FoxO1 regulates myocardial glucose oxidation rates via transcriptional control of pyruvate dehydrogenase kinase 4 expression

Keshav Gopal, Bruno Saleme, Rami Al Batran, Hanin Aburasayn, Amina Eshreif, Kim L. Ho, Wayne K. Ma, Malak Almutairi, Farah Eaton, Manoj Gandhi, Edwards Park, Gopinath Sutendra, John R. Ussher

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Abstract

Pyruvate dehydrogenase (PDH) is the ratelimiting enzyme for glucose oxidation and a critical regulator of metabolic flexibility during the fasting to feeding transition. PDH is regulated via both PDH kinases (PDHK) and PDH phosphatases, which phosphorylate/inactivate and dephosphorylate/activate PDH, respectively. Our goal was to determine whether the transcription factor forkhead box O1 (FoxO1) regulates PDH activity and glucose oxidation in the heart via increasing the expression of Pdk4, the gene encoding PDHK4. To address this question, we differentiated H9c2 myoblasts into cardiac myocytes and modulated FoxO1 activity, after which Pdk4/PDHK4 expression and PDH phosphorylation/activity were assessed. We assessed binding of FoxO1 to the Pdk4 promoter in cardiac myocytes in conjunction with measuring the role of FoxO1 on glucose oxidation in the isolated working heart. Both pharmacological (1 µM AS1842856) and genetic (siRNA mediated) inhibition of FoxO1 decreased Pdk4/PDHK4 expression and subsequent PDH phosphorylation in H9c2 cardiac myocytes, whereas 10 µM dexamethasone-induced Pdk4/PDHK4 expression was abolished via pretreatment with 1 µM AS1842856. Furthermore, transfection of H9c2 cardiac myocytes with a vector expressing FoxO1 increased luciferase activity driven by a Pdk4 promoter construct containing the FoxO1 DNA-binding element region, but not in a Pdk4 promoter construct lacking this region. Finally, AS1842856 treatment in fasted mice enhanced glucose oxidation rates during aerobic isolated working heart perfusions. Taken together, FoxO1 directly regulates Pdk4 transcription in the heart, thereby controlling PDH activity and subsequent glucose oxidation rates. New & Noteworthy Although studies have shown an association between FoxO1 activity and pyruvate dehydrogenase kinase 4 expression, our study demonstrated that pyruvate dehydrogenase kinase 4 is a direct transcriptional target of FoxO1 (but not FoxO3/FoxO4) in the heart. Furthermore, we report here, for the first time, that FoxO1 inhibition increases glucose oxidation in the isolated working mouse heart.

Original languageEnglish (US)
Pages (from-to)H479-H490
JournalAmerican Journal of Physiology - Heart and Circulatory Physiology
Volume313
Issue number3
DOIs
StatePublished - Jan 1 2017

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Pyruvic Acid
Oxidoreductases
Glucose
Cardiac Myocytes
Pyruvate Dehydrogenase (Lipoamide)-Phosphatase
Phosphorylation
Glucose 1-Dehydrogenase
Forkhead Transcription Factors
Myoblasts
pyruvate dehydrogenase kinase 4
Luciferases
Dexamethasone
Small Interfering RNA
Transfection
Fasting
Perfusion
Pharmacology
Gene Expression
DNA
Enzymes

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

FoxO1 regulates myocardial glucose oxidation rates via transcriptional control of pyruvate dehydrogenase kinase 4 expression. / Gopal, Keshav; Saleme, Bruno; Batran, Rami Al; Aburasayn, Hanin; Eshreif, Amina; Ho, Kim L.; Ma, Wayne K.; Almutairi, Malak; Eaton, Farah; Gandhi, Manoj; Park, Edwards; Sutendra, Gopinath; Ussher, John R.

In: American Journal of Physiology - Heart and Circulatory Physiology, Vol. 313, No. 3, 01.01.2017, p. H479-H490.

