Abnormal glucocorticoid receptors in acute leukemia cells

In normal tissues, 3H-triamcinolone acetonide (3H-TA) labeled glucocorticoid receptors can be resolved into 2 components by DEAE chromatography: peak I elutes at 0.04 M salt and peak II at 0.22 M salt. By glycerol gradient centrifugation, peak I is 3.5S and peak II is 8.5S. Peak I binds to DNA, while peak II does not. Blast cell 3H-TA- binding macromolecules in 27 of 62 cases of acute leukemia had DEAE binding characteristics identical to those of normal tissues; the remaining 35 cases were abnormal. In these cases there was either a single DEAE species eluting in the peak I area (30 cases) or multiple low-amplitude peaks eluting across the entire gradient (5 cases). The abnormal single peak material failed to bind to DNA in 5 cases (of 5 studied), whereas peak I material from 5 cases (of 5 studied), showing normal peak I-peak II ratios, bound normally to DNA. In 3 cases (of 3 studied), the abnormal single peak material had an S value of 2–2.5S, whereas in 5 cases with normal peak I-peak II ratios, the S values were 3.5S and 8.5S, respectively. We hypothesize that those leukemias with abnormal binder characteristics cannot respond to glucocorticoid therapy.

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Abnormal glucocorticoid receptors in acute leukemia cells

Blood ◽

10.1182/blood.v59.2.393.393 ◽

1982 ◽

Vol 59 (2) ◽

pp. 393-400 ◽

Cited By ~ 16

Author(s):

R McCaffrey ◽

A Lillquist ◽

R Bell

Keyword(s):

Acute Leukemia ◽

Triamcinolone Acetonide ◽

Glucocorticoid Receptors ◽

Gradient Centrifugation ◽

Blast Cell ◽

Leukemia Cells ◽

Single Peak ◽

Binding Characteristics ◽

Normal Tissues ◽

Low Amplitude

Abstract In normal tissues, 3H-triamcinolone acetonide (3H-TA) labeled glucocorticoid receptors can be resolved into 2 components by DEAE chromatography: peak I elutes at 0.04 M salt and peak II at 0.22 M salt. By glycerol gradient centrifugation, peak I is 3.5S and peak II is 8.5S. Peak I binds to DNA, while peak II does not. Blast cell 3H-TA- binding macromolecules in 27 of 62 cases of acute leukemia had DEAE binding characteristics identical to those of normal tissues; the remaining 35 cases were abnormal. In these cases there was either a single DEAE species eluting in the peak I area (30 cases) or multiple low-amplitude peaks eluting across the entire gradient (5 cases). The abnormal single peak material failed to bind to DNA in 5 cases (of 5 studied), whereas peak I material from 5 cases (of 5 studied), showing normal peak I-peak II ratios, bound normally to DNA. In 3 cases (of 3 studied), the abnormal single peak material had an S value of 2–2.5S, whereas in 5 cases with normal peak I-peak II ratios, the S values were 3.5S and 8.5S, respectively. We hypothesize that those leukemias with abnormal binder characteristics cannot respond to glucocorticoid therapy.

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Characterization of glucocorticoid receptors in animal lymphoblastic disease: correlation with response to single-agent glucocorticoid treatment

Blood ◽

10.1182/blood.v63.2.380.380 ◽

1984 ◽

Vol 63 (2) ◽

pp. 380-383 ◽

Cited By ~ 8

Author(s):

R Bell ◽

S Cotter ◽

A Lillquist ◽

S Sallan ◽

R McCaffrey

Keyword(s):

Glucocorticoid Receptors ◽

Single Agent ◽

Complete Response ◽

Single Peak ◽

Human Leukemia ◽

Normal Pattern ◽

Glucocorticoid Treatment ◽

Normal Tissues ◽

Low Salt

Abstract The clinical significance of initial DEAE chromatography of glucocorticoid binders in lymphoblastic disease was evaluated in an animal model. Domestic cats and dogs with lymphoblastic disease were treated with prednisone, 2 mg/kg/day, for 14 days, and the outcome of therapy was correlated with DEAE chromatograms of glucocorticoid binders, using 3H-triamcinolone as ligand. Six of 30 animals had a single-peak low-salt binder species, similar to that seen in a subset of human leukemia, and none of these responded. Of the 29 animals with chromatograms identical to normal tissues, 6 had a complete response and another 11 a partial response. This distribution of responders is statistically significant (p = 0.02). Thus, the leukemia-associated single-peak DEAE species appears to be associated with glucocorticoid resistance, as defined by clinical responsiveness. In contrast, the two- peak normal pattern is a necessary, but insufficient, criterion for defining responsive disease.

