The long-term objectives of this project focus on the human glucocorticoid receptor (GR), its gene, and glucocorticoid action in human leukemias. DNA sequences encoding a large part of the human GR have just been obtained, in what may be the first isolation of a gene encoding a human transcription regulatory protein, one whose occupancy by steroids leads to death of leukemic lymphoblasts. This DNA, along with clones of sensitive and resistant cells from the well-studied acute lymphoblastic leukemic line (CEM) will be used in a combination of methods, involving molecular cloning, protein purification and microsequencing, steroid binding, and somatic cell genetics to study human GR and its mechanism of action.
Specific aims will include comparisons of human, rat and mouse GR genes, measurement of GR mRNA levels and comparison of GR genes in mutant and wild-type CEM cells as well as in genetically glucocorticoid-resistant humans and monkeys. The human GR will be further purified so that sequence data and monoclonal antibodies may be obtained. The binding of phenylpyrazolo glucocorticoids will be studied to see whether they interact solely with the classic GR. Cell ploidy, exposure time to and concentration of glucocorticoids will be evaluated for their bearing on effective use of the steroids to produce cell kill. Additional r+ly- (lysis defective) cell lines of several classes of leukemia will be treated with azadeoxycytidine to see if they will shift to ly+. Somatic cell hybrids between r+ly- cells and resistant cells with defective receptors will be made to test the function of the receptor from the ly- cell. Steroid sensitive CEM cells will be examined for oncogene/transforming gene expression and phosphatidyl inositol turnover, in the presence and absence of glucocorticoids.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA041407-03
Application #
3181854
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1985-12-01
Project End
1990-11-30
Budget Start
1987-12-01
Budget End
1988-11-30
Support Year
3
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Texas Medical Br Galveston
Department
Type
Schools of Medicine
DUNS #
041367053
City
Galveston
State
TX
Country
United States
Zip Code
77555
Webb, M S; Miller, A L; Howard, T L et al. (2018) Sequential gene regulatory events leading to glucocorticoid-evoked apoptosis of CEM human leukemic cells:interactions of MAPK, MYC and glucocorticoid pathways. Mol Cell Endocrinol 471:118-130
Aiyar, Sarah E; Park, Hoyong; Aldo, Paulomi B et al. (2010) TMS, a chemically modified herbal derivative of resveratrol, induces cell death by targeting Bax. Breast Cancer Res Treat 124:265-77
Garza, Anna S; Miller, Aaron L; Johnson, Betty H et al. (2009) Converting cell lines representing hematological malignancies from glucocorticoid-resistant to glucocorticoid-sensitive: signaling pathway interactions. Leuk Res 33:717-27
Ji, Zhenyu; Mei, Fang C; Miller, Aaron L et al. (2008) Protein kinase A (PKA) isoform RIIbeta mediates the synergistic killing effect of cAMP and glucocorticoid in acute lymphoblastic leukemia cells. J Biol Chem 283:21920-5
Webb, M S; Miller, A L; Thompson, E Brad (2007) In CEM cells the autosomal deafness gene dfna5 is regulated by glucocorticoids and forskolin. J Steroid Biochem Mol Biol 107:15-21
Miller, Aaron L; Webb, M Scott; Thompson, E Brad (2007) Comparison of two structurally diverse glucocorticoid receptor agonists: cortivazol selectively regulates a distinct set of genes separate from dexamethasone in CEM cells. Steroids 72:673-81
Ji, Zhenyu; Mei, Fang C; Johnson, Betty H et al. (2007) Protein kinase A, not Epac, suppresses hedgehog activity and regulates glucocorticoid sensitivity in acute lymphoblastic leukemia cells. J Biol Chem 282:37370-7
Ilies, Marc Antoniu; Seitz, William A; Johnson, Betty H et al. (2006) Lipophilic pyrylium salts in the synthesis of efficient pyridinium-based cationic lipids, gemini surfactants, and lipophilic oligomers for gene delivery. J Med Chem 49:3872-87
Copik, Alicja J; Webb, M Scott; Miller, Aaron L et al. (2006) Activation function 1 of glucocorticoid receptor binds TATA-binding protein in vitro and in vivo. Mol Endocrinol 20:1218-30
Miller, Aaron L; Webb, M Scott; Copik, Alicja J et al. (2005) p38 Mitogen-activated protein kinase (MAPK) is a key mediator in glucocorticoid-induced apoptosis of lymphoid cells: correlation between p38 MAPK activation and site-specific phosphorylation of the human glucocorticoid receptor at serine 211. Mol Endocrinol 19:1569-83

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