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Retinoic Acid Mediates Visceral-Specific Adipogenic Defects of Human Adipose-Derived Stem Cells

View Article: PubMed Central - PubMed

ABSTRACT

Increased visceral fat, rather than subcutaneous fat, during the onset of obesity is associated with a higher risk of developing metabolic diseases. The inherent adipogenic properties of human adipose-derived stem cells (ASCs) from visceral depots are compromised compared with those of ASCs from subcutaneous depots, but little is known about the underlying mechanisms. Using ontological analysis of global gene expression studies, we demonstrate that many genes involved in retinoic acid (RA) synthesis or regulated by RA are differentially expressed in human tissues and ASCs from subcutaneous and visceral fat. The endogenous level of RA is higher in visceral ASCs; this is associated with upregulation of the RA synthesis gene through the visceral-specific developmental factor WT1. Excessive RA-mediated activity impedes the adipogenic capability of ASCs at early but not late stages of adipogenesis, which can be reversed by antagonism of RA receptors or knockdown of WT1. Our results reveal the developmental origin of adipocytic properties and the pathophysiological contributions of visceral fat depots.

No MeSH data available.


RA-mediated adipogenic defects are reversed by the antagonism of the downstream target RAR. A: A representative graph showing the relative fluorescence units (RFUs) of AdipoRed staining on VS ASCs from S11. Adipocyte differentiation was induced in ASCs using a standard adipogenic cocktail with or without pretreatment/treatment with BMS493 and/or RA at various time points. The ASCs were treated with 1 μmol/L of BMS493 and/or 10 μmol/L of RA at the time points indicated: D0 (D−2 to D0), D3 (D−2 to D3), and D12 (D−2 to D12). *P < 0.05 and ^P < 0.05 denote significant fold change in RFUs corresponding to the quantitation of lipid accumulation during adipocyte differentiation (n = 2). B: Representative images (original magnification ×10) showing lipid accumulation (AdipoRed, green) in VS ASCs from S11 treated with 1 μmol/L of BMS493 and/or 10 μmol/L of RA at the different time points indicated: D0 (D−2 to D0), D3 (D−2 to D3), and D12 (D−2 to D12). For the purpose of presentation, the fluorescent intensities were enhanced to the same degree for all images. Scale bar = 100 µm (n = 2). C: A representative graph showing the RFUs of AdipoRed staining on VS ASCs from S11. ASCs were treated with LE135 (10 or 25 nmol/L) for 3 days upon the induction of adipogenic differentiation with a standard adipogenic cocktail. *P < 0.05 and **P < 0.01 denote significant fold change in RFUs (n = 2). D: Representative images (original magnification ×10) showing lipid accumulation (AdipoRed, green) in VS ASCs from S11 treated with LE135 (10 or 25 nmol/L) for 3 days upon the induction of adipogenic differentiation. For the purpose of presentation, the fluorescent intensities were enhanced to the same degree for all images. Scale bar = 100 µm (n = 2). Similar results were obtained from experiments using cells from S12.
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Figure 8: RA-mediated adipogenic defects are reversed by the antagonism of the downstream target RAR. A: A representative graph showing the relative fluorescence units (RFUs) of AdipoRed staining on VS ASCs from S11. Adipocyte differentiation was induced in ASCs using a standard adipogenic cocktail with or without pretreatment/treatment with BMS493 and/or RA at various time points. The ASCs were treated with 1 μmol/L of BMS493 and/or 10 μmol/L of RA at the time points indicated: D0 (D−2 to D0), D3 (D−2 to D3), and D12 (D−2 to D12). *P < 0.05 and ^P < 0.05 denote significant fold change in RFUs corresponding to the quantitation of lipid accumulation during adipocyte differentiation (n = 2). B: Representative images (original magnification ×10) showing lipid accumulation (AdipoRed, green) in VS ASCs from S11 treated with 1 μmol/L of BMS493 and/or 10 μmol/L of RA at the different time points indicated: D0 (D−2 to D0), D3 (D−2 to D3), and D12 (D−2 to D12). For the purpose of presentation, the fluorescent intensities were enhanced to the same degree for all images. Scale bar = 100 µm (n = 2). C: A representative graph showing the RFUs of AdipoRed staining on VS ASCs from S11. ASCs were treated with LE135 (10 or 25 nmol/L) for 3 days upon the induction of adipogenic differentiation with a standard adipogenic cocktail. *P < 0.05 and **P < 0.01 denote significant fold change in RFUs (n = 2). D: Representative images (original magnification ×10) showing lipid accumulation (AdipoRed, green) in VS ASCs from S11 treated with LE135 (10 or 25 nmol/L) for 3 days upon the induction of adipogenic differentiation. For the purpose of presentation, the fluorescent intensities were enhanced to the same degree for all images. Scale bar = 100 µm (n = 2). Similar results were obtained from experiments using cells from S12.

Mentions: Finally, to address whether adipogenic defects caused by excessive RA can be reversed by modulating the RA pathway in VS ASCs, we used BMS493, an antagonist of RARs (α, β, γ), at different points during adipocyte differentiation. The result of AdipoRed staining and quantification demonstrated that treatment of BMS493 during D−2 and D12 significantly improved the adipogenic capacity of VS ASCs (Fig. 8A and B). BMS493 also reversed the adipogenic defect caused by excessive RA at all the times tested (Fig. 8A and B). We also treated VS ASCs with LE135, a RARβ-specific antagonist. AdipoRed staining revealed that treatment with LE135 significantly improved the adipogenic potential of VS ASCs, as shown in Fig. 8C and D. Together, these results propose a potential therapeutic strategy of RAR or RARβ antagonism that can relieve adipogenic dysfunction mediated by excessive RA signaling in VS fat.