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Biochemical and biophysical characterization of glucocorticoid receptors in normal lymphoid tissue

Blood ◽

10.1182/blood.v58.2.263.bloodjournal582263 ◽

1981 ◽

Vol 58 (2) ◽

pp. 263-267

Author(s):

R McCaffrey ◽

A Lillquist ◽

R Bell

Keyword(s):

Glucocorticoid Receptors ◽

Lymphoblastic Leukemia ◽

Gradient Centrifugation ◽

Lymphoid Cells ◽

Leukemia Cells ◽

Lymphoid Tissues ◽

Biophysical Characterization ◽

Two Component ◽

Heat Activation

3H-triamcinolone acetonide labeled glucocorticoid receptors in normal lymphoid tissues can be resolved into two component by DEAE chromatography: peak I elutes at 0.04 M salt and peak II is 0.22 M salt. By glycerol gradient centrifugation, peak I is 3.5S and peak II 8.5S. Peak I binds to DNA and chromatin, while peak II binds to neither. After heat activation, peak II alters its coefficient of sedimentation to 3.5S and on DEAE rechromatography changes its elution position to 0.04 M salt (peak I area) and acquires affinity for DNA. Glucocorticoid receptors in lymphoblastic leukemia cells can now be characterized using these techniques and compared to receptors in normal lymphoid cells.

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Characterization of glucocorticoid receptors in animal lymphoblastic disease: correlation with response to single-agent glucocorticoid treatment

Blood ◽

10.1182/blood.v63.2.380.bloodjournal632380 ◽

1984 ◽

Vol 63 (2) ◽

pp. 380-383

Author(s):

R Bell ◽

S Cotter ◽

A Lillquist ◽

S Sallan ◽

R McCaffrey

Keyword(s):

Glucocorticoid Receptors ◽

Single Agent ◽

Complete Response ◽

Single Peak ◽

Human Leukemia ◽

Normal Pattern ◽

Glucocorticoid Treatment ◽

Normal Tissues ◽

Low Salt

The clinical significance of initial DEAE chromatography of glucocorticoid binders in lymphoblastic disease was evaluated in an animal model. Domestic cats and dogs with lymphoblastic disease were treated with prednisone, 2 mg/kg/day, for 14 days, and the outcome of therapy was correlated with DEAE chromatograms of glucocorticoid binders, using 3H-triamcinolone as ligand. Six of 30 animals had a single-peak low-salt binder species, similar to that seen in a subset of human leukemia, and none of these responded. Of the 29 animals with chromatograms identical to normal tissues, 6 had a complete response and another 11 a partial response. This distribution of responders is statistically significant (p = 0.02). Thus, the leukemia-associated single-peak DEAE species appears to be associated with glucocorticoid resistance, as defined by clinical responsiveness. In contrast, the two- peak normal pattern is a necessary, but insufficient, criterion for defining responsive disease.

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Biochemical and biophysical characterization of glucocorticoid receptors in normal lymphoid tissue

Blood ◽

10.1182/blood.v58.2.263.263 ◽

1981 ◽

Vol 58 (2) ◽

pp. 263-267 ◽

Cited By ~ 2

Author(s):

R McCaffrey ◽

A Lillquist ◽

R Bell

Keyword(s):

Glucocorticoid Receptors ◽

Lymphoblastic Leukemia ◽

Gradient Centrifugation ◽

Lymphoid Cells ◽

Leukemia Cells ◽

Lymphoid Tissues ◽

Biophysical Characterization ◽

Two Component ◽

Heat Activation

Abstract 3H-triamcinolone acetonide labeled glucocorticoid receptors in normal lymphoid tissues can be resolved into two component by DEAE chromatography: peak I elutes at 0.04 M salt and peak II is 0.22 M salt. By glycerol gradient centrifugation, peak I is 3.5S and peak II 8.5S. Peak I binds to DNA and chromatin, while peak II binds to neither. After heat activation, peak II alters its coefficient of sedimentation to 3.5S and on DEAE rechromatography changes its elution position to 0.04 M salt (peak I area) and acquires affinity for DNA. Glucocorticoid receptors in lymphoblastic leukemia cells can now be characterized using these techniques and compared to receptors in normal lymphoid cells.

