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FAD-dependent lysine-specific demethylase-1 regulates cellular energy expenditure.

Hino S, Sakamoto A, Nagaoka K, Anan K, Wang Y, Mimasu S, Umehara T, Yokoyama S, Kosai K, Nakao M - Nat Commun (2012)

Bottom Line: The lysine-specific demethylase-1 (LSD1) is a unique nuclear protein that utilizes flavin adenosine dinucleotide (FAD) as a cofactor.We find that the loss of LSD1 function, either by short interfering RNA or by selective inhibitors in adipocytes, induces a number of regulators of energy expenditure and mitochondrial metabolism such as PPARγ coactivator-1α resulting in the activation of mitochondrial respiration.In the adipose tissues from mice on a high-fat diet, expression of LSD1-target genes is reduced, compared with that in tissues from mice on a normal diet, which can be reverted by suppressing LSD1 function.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Cell Biology, Institute of Molecular Embryology and Genetics, the Global Center of Excellence 'Cell Fate Regulation Research and Education Unit', Kumamoto University, 860-0811, Japan. s-hino@kumamoto-u.ac.jp

ABSTRACT
Environmental factors such as nutritional state may act on the epigenome that consequently contributes to the metabolic adaptation of cells and the organisms. The lysine-specific demethylase-1 (LSD1) is a unique nuclear protein that utilizes flavin adenosine dinucleotide (FAD) as a cofactor. Here we show that LSD1 epigenetically regulates energy-expenditure genes in adipocytes depending on the cellular FAD availability. We find that the loss of LSD1 function, either by short interfering RNA or by selective inhibitors in adipocytes, induces a number of regulators of energy expenditure and mitochondrial metabolism such as PPARγ coactivator-1α resulting in the activation of mitochondrial respiration. In the adipose tissues from mice on a high-fat diet, expression of LSD1-target genes is reduced, compared with that in tissues from mice on a normal diet, which can be reverted by suppressing LSD1 function. Our data suggest a novel mechanism where LSD1 regulates cellular energy balance through coupling with cellular FAD biosynthesis.

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Inhibition of LSD1 induces energy-expenditure genes in obese adipose tissues.(a,b) Expression of LSD1 and BHC80 in epididymal WAT from ND (black bars) and HFD- (white bars) fed mice. After a 16-hour fasting period, epididymal WAT was collected, and the total RNA was used for quantitative RT–PCR (a). The expression level of the 36B4 gene was used as the internal control. Protein levels are shown by western blot analysis (b). (c) Expression of LSD1 target genes in ND- (black bars) and HFD- (white bars) fed mice. Values are means±s.d. of four mice, and are shown as fold changes relative to ND-fed mice. **P<0.01 versus ND-fed mice by Student's t-test. (d) Efficient introduction of adenovirus vector into cultured WAT. Adenovirus vector, Ad-CAG-eGFP was introduced into isolated epididymal WAT, and the eGFP expression was analysed by fluorescence microscopy. Scale bar indicates 50 μm. (e,f) Expression of energy-expenditure genes after adenovirus-mediated knockdown of LSD1 gene in cultured WAT ex vivo. Epididymal WAT was dissected from either HFD- (e) or ND-fed mice (f), followed by the infection of adenoviruses, Ad-shLSD1 (red bars) or Ad-sh control (black bars). (g) Efficient introduction of adenoviral vectors into epididymal WAT in vivo. Adenovirus vectors carrying the eGFP gene (Ad-CAG-eGFP) was injected into epididymal WAT after incising the outer coat of mice. Four days later, tissues were isolated for the microscopic analysis. Scale bar indicates 2 mm. (h) Expression of energy-expenditure genes after adenovirus-mediated reduction of LSD1 in epididymal WAT in vivo. Control (black bars) or LSD1 (red bars) shRNA-carrying adenoviruses were directly injected into epididymal WAT of HFD-fed mice 4 days before tissue isolation. Values are means±s.d. of triplicate samples. *P<0.05, **P<0.01 versus control shRNA by Student's t-test.
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f7: Inhibition of LSD1 induces energy-expenditure genes in obese adipose tissues.(a,b) Expression of LSD1 and BHC80 in epididymal WAT from ND (black bars) and HFD- (white bars) fed mice. After a 16-hour fasting period, epididymal WAT was collected, and the total RNA was used for quantitative RT–PCR (a). The expression level of the 36B4 gene was used as the internal control. Protein levels are shown by western blot analysis (b). (c) Expression of LSD1 target genes in ND- (black bars) and HFD- (white bars) fed mice. Values are means±s.d. of four mice, and are shown as fold changes relative to ND-fed mice. **P<0.01 versus ND-fed mice by Student's t-test. (d) Efficient introduction of adenovirus vector into cultured WAT. Adenovirus vector, Ad-CAG-eGFP was introduced into isolated epididymal WAT, and the eGFP expression was analysed by fluorescence microscopy. Scale bar indicates 50 μm. (e,f) Expression of energy-expenditure genes after adenovirus-mediated knockdown of LSD1 gene in cultured WAT ex vivo. Epididymal WAT was dissected from either HFD- (e) or ND-fed mice (f), followed by the infection of adenoviruses, Ad-shLSD1 (red bars) or Ad-sh control (black bars). (g) Efficient introduction of adenoviral vectors into epididymal WAT in vivo. Adenovirus vectors carrying the eGFP gene (Ad-CAG-eGFP) was injected into epididymal WAT after incising the outer coat of mice. Four days later, tissues were isolated for the microscopic analysis. Scale bar indicates 2 mm. (h) Expression of energy-expenditure genes after adenovirus-mediated reduction of LSD1 in epididymal WAT in vivo. Control (black bars) or LSD1 (red bars) shRNA-carrying adenoviruses were directly injected into epididymal WAT of HFD-fed mice 4 days before tissue isolation. Values are means±s.d. of triplicate samples. *P<0.05, **P<0.01 versus control shRNA by Student's t-test.