Retinoic Acid Mediates Visceral-Specific Adipogenic Defects of Human Adipose-Derived Stem Cells
RA-mediated adipogenic defects are reversed by the antagonism of the downstream target RAR. A: A representative graph showing the relative fluorescence units (RFUs) of AdipoRed staining on VS ASCs from S11. Adipocyte differentiation was induced in ASCs using a standard adipogenic cocktail with or without pretreatment/treatment with BMS493 and/or RA at various time points. The ASCs were treated with 1 μmol/L of BMS493 and/or 10 μmol/L of RA at the time points indicated: D0 (D−2 to D0), D3 (D−2 to D3), and D12 (D−2 to D12). *P < 0.05 and ^P < 0.05 denote significant fold change in RFUs corresponding to the quantitation of lipid accumulation during adipocyte differentiation (n = 2). B: Representative images (original magnification ×10) showing lipid accumulation (AdipoRed, green) in VS ASCs from S11 treated with 1 μmol/L of BMS493 and/or 10 μmol/L of RA at the different time points indicated: D0 (D−2 to D0), D3 (D−2 to D3), and D12 (D−2 to D12). For the purpose of presentation, the fluorescent intensities were enhanced to the same degree for all images. Scale bar = 100 µm (n = 2). C: A representative graph showing the RFUs of AdipoRed staining on VS ASCs from S11. ASCs were treated with LE135 (10 or 25 nmol/L) for 3 days upon the induction of adipogenic differentiation with a standard adipogenic cocktail. *P < 0.05 and **P < 0.01 denote significant fold change in RFUs (n = 2). D: Representative images (original magnification ×10) showing lipid accumulation (AdipoRed, green) in VS ASCs from S11 treated with LE135 (10 or 25 nmol/L) for 3 days upon the induction of adipogenic differentiation. For the purpose of presentation, the fluorescent intensities were enhanced to the same degree for all images. Scale bar = 100 µm (n = 2). Similar results were obtained from experiments using cells from S12.
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Related In: Results  -  Collection

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Figure 8: RA-mediated adipogenic defects are reversed by the antagonism of the downstream target RAR. A: A representative graph showing the relative fluorescence units (RFUs) of AdipoRed staining on VS ASCs from S11. Adipocyte differentiation was induced in ASCs using a standard adipogenic cocktail with or without pretreatment/treatment with BMS493 and/or RA at various time points. The ASCs were treated with 1 μmol/L of BMS493 and/or 10 μmol/L of RA at the time points indicated: D0 (D−2 to D0), D3 (D−2 to D3), and D12 (D−2 to D12). *P < 0.05 and ^P < 0.05 denote significant fold change in RFUs corresponding to the quantitation of lipid accumulation during adipocyte differentiation (n = 2). B: Representative images (original magnification ×10) showing lipid accumulation (AdipoRed, green) in VS ASCs from S11 treated with 1 μmol/L of BMS493 and/or 10 μmol/L of RA at the different time points indicated: D0 (D−2 to D0), D3 (D−2 to D3), and D12 (D−2 to D12). For the purpose of presentation, the fluorescent intensities were enhanced to the same degree for all images. Scale bar = 100 µm (n = 2). C: A representative graph showing the RFUs of AdipoRed staining on VS ASCs from S11. ASCs were treated with LE135 (10 or 25 nmol/L) for 3 days upon the induction of adipogenic differentiation with a standard adipogenic cocktail. *P < 0.05 and **P < 0.01 denote significant fold change in RFUs (n = 2). D: Representative images (original magnification ×10) showing lipid accumulation (AdipoRed, green) in VS ASCs from S11 treated with LE135 (10 or 25 nmol/L) for 3 days upon the induction of adipogenic differentiation. For the purpose of presentation, the fluorescent intensities were enhanced to the same degree for all images. Scale bar = 100 µm (n = 2). Similar results were obtained from experiments using cells from S12.
Mentions: Finally, to address whether adipogenic defects caused by excessive RA can be reversed by modulating the RA pathway in VS ASCs, we used BMS493, an antagonist of RARs (α, β, γ), at different points during adipocyte differentiation. The result of AdipoRed staining and quantification demonstrated that treatment of BMS493 during D−2 and D12 significantly improved the adipogenic capacity of VS ASCs (Fig. 8A and B). BMS493 also reversed the adipogenic defect caused by excessive RA at all the times tested (Fig. 8A and B). We also treated VS ASCs with LE135, a RARβ-specific antagonist. AdipoRed staining revealed that treatment with LE135 significantly improved the adipogenic potential of VS ASCs, as shown in Fig. 8C and D. Together, these results propose a potential therapeutic strategy of RAR or RARβ antagonism that can relieve adipogenic dysfunction mediated by excessive RA signaling in VS fat.

View Article: PubMed Central - PubMed

ABSTRACT

Increased visceral fat, rather than subcutaneous fat, during the onset of obesity is associated with a higher risk of developing metabolic diseases. The inherent adipogenic properties of human adipose-derived stem cells (ASCs) from visceral depots are compromised compared with those of ASCs from subcutaneous depots, but little is known about the underlying mechanisms. Using ontological analysis of global gene expression studies, we demonstrate that many genes involved in retinoic acid (RA) synthesis or regulated by RA are differentially expressed in human tissues and ASCs from subcutaneous and visceral fat. The endogenous level of RA is higher in visceral ASCs; this is associated with upregulation of the RA synthesis gene through the visceral-specific developmental factor WT1. Excessive RA-mediated activity impedes the adipogenic capability of ASCs at early but not late stages of adipogenesis, which can be reversed by antagonism of RA receptors or knockdown of WT1. Our results reveal the developmental origin of adipocytic properties and the pathophysiological contributions of visceral fat depots.

No MeSH data available.