorticoid resistance in leukemia as well as a method of predicting which patients are likely to be resistant to glucocorticoid therapy.

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A single common electrophoretic abnormality of glucocorticoid receptors in human leukemia cells

Blood ◽

10.1182/blood.v66.3.679.679 ◽

1985 ◽

Vol 66 (3) ◽

pp. 679-685 ◽

Cited By ~ 9

Author(s):

CW Distelhorst ◽

BM Benutto ◽

RC Griffith

Keyword(s):

Glucocorticoid Receptor ◽

Cell Lines ◽

Glucocorticoid Receptors ◽

Sodium Dodecyl ◽

Leukemia Cell ◽

Glucocorticoid Resistance ◽

Leukemia Cells ◽

Human Leukemia ◽

Intact Cells ◽

Human Leukemia Cells

Abstract We determined the mol wt of glucocorticoid receptors in human leukemia cells in order to detect glucocorticoid receptor defects that might cause glucocorticoid resistance. Glucocorticoid receptors in intact cells were affinity labeled with [3H]dexamethasone-21-mesylate and were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Receptors in normal human peripheral blood mononuclear cells and six human leukemia cell lines had mol wt of 97,000. Malignant cells from ten of 25 patients with leukemia contained electrophoretically abnormal glucocorticoid receptors having mol wt of 55,000 in addition to normal size receptors (Mr = 97,000). The receptor abnormality was not restricted to a particular type of leukemia and was seen in cells from both newly diagnosed patients and patients who had received prior chemotherapy, including prednisone. The abnormal receptor was not generated when cells having only normal size receptors were assayed under conditions that favor proteolysis or when cytosol from cells containing the abnormal receptor form was mixed with cytosol from cells containing only normal size receptors. The mol wt of the abnormal receptors in human leukemia cells was the same as the mol wt of receptors in mutant mouse lymphoma cell lines, S49 143R and S49 55R, which have the nuclear transfer-increased phenotype of glucocorticoid resistance. This work describes for the first time a single common electrophoretic abnormality of glucocorticoid receptors in human leukemia cells. Further investigation of glucocorticoid receptor defects in human leukemia cells could lead to an improved understanding of the mechanisms of glucocorticoid resistance in leukemia as well as a method of predicting which patients are likely to be resistant to glucocorticoid therapy.

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Glucocorticoid receptor heterozygosity combined with lack of receptor auto-induction causes glucocorticoid resistance in Jurkat acute lymphoblastic leukemia cells

Cell Death and Differentiation ◽

10.1038/sj.cdd.4401413 ◽

2004 ◽

Vol 11 (S1) ◽

pp. S65-S72 ◽

Cited By ~ 37

Author(s):

S Riml ◽

S Schmidt ◽

M J Ausserlechner ◽

S Geley ◽

R Kofler

Keyword(s):

Acute Lymphoblastic Leukemia ◽

Glucocorticoid Receptor ◽

Lymphoblastic Leukemia ◽

Glucocorticoid Resistance ◽

Leukemia Cells

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Glucocorticoid resistance conferring mutation in the C-terminus of GR alters the receptor conformational dynamics

Scientific Reports ◽

10.1038/s41598-021-92039-9 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Anna Kaziales ◽

Florian Rührnößl ◽

Klaus Richter

Keyword(s):

Glucocorticoid Receptor ◽

Steroid Hormones ◽

In Silico ◽

Conformational Dynamics ◽

Glucocorticoid Resistance ◽

Physiological Processes ◽

C Terminus ◽

Hsp90 Chaperone ◽

In Silico Methods

AbstractThe glucocorticoid receptor is a key regulator of essential physiological processes, which under the control of the Hsp90 chaperone machinery, binds to steroid hormones and steroid-like molecules and in a rather complicated and elusive response, regulates a set of glucocorticoid responsive genes. We here examine a human glucocorticoid receptor variant, harboring a point mutation in the last C-terminal residues, L773P, that was associated to Primary Generalized Glucocorticoid Resistance, a condition originating from decreased affinity to hormone, impairing one or multiple aspects of GR action. Using in vitro and in silico methods, we assign the conformational consequences of this mutation to particular GR elements and report on the altered receptor properties regarding its binding to dexamethasone, a NCOA-2 coactivator-derived peptide, DNA, and importantly, its interaction with the chaperone machinery of Hsp90.

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Neutrophil elastase produces 52-kD and 30-kD glucocorticoid receptor fragments in the cytosol of human leukemia cells

Blood ◽

10.1182/blood.v70.3.860.bloodjournal703860 ◽

1987 ◽

Vol 70 (3) ◽

pp. 860-868

Author(s):

CW Distelhorst ◽

KE Janiga ◽

KJ Howard ◽

SE Strandjord ◽

EJ Campbell

Keyword(s):

Glucocorticoid Receptor ◽

Neutrophil Elastase ◽

Glucocorticoid Receptors ◽

Leukemia Cell ◽

Specific Inhibitor ◽

Leukemia Cells ◽

Human Leukemia ◽

Receptor Fragments ◽

Limited Digestion

Characterization of glucocorticoid receptors in leukemia cells is important to understand mechanisms of glucocorticoid resistance but has been impeded by receptor fragmentation in cytosol extracts. We recently found that formation of 52- and 30-kilodalton (kD) glucocorticoid receptor fragments in cytosol of leukemia cells is due to proteolysis and is blocked by diisopropylfluorophosphate (DFP). In the present study, we identify a 28-kD serine protease in cytosol of leukemia cells that binds [3H]DFP and correlates with the formation of 52- and 30-kD receptor fragments. This protease is immunoprecipitated by antiserum to neutrophil elastase. Limited digestion of [3H]dexamethasone-21-mesylate- labeled receptors by purified neutrophil elastase produces 52- and 30- kD receptor fragments. Receptor fragmentation in the cytosol of leukemia cells in inhibited by methoxysuccinyl-alanyl-alanyl-prolyl- valyl-chloromethylketone, a highly specific inhibitor of neutrophil elastase. The addition of as few as 5% neutrophils to a lymphoid cell suspension provides sufficient elastase to produce receptor fragmentation. Our findings indicate that neutrophil elastase is responsible for receptor fragmentation in the cytosol of leukemia cells. The neutrophil elastase may be endogenous to the leukemia cells or may come from neutrophils that contaminate leukemia cell suspensions.

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