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| Status |
Public on May 01, 2020 |
| Title |
Glucocorticoid Receptors are Required Effectors of TGFb1-Induced p38 MAPK Signaling to Advanced Cancer Phenotypes in Triple Negative Breast Cancer |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Altered signaling pathways typify breast cancer and serve as direct inputs to steroid hormone receptor sensors. We previously reported that phospho-Ser134-GR (pS134-GR) species are elevated in triple negative breast cancer (TNBC) and cooperate with hypoxia-inducible factors, providing a novel avenue for activation of GR in response to local or cellular stress. We probed GR regulation by factors (cytokines, growth factors) that are rich within the tumor microenvironment (TME). TNBC cells harboring endogenous wild-type (wt) or S134A-GR species were created by CRISPR/Cas knock-in and subjected to transwell migration, invasion, soft-agar colony formation, and tumorsphere assays. RNA-Seq was employed to identify pS134-GR target genes that are regulated both basally (intrinsic) or by TGFβ in the absence of exogenously added GR ligands. Regulation of selected basal and TGFβ-induced pS134-GR target genes was validated by qRT-PCR and chromatin immunoprecipitation assays. Bioinformatics tools were used to probe public data sets for expression of pS134-GR 24-gene signatures. In the absence of GR ligands, GR is transcriptionally activated via p38-dependent phosphorylation of Ser134 as a mechanism of homeostatic stress-sensing and regulated upon exposure of TNBC cells to TME-derived agents. The ligand-independent pS134-GR transcriptome encompasses TGFβ1 and MAPK signaling gene sets associated with TNBC cell survival and migration/invasion. Accordingly, pS134-GR was essential for TNBC cell anchorage-independent growth in soft-agar, migration, invasion, and tumorsphere formation, an in vitro readout of cancer stemness properties. Both pS134-GR and expression of the MAPK-scaffolding molecule 14-3-3ζ were essential for a functionally intact p38 MAPK signaling pathway downstream of MAP3K5/ASK1, indicative of a feed-forward signaling loop wherein self-perpetuated GR phosphorylation enables cancer cell autonomy. A 24-gene pS134-GR-dependent signature induced by TGFβ1 predicts shortened overall survival in breast cancer patients. Phospho-S134-GR is a critical downstream effector of p38 MAPK signaling and TNBC migration/invasion, survival, and stemness properties. Our studies define a ligand-independent role for GR as a homeostatic “sensor” of intrinsic stimuli as well as extrinsic factors rich within the TME (TGFβ1) that enables potent activation of the p38 MAPK stress-sensing pathway and nominate pS134-GR as a therapeutic target in aggressive TNBC.
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| Overall design |
Examination of the effect of TGFβ1 in TNBC cell line MDA-MB-231 with mutation in the glucocorticoid receptor on Ser134.
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| Contributor(s) |
Lange CA, Perez Kerkvliet C |
| Citation(s) |
32357907 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 CA192178 |
Inducible PTK6 expression drives oncogenic signaling in breast cancer |
UNIVERSITY OF MINNESOTA |
Carol A Lange |
| F30 CA228261 |
PhosphoSer134 GR, TGF-? and 14-3-3-zeta cooperate to promote progression of Triple Negative Breast Cancer |
UNIVERSITY OF MINNESOTA |
Carlos Jesus Perez Kerkvliet |
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| Submission date |
Apr 10, 2020 |
| Last update date |
Jul 31, 2020 |
| Contact name |
Carol Lange |
| E-mail(s) |
lange047@umn.edu
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| Phone |
7876780772
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| Organization name |
University of Minnesota
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| Department |
Medicine
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| Lab |
CCRB 3-240, B81, Lange
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| Street address |
2231 6th St SE
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| City |
Minneapolis |
| State/province |
Minnesota |
| ZIP/Postal code |
55455 |
| Country |
USA |
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| Platforms (1) |
| GPL16791 |
Illumina HiSeq 2500 (Homo sapiens) |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA624279 |
| SRA |
SRP255946 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE148444_RAW.tar |
2.3 Mb |
(http)(custom) |
TAR (of TXT) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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