Research output: Contribution to journalArticle

Gopal, K, Saleme, B, Batran, RA, Aburasayn, H, Eshreif, A, Ho, KL, Ma, WK, Almutairi, M, Eaton, F, Gandhi, M, Park, E, Sutendra, G & Ussher, JR 2017, 'FoxO1 regulates myocardial glucose oxidation rates via transcriptional control of pyruvate dehydrogenase kinase 4 expression', American Journal of Physiology - Heart and Circulatory Physiology, vol. 313, no. 3, pp. H479-H490. https://doi.org/10.1152/ajpheart.00191.2017
Gopal, Keshav ; Saleme, Bruno ; Batran, Rami Al ; Aburasayn, Hanin ; Eshreif, Amina ; Ho, Kim L. ; Ma, Wayne K. ; Almutairi, Malak ; Eaton, Farah ; Gandhi, Manoj ; Park, Edwards ; Sutendra, Gopinath ; Ussher, John R. / FoxO1 regulates myocardial glucose oxidation rates via transcriptional control of pyruvate dehydrogenase kinase 4 expression. In: American Journal of Physiology - Heart and Circulatory Physiology. 2017 ; Vol. 313, No. 3. pp. H479-H490.
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abstract = "Pyruvate dehydrogenase (PDH) is the ratelimiting enzyme for glucose oxidation and a critical regulator of metabolic flexibility during the fasting to feeding transition. PDH is regulated via both PDH kinases (PDHK) and PDH phosphatases, which phosphorylate/inactivate and dephosphorylate/activate PDH, respectively. Our goal was to determine whether the transcription factor forkhead box O1 (FoxO1) regulates PDH activity and glucose oxidation in the heart via increasing the expression of Pdk4, the gene encoding PDHK4. To address this question, we differentiated H9c2 myoblasts into cardiac myocytes and modulated FoxO1 activity, after which Pdk4/PDHK4 expression and PDH phosphorylation/activity were assessed. We assessed binding of FoxO1 to the Pdk4 promoter in cardiac myocytes in conjunction with measuring the role of FoxO1 on glucose oxidation in the isolated working heart. Both pharmacological (1 µM AS1842856) and genetic (siRNA mediated) inhibition of FoxO1 decreased Pdk4/PDHK4 expression and subsequent PDH phosphorylation in H9c2 cardiac myocytes, whereas 10 µM dexamethasone-induced Pdk4/PDHK4 expression was abolished via pretreatment with 1 µM AS1842856. Furthermore, transfection of H9c2 cardiac myocytes with a vector expressing FoxO1 increased luciferase activity driven by a Pdk4 promoter construct containing the FoxO1 DNA-binding element region, but not in a Pdk4 promoter construct lacking this region. Finally, AS1842856 treatment in fasted mice enhanced glucose oxidation rates during aerobic isolated working heart perfusions. Taken together, FoxO1 directly regulates Pdk4 transcription in the heart, thereby controlling PDH activity and subsequent glucose oxidation rates. New & Noteworthy Although studies have shown an association between FoxO1 activity and pyruvate dehydrogenase kinase 4 expression, our study demonstrated that pyruvate dehydrogenase kinase 4 is a direct transcriptional target of FoxO1 (but not FoxO3/FoxO4) in the heart. Furthermore, we report here, for the first time, that FoxO1 inhibition increases glucose oxidation in the isolated working mouse heart.",
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AU - Saleme, Bruno

AU - Batran, Rami Al

AU - Aburasayn, Hanin

AU - Eshreif, Amina

AU - Ho, Kim L.

AU - Ma, Wayne K.

AU - Almutairi, Malak

AU - Eaton, Farah

AU - Gandhi, Manoj

AU - Park, Edwards

AU - Sutendra, Gopinath

AU - Ussher, John R.

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N2 - Pyruvate dehydrogenase (PDH) is the ratelimiting enzyme for glucose oxidation and a critical regulator of metabolic flexibility during the fasting to feeding transition. PDH is regulated via both PDH kinases (PDHK) and PDH phosphatases, which phosphorylate/inactivate and dephosphorylate/activate PDH, respectively. Our goal was to determine whether the transcription factor forkhead box O1 (FoxO1) regulates PDH activity and glucose oxidation in the heart via increasing the expression of Pdk4, the gene encoding PDHK4. To address this question, we differentiated H9c2 myoblasts into cardiac myocytes and modulated FoxO1 activity, after which Pdk4/PDHK4 expression and PDH phosphorylation/activity were assessed. We assessed binding of FoxO1 to the Pdk4 promoter in cardiac myocytes in conjunction with measuring the role of FoxO1 on glucose oxidation in the isolated working heart. Both pharmacological (1 µM AS1842856) and genetic (siRNA mediated) inhibition of FoxO1 decreased Pdk4/PDHK4 expression and subsequent PDH phosphorylation in H9c2 cardiac myocytes, whereas 10 µM dexamethasone-induced Pdk4/PDHK4 expression was abolished via pretreatment with 1 µM AS1842856. Furthermore, transfection of H9c2 cardiac myocytes with a vector expressing FoxO1 increased luciferase activity driven by a Pdk4 promoter construct containing the FoxO1 DNA-binding element region, but not in a Pdk4 promoter construct lacking this region. Finally, AS1842856 treatment in fasted mice enhanced glucose oxidation rates during aerobic isolated working heart perfusions. Taken together, FoxO1 directly regulates Pdk4 transcription in the heart, thereby controlling PDH activity and subsequent glucose oxidation rates. New & Noteworthy Although studies have shown an association between FoxO1 activity and pyruvate dehydrogenase kinase 4 expression, our study demonstrated that pyruvate dehydrogenase kinase 4 is a direct transcriptional target of FoxO1 (but not FoxO3/FoxO4) in the heart. Furthermore, we report here, for the first time, that FoxO1 inhibition increases glucose oxidation in the isolated working mouse heart.

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