Mentions: To address the LSD1-dependent metabolic gene regulation in vivo, we performed gene-expression analyses in normal and obese adipose tissues of mice. Seven-week old C57BL/6J mice were fed a HFD for six weeks and acquired an obese state. Interestingly, in the adipose tissues of HFD-fed mice, the expression of LSD1 and BHC80 was markedly elevated compared with ND controls (Fig. 7a,b). In addition, the expression of the LSD1 target genes such as PGC-1α, PDK4 and FATP1 was significantly reduced in the obese WAT tissues (Fig. 7c), indicating an inverse correlation with LSD1/BHC80 expression.


FAD-dependent lysine-specific demethylase-1 regulates cellular energy expenditure.

Hino S, Sakamoto A, Nagaoka K, Anan K, Wang Y, Mimasu S, Umehara T, Yokoyama S, Kosai K, Nakao M - Nat Commun (2012)

Inhibition of LSD1 induces energy-expenditure genes in obese adipose tissues.(a,b) Expression of LSD1 and BHC80 in epididymal WAT from ND (black bars) and HFD- (white bars) fed mice. After a 16-hour fasting period, epididymal WAT was collected, and the total RNA was used for quantitative RT–PCR (a). The expression level of the 36B4 gene was used as the internal control. Protein levels are shown by western blot analysis (b). (c) Expression of LSD1 target genes in ND- (black bars) and HFD- (white bars) fed mice. Values are means±s.d. of four mice, and are shown as fold changes relative to ND-fed mice. **P<0.01 versus ND-fed mice by Student's t-test. (d) Efficient introduction of adenovirus vector into cultured WAT. Adenovirus vector, Ad-CAG-eGFP was introduced into isolated epididymal WAT, and the eGFP expression was analysed by fluorescence microscopy. Scale bar indicates 50 μm. (e,f) Expression of energy-expenditure genes after adenovirus-mediated knockdown of LSD1 gene in cultured WAT ex vivo. Epididymal WAT was dissected from either HFD- (e) or ND-fed mice (f), followed by the infection of adenoviruses, Ad-shLSD1 (red bars) or Ad-sh control (black bars). (g) Efficient introduction of adenoviral vectors into epididymal WAT in vivo. Adenovirus vectors carrying the eGFP gene (Ad-CAG-eGFP) was injected into epididymal WAT after incising the outer coat of mice. Four days later, tissues were isolated for the microscopic analysis. Scale bar indicates 2 mm. (h) Expression of energy-expenditure genes after adenovirus-mediated reduction of LSD1 in epididymal WAT in vivo. Control (black bars) or LSD1 (red bars) shRNA-carrying adenoviruses were directly injected into epididymal WAT of HFD-fed mice 4 days before tissue isolation. Values are means±s.d. of triplicate samples. *P<0.05, **P<0.01 versus control shRNA by Student's t-test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3316891&req=5

