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Transcriptional Pathways in cPGI2-Induced Adipocyte Progenitor Activation for Browning.

Bayindir I, Babaeikelishomi R, Kocanova S, Sousa IS, Lerch S, Hardt O, Wild S, Bosio A, Bystricky K, Herzig S, Vegiopoulos A - Front Endocrinol (Lausanne) (2015)

Bottom Line: We demonstrate the specific and broad induction of thermogenic gene expression by PGI2 signaling in the absence of lineage conversion, and reveal the previously unidentified nuclear relocalization of the Ucp1 gene locus in association with transcriptional activation.By profiling the time course of the progenitor response, we show that PGI2 signaling promoted progenitor cell activation through cell cycle and adhesion pathways prior to metabolic maturation toward an oxidative cell phenotype.Our results highlight the importance of core progenitor activation pathways for the recruitment of thermogenic cells and provide a resource for further mechanistic investigation.

View Article: PubMed Central - PubMed

Affiliation: DKFZ Junior Group Metabolism and Stem Cell Plasticity, German Cancer Research Center , Heidelberg , Germany.

ABSTRACT
De novo formation of beige/brite adipocytes from progenitor cells contributes to the thermogenic adaptation of adipose tissue and holds great potential for the therapeutic remodeling of fat as a treatment for obesity. Despite the recent identification of several factors regulating browning of white fat, there is a lack of physiological cell models for the mechanistic investigation of progenitor-mediated beige/brite differentiation. We have previously revealed prostacyclin (PGI2) as one of the few known endogenous extracellular mediators promoting de novo beige/brite formation by relaying β-adrenergic stimulation to the progenitor level. Here, we present a cell model based on murine primary progenitor cells defined by markers previously shown to be relevant for in vivo browning, including a simplified isolation procedure. We demonstrate the specific and broad induction of thermogenic gene expression by PGI2 signaling in the absence of lineage conversion, and reveal the previously unidentified nuclear relocalization of the Ucp1 gene locus in association with transcriptional activation. By profiling the time course of the progenitor response, we show that PGI2 signaling promoted progenitor cell activation through cell cycle and adhesion pathways prior to metabolic maturation toward an oxidative cell phenotype. Our results highlight the importance of core progenitor activation pathways for the recruitment of thermogenic cells and provide a resource for further mechanistic investigation.

No MeSH data available.


Related in: MedlinePlus

cPGI2 induces progenitor activation through cell cycle and adhesion pathways. Lin−CD29+CD34+Sca-1+ cells were cultured in adipogenic media ± cPGI2 for 24 h. (A–C) RNA was obtained for expression profiling with Illumina beadchip arrays (n = 3). Enrichment plots of the indicated gene sets were obtained by GSEA (cPGI2 vs. Control) with the KEGG pathway [(A,C), see Table 2] or the transcription factor motif (B) gene set collections. Vertical bars represent the individual genes of the gene set/pathway ranked according to their regulation by cPGI2. X-axis values represent the rank within the complete ranked gene list (transcriptome). The enrichment score (ES) reflects the degree to which a gene set is overrepresented at the top or bottom of the complete ranked gene list. (E,F) Cells were pulse-labeled with EdU for 1 h, processed for EdU-AlexaFluor647 and propidium iodide staining, and analyzed by flow cytometry (n = 3). Cells cultured without EdU served as negative control (D). The mean percentage of EdU+ cells is shown (* indicates t-test cPGI2 vs. Control p = 0.003).
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Figure 4: cPGI2 induces progenitor activation through cell cycle and adhesion pathways. Lin−CD29+CD34+Sca-1+ cells were cultured in adipogenic media ± cPGI2 for 24 h. (A–C) RNA was obtained for expression profiling with Illumina beadchip arrays (n = 3). Enrichment plots of the indicated gene sets were obtained by GSEA (cPGI2 vs. Control) with the KEGG pathway [(A,C), see Table 2] or the transcription factor motif (B) gene set collections. Vertical bars represent the individual genes of the gene set/pathway ranked according to their regulation by cPGI2. X-axis values represent the rank within the complete ranked gene list (transcriptome). The enrichment score (ES) reflects the degree to which a gene set is overrepresented at the top or bottom of the complete ranked gene list. (E,F) Cells were pulse-labeled with EdU for 1 h, processed for EdU-AlexaFluor647 and propidium iodide staining, and analyzed by flow cytometry (n = 3). Cells cultured without EdU served as negative control (D). The mean percentage of EdU+ cells is shown (* indicates t-test cPGI2 vs. Control p = 0.003).

Mentions: We next sought to determine the transcriptional pathways underlying the early progenitor response to cPGI2. GSEA at 24 h of cPGI2 treatment using the KEGG pathway collection revealed the marked enrichment of cell cycle and proliferation pathways in the gene fraction up-regulated by cPGI2 (Table 2; Figure 4A).