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Pre-Clinical Evaluation of the Proteasome Inhibitor Ixazomib against Bortezomib-Resistant Leukemia Cells and Primary Acute Leukemia Cells

Cells ◽

10.3390/cells10030665 ◽

2021 ◽

Vol 10 (3) ◽

pp. 665

Author(s):

Margot S.F. Roeten ◽

Johan van Meerloo ◽

Zinia J. Kwidama ◽

Giovanna ter Huizen ◽

Wouter H. Segerink ◽

...

Keyword(s):

Acute Leukemia ◽

Cell Lines ◽

Proteasome Inhibitor ◽

Ex Vivo ◽

Lymphoblastic Leukemia ◽

Unmet Need ◽

Leukemia Cell ◽

Leukemia Cells ◽

Cross Resistance ◽

Proteasome Subunit

At present, 20–30% of children with acute leukemia still relapse from current chemotherapy protocols, underscoring the unmet need for new treatment options, such as proteasome inhibition. Ixazomib (IXA) is an orally available proteasome inhibitor, with an improved safety profile compared to Bortezomib (BTZ). The mechanism of action (proteasome subunit inhibition, apoptosis induction) and growth inhibitory potential of IXA vs. BTZ were tested in vitro in human (BTZ-resistant) leukemia cell lines. Ex vivo activity of IXA vs. BTZ was analyzed in 15 acute lymphoblastic leukemia (ALL) and 9 acute myeloid leukemia (AML) primary pediatric patient samples. BTZ demonstrated more potent inhibitory effects on constitutive β5 and immunoproteasome β5i proteasome subunit activity; however, IXA more potently inhibited β1i subunit than BTZ (70% vs. 29% at 2.5 nM). In ALL/AML cell lines, IXA conveyed 50% growth inhibition at low nanomolar concentrations, but was ~10-fold less potent than BTZ. BTZ-resistant cells (150–160 fold) displayed similar (100-fold) cross-resistance to IXA. Finally, IXA and BTZ exhibited anti-leukemic effects for primary ex vivo ALL and AML cells; mean LC50 (nM) for IXA: 24 ± 11 and 30 ± 8, respectively, and mean LC50 for BTZ: 4.5 ± 1 and 11 ± 4, respectively. IXA has overlapping mechanisms of action with BTZ and showed anti-leukemic activity in primary leukemic cells, encouraging further pre-clinical in vivo evaluation.

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Alteration of hepatic glucocorticoid receptor stability and nuclear binding in vitro by citrate

Biochemical Journal ◽

10.1042/bj2100259 ◽

1983 ◽

Vol 210 (1) ◽

pp. 259-263 ◽

Cited By ~ 7

Author(s):

J Hubbard ◽

M Kalimi

Keyword(s):

Glucocorticoid Receptor ◽

Glucocorticoid Receptors ◽

Density Gradient Centrifugation ◽

Gradient Centrifugation ◽

Deae Cellulose ◽

Sucrose Density Gradient ◽

Sucrose Density Gradient Centrifugation ◽

Nuclear Binding ◽

Wide Range

Citrate greatly stabilized rat hepatic unbound glucocorticoid receptors in cell-free conditions at 4 degrees C with optimal effectiveness at 5-15 mM. Control receptors were inactivated at 4 degrees C with a half-life of less than 12 h. However, in the presence of 10 mM-citrate, unbound receptors were almost completely stabilized for 48 h at 4 degrees C. Citrate at a concentration of 1-2 mM yielded half-maximal stabilization. The stabilizing effect of citrate was rather specific, as succinate, alpha-oxoglutarate, oxaloacetate, malate and pyruvate had no apparent stabilizing action. Citrate stabilized receptors over a wide range of H+ concentrations, with complete protection between pH 6.5 and 8.5. In addition, citrate appeared to have a significant effect on glucocorticoid-receptor complex activation into a nuclear binding form. Thus 5-10 mM-citrate enhanced nuclear binding, with optimal activation achieved at 10 mM concentration. As analysed by sucrose-density-gradient centrifugation and DEAE-cellulose chromatography, no apparent change was observed in the physical characteristics of the glucocorticoid receptor in the presence of citrate.

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