f7: Inhibition of LSD1 induces energy-expenditure genes in obese adipose tissues.(a,b) Expression of LSD1 and BHC80 in epididymal WAT from ND (black bars) and HFD- (white bars) fed mice. After a 16-hour fasting period, epididymal WAT was collected, and the total RNA was used for quantitative RT–PCR (a). The expression level of the 36B4 gene was used as the internal control. Protein levels are shown by western blot analysis (b). (c) Expression of LSD1 target genes in ND- (black bars) and HFD- (white bars) fed mice. Values are means±s.d. of four mice, and are shown as fold changes relative to ND-fed mice. **P<0.01 versus ND-fed mice by Student's t-test. (d) Efficient introduction of adenovirus vector into cultured WAT. Adenovirus vector, Ad-CAG-eGFP was introduced into isolated epididymal WAT, and the eGFP expression was analysed by fluorescence microscopy. Scale bar indicates 50 μm. (e,f) Expression of energy-expenditure genes after adenovirus-mediated knockdown of LSD1 gene in cultured WAT ex vivo. Epididymal WAT was dissected from either HFD- (e) or ND-fed mice (f), followed by the infection of adenoviruses, Ad-shLSD1 (red bars) or Ad-sh control (black bars). (g) Efficient introduction of adenoviral vectors into epididymal WAT in vivo. Adenovirus vectors carrying the eGFP gene (Ad-CAG-eGFP) was injected into epididymal WAT after incising the outer coat of mice. Four days later, tissues were isolated for the microscopic analysis. Scale bar indicates 2 mm. (h) Expression of energy-expenditure genes after adenovirus-mediated reduction of LSD1 in epididymal WAT in vivo. Control (black bars) or LSD1 (red bars) shRNA-carrying adenoviruses were directly injected into epididymal WAT of HFD-fed mice 4 days before tissue isolation. Values are means±s.d. of triplicate samples. *P<0.05, **P<0.01 versus control shRNA by Student's t-test.
Mentions: To address the LSD1-dependent metabolic gene regulation in vivo, we performed gene-expression analyses in normal and obese adipose tissues of mice. Seven-week old C57BL/6J mice were fed a HFD for six weeks and acquired an obese state. Interestingly, in the adipose tissues of HFD-fed mice, the expression of LSD1 and BHC80 was markedly elevated compared with ND controls (Fig. 7a,b). In addition, the expression of the LSD1 target genes such as PGC-1α, PDK4 and FATP1 was significantly reduced in the obese WAT tissues (Fig. 7c), indicating an inverse correlation with LSD1/BHC80 expression.

Bottom Line: The lysine-specific demethylase-1 (LSD1) is a unique nuclear protein that utilizes flavin adenosine dinucleotide (FAD) as a cofactor.We find that the loss of LSD1 function, either by short interfering RNA or by selective inhibitors in adipocytes, induces a number of regulators of energy expenditure and mitochondrial metabolism such as PPARγ coactivator-1α resulting in the activation of mitochondrial respiration.In the adipose tissues from mice on a high-fat diet, expression of LSD1-target genes is reduced, compared with that in tissues from mice on a normal diet, which can be reverted by suppressing LSD1 function.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Cell Biology, Institute of Molecular Embryology and Genetics, the Global Center of Excellence 'Cell Fate Regulation Research and Education Unit', Kumamoto University, 860-0811, Japan. s-hino@kumamoto-u.ac.jp

ABSTRACT
Environmental factors such as nutritional state may act on the epigenome that consequently contributes to the metabolic adaptation of cells and the organisms. The lysine-specific demethylase-1 (LSD1) is a unique nuclear protein that utilizes flavin adenosine dinucleotide (FAD) as a cofactor. Here we show that LSD1 epigenetically regulates energy-expenditure genes in adipocytes depending on the cellular FAD availability. We find that the loss of LSD1 function, either by short interfering RNA or by selective inhibitors in adipocytes, induces a number of regulators of energy expenditure and mitochondrial metabolism such as PPARγ coactivator-1α resulting in the activation of mitochondrial respiration. In the adipose tissues from mice on a high-fat diet, expression of LSD1-target genes is reduced, compared with that in tissues from mice on a normal diet, which can be reverted by suppressing LSD1 function. Our data suggest a novel mechanism where LSD1 regulates cellular energy balance through coupling with cellular FAD biosynthesis.

Show MeSH
Related in: MedlinePlus