Transcriptional Pathways in cPGI2-Induced Adipocyte Progenitor Activation for Browning.

Bayindir I, Babaeikelishomi R, Kocanova S, Sousa IS, Lerch S, Hardt O, Wild S, Bosio A, Bystricky K, Herzig S, Vegiopoulos A - Front Endocrinol (Lausanne) (2015)

cPGI2 induces progenitor activation through cell cycle and adhesion pathways. Lin−CD29+CD34+Sca-1+ cells were cultured in adipogenic media ± cPGI2 for 24 h. (A–C) RNA was obtained for expression profiling with Illumina beadchip arrays (n = 3). Enrichment plots of the indicated gene sets were obtained by GSEA (cPGI2 vs. Control) with the KEGG pathway [(A,C), see Table 2] or the transcription factor motif (B) gene set collections. Vertical bars represent the individual genes of the gene set/pathway ranked according to their regulation by cPGI2. X-axis values represent the rank within the complete ranked gene list (transcriptome). The enrichment score (ES) reflects the degree to which a gene set is overrepresented at the top or bottom of the complete ranked gene list. (E,F) Cells were pulse-labeled with EdU for 1 h, processed for EdU-AlexaFluor647 and propidium iodide staining, and analyzed by flow cytometry (n = 3). Cells cultured without EdU served as negative control (D). The mean percentage of EdU+ cells is shown (* indicates t-test cPGI2 vs. Control p = 0.003).
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Figure 4: cPGI2 induces progenitor activation through cell cycle and adhesion pathways. Lin−CD29+CD34+Sca-1+ cells were cultured in adipogenic media ± cPGI2 for 24 h. (A–C) RNA was obtained for expression profiling with Illumina beadchip arrays (n = 3). Enrichment plots of the indicated gene sets were obtained by GSEA (cPGI2 vs. Control) with the KEGG pathway [(A,C), see Table 2] or the transcription factor motif (B) gene set collections. Vertical bars represent the individual genes of the gene set/pathway ranked according to their regulation by cPGI2. X-axis values represent the rank within the complete ranked gene list (transcriptome). The enrichment score (ES) reflects the degree to which a gene set is overrepresented at the top or bottom of the complete ranked gene list. (E,F) Cells were pulse-labeled with EdU for 1 h, processed for EdU-AlexaFluor647 and propidium iodide staining, and analyzed by flow cytometry (n = 3). Cells cultured without EdU served as negative control (D). The mean percentage of EdU+ cells is shown (* indicates t-test cPGI2 vs. Control p = 0.003).
Mentions: We next sought to determine the transcriptional pathways underlying the early progenitor response to cPGI2. GSEA at 24 h of cPGI2 treatment using the KEGG pathway collection revealed the marked enrichment of cell cycle and proliferation pathways in the gene fraction up-regulated by cPGI2 (Table 2; Figure 4A).

Bottom Line: We demonstrate the specific and broad induction of thermogenic gene expression by PGI2 signaling in the absence of lineage conversion, and reveal the previously unidentified nuclear relocalization of the Ucp1 gene locus in association with transcriptional activation.By profiling the time course of the progenitor response, we show that PGI2 signaling promoted progenitor cell activation through cell cycle and adhesion pathways prior to metabolic maturation toward an oxidative cell phenotype.Our results highlight the importance of core progenitor activation pathways for the recruitment of thermogenic cells and provide a resource for further mechanistic investigation.

View Article: PubMed Central - PubMed

Affiliation: DKFZ Junior Group Metabolism and Stem Cell Plasticity, German Cancer Research Center , Heidelberg , Germany.

ABSTRACT
De novo formation of beige/brite adipocytes from progenitor cells contributes to the thermogenic adaptation of adipose tissue and holds great potential for the therapeutic remodeling of fat as a treatment for obesity. Despite the recent identification of several factors regulating browning of white fat, there is a lack of physiological cell models for the mechanistic investigation of progenitor-mediated beige/brite differentiation. We have previously revealed prostacyclin (PGI2) as one of the few known endogenous extracellular mediators promoting de novo beige/brite formation by relaying β-adrenergic stimulation to the progenitor level. Here, we present a cell model based on murine primary progenitor cells defined by markers previously shown to be relevant for in vivo browning, including a simplified isolation procedure. We demonstrate the specific and broad induction of thermogenic gene expression by PGI2 signaling in the absence of lineage conversion, and reveal the previously unidentified nuclear relocalization of the Ucp1 gene locus in association with transcriptional activation. By profiling the time course of the progenitor response, we show that PGI2 signaling promoted progenitor cell activation through cell cycle and adhesion pathways prior to metabolic maturation toward an oxidative cell phenotype. Our results highlight the importance of core progenitor activation pathways for the recruitment of thermogenic cells and provide a resource for further mechanistic investigation.

No MeSH data available.


Related in: MedlinePlus