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Increased HSD11β1 Expression in Human Leiomyomatous Uteri: Implication for Enhanced Glucocorticoid Signaling

ABSTRACT: Increased HSD11β1 Expression in Human Leiomyomatous Uteri: Implication for Enhanced Glucocorticoid Signaling

PROVIDER: PRJNA1238321 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR32775759_1.fastq.gz	Fastqsanger.gz
	SRR32775759_2.fastq.gz	Fastqsanger.gz
	SRR32775760_1.fastq.gz	Fastqsanger.gz
	SRR32775760_2.fastq.gz	Fastqsanger.gz
	SRR32775761_1.fastq.gz	Fastqsanger.gz

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Increased HSD11β1 Expression in Human Leiomyomatous Uteri: Implication for Enhanced Glucocorticoid Signaling

Project description:Abstract: FK506-binding-protein-51 (FKBP51) is a glucocorticoid-induced co-chaperone protein previously shown to bind glucocorticoid receptor (GR), inhibiting its transcriptional activity. We previously found increased FKBP51 levels in uterine leiomyoma versus paired myometrium. To test the hypothesis that elevated FKBP51 levels contribute to leiomyoma pathogenesis by altering GR signaling. Material Method: RNA-sequencing was performed in leiomyoma cell cultures transfected with scrambled or FKBP5-siRNA for 48-h, then treated with vehicle or dexamethasone (DEX) for 24-h. Differentially expressed genes, including HSD11B1, CNN1, and LAMA2 were analyzed by qPCR. Hydroxysteroid 11-beta dehydrogenase 1 (HSD11β1) expression was analyzed in leiomyoma, leiomyoma-adjacent paired myometrium, myometrium from patients without leiomyoma, and human endometrial stromal cells (HESC) by qPCR and immunohistochemistry in women with or without uterine leiomyoma. HSD11B1 mRNA and protein levels in leiomyoma, paired and normal myometrium, and HESC cells and tissue. Results: HSD11β1 expression was higher in paired myometrial and leiomyoma tissues versus normal myometrium (P<0.02). DEX treatment increased HSD11B1 transcription in normal myometrial and HESC cultures, but to a significantly greater extent in leiomyoma. However, FKBP5-silencing blunted DEX-induced HSD11B1 transcription. DEX-treatment reduced LAMA2 and increased CNN1 levels (coding for extracellular matrix and smooth muscle proteins, respectively) in FKBP5-silenced versus scrambled-siRNA leiomyoma cultures. Conclusions: FKBP51 not only inhibits but can augment GR-mediated transcription. Importantly, FKBP51-GR interactions increase HSD11B1 levels in leiomyoma cells, generating a pathological FKBP51-GR-HSD11β1 circle, altering transcription of downstream extracellular matrix and smooth muscle genes to induce a myofibroblast phenotype, thereby possibly contributing to leiomyoma pathogenesis.

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Cryptochromes mediate rhythmic repression of the glucocorticoid receptor

Project description:The circadian transcriptional repressors cryptochromes 1 (Cry1) and 2 (Cry2) interact with the C-terminus of the glucocorticoid receptor (GR) and are required for transrepression in response to the synthetic GR ligand dexamethasone (Dex) in mouse embryonic fibroblasts. Dex induction of many genes was increased in Cry-deficient fibroblasts suggesting that cryptochromes oppose transactivation in addition to contributing to transrepression. In mice, genetic loss of Cry1 and/or Cry2 resulted in glucose intolerance and constitutively high levels of circulating corticosterone, suggesting reduced glucocorticoid suppression of the hypothalamic-pituitary-adrenal axis coupled with increased sensitivity to the hyperglycemic effects of glucocorticoid-mediated transactivation in the liver. Cry1 and Cry2 association with a GRE in the Pck1 promoter was stimulated by Dex, and Dex-induced transcription of pck1 was strikingly increased in Cry-deficient livers. Finally, cry1-/-;cry2-/- mice subjected to 8 weeks of chronic Dex treatment exhibited incomplete suppression of circulating corticosterone and greater glucose intolerance compared with wildtype littermates subjected to the same chronic treatment, consistent with enhanced transcriptional response to the synthetic glucocorticoid ligand. Total RNA was obtained from WT and Cry1/2 KO MEFs treated with Dexamethasone (1uM) or control EtOH for 16 hours.

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Project description:The expression of K-ras transgene is induced in K-rasV12+/-/Cre+/+/ROSA26R-LacZ+/+ transgenic mice uteri, which were primed with hormone treatment. Decidualization was artificially induced in these uteri and hormone injection was withdrawn to induced menstruation-like endometrial degeneration. Microarray was then used to study the molecular changes in these menstruating uteri following the K-ras activation. Uterine tissues were collected from K-ras induced and littermate controled menstruating uteri. Experiments were carried out using Illumina Mouse WG-6 BeadChips (n=6 in each group). Data were normalized in R environment using the Lumi Bioconductor package.

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