FOXO3a regulates BNIP3 and modulates mitochondrial calcium, dynamics, and function in cardiac stress.
The forkhead box O3a (FOXO3a) transcription factor has been shown to regulate glucose metabolism, muscle atrophy, and cell death in postmitotic cells. Its role in regulation of mitochondrial and myocardial function is not well studied. Based on previous work, we hypothesized that FOXO3a, through BCL2/adenovirus E1B 19-kDa protein-interacting protein 3 (BNIP3), modulates mitochondrial morphology and function in heart failure (HF). We modulated the FOXO3a-BNIP3 pathway in normal and phenylephrine (PE)-stressed adult cardiomyocytes (ACM) in vitro and developed a cardiotropic adeno-associated virus serotype 9 encoding dominant-negative FOXO3a (AAV9.dn-FX3a) for gene delivery in a rat model of HF with preserved ejection fraction (HFpEF). We found that FOXO3a upregulates BNIP3 expression in normal and PE-stressed ACM, with subsequent increases in mitochondrial Ca2+, leading to decreased mitochondrial membrane potential, mitochondrial fragmentation, and apoptosis. Whereas dn-FX3a attenuated the increase in BNIP3 expression and its consequences in PE-stressed ACM, AAV9.dn-FX3a delivery in an experimental model of HFpEF decreased BNIP3 expression, reversed adverse left ventricular remodeling, and improved left ventricular systolic and, particularly, diastolic function, with improvements in mitochondrial structure and function. Moreover, AAV9.dn-FX3a restored phospholamban phosphorylation at S16 and enhanced dynamin-related protein 1 phosphorylation at S637. Furthermore, FOXO3a upregulates maladaptive genes involved in mitochondrial apoptosis, autophagy, and cardiac atrophy. We conclude that FOXO3a activation in cardiac stress is maladaptive, in that it modulates Ca2+cycling, Ca2+homeostasis, and mitochondrial dynamics and function. Our results suggest an important role of FOXO3a in HF, making it an attractive potential therapeutic target.
Division of Cardiovascular Diseases, Mayo Clinic, Rochester, Minnesota.
Antoine H
0000-0002-9603-7849
AH
Chaanine
Division of Cardiovascular Diseases, Mount Sinai School of Medicine, New York, New York.
Erik
E
Kohlbrenner
Microscopy and Cell Analysis Core, Department of Biochemistry and Cell Biology, Mayo Clinic, Rochester, Minnesota; and.
Scott I
SI
Gamb
Division of Cardiovascular Diseases, Mayo Clinic, Rochester, Minnesota.
Adam J
AJ
Guenzel
Division of Endocrinology, Mayo Clinic, Rochester, Minnesota.
Katherine
K
Klaus
Division of Cardiovascular Diseases, Mayo Clinic, Rochester, Minnesota.
Ahmed U
AU
Fayyaz
Division of Endocrinology, Mayo Clinic, Rochester, Minnesota.
K Sreekumaran
KS
Nair
Division of Cardiovascular Diseases, Mount Sinai School of Medicine, New York, New York.
Roger J
RJ
Hajjar
Division of Cardiovascular Diseases, Mayo Clinic, Rochester, Minnesota; redfield.margaret@mayo.edu.
Margaret M
MM
Redfield
D000818:Animals; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002470:Cell Survival; D002950:Citrate (si)-Synthase / Q000378:metabolism; D004195:Disease Models, Animal; D034281:Dynamins / Q000378:metabolism; D004452:Echocardiography; D003576:Electron Transport Complex IV / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D000071316:Forkhead Box Protein O3 / Q000235:genetics* / Q000378:metabolism; D006333:Heart Failure / Q000378:metabolism* / Q000503:physiopathology; D066298:In Vitro Techniques; D008297:Male; D053078:Membrane Potential, Mitochondrial; D008565:Membrane Proteins / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D024101:Mitochondrial Proteins / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D010656:Phenylephrine / Q000494:pharmacology; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D013312:Stress, Physiological; D013318:Stroke Volume; D013566:Sympathomimetics / Q000494:pharmacology; D016277:Ventricular Function, Left / Q000235:genetics; D020257:Ventricular Remodeling; D000099211:Phospholamban
D002118:Calcium; D002135:Calcium-Binding Proteins; D002950:Citrate (si)-Synthase; D034281:Dynamins; D003576:Electron Transport Complex IV; D000071316:Forkhead Box Protein O3; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D010656:Phenylephrine; D013566:Sympathomimetics; D000099211:Phospholamban; C505650:BNIP3 protein, rat; C499515:FOXO3 protein, rat; C519851:Dnm1l protein, rat
BNIP3; FOXO3a; apoptosis; calcium regulation; heart failure
J Biol Chem. 2008 Mar 28;283(13):8211-7
18195003
Physiol Genomics. 2002;9(1):49-56
11948290
J Mol Cell Cardiol. 2008 Aug;45(2):270-80
18597775
Cell Death Dis. 2013 May 09;4:e625
23661003
J Biol Chem. 2014 Apr 11;289(15):10812-22
24567336
Am J Physiol Gastrointest Liver Physiol. 2012 Apr;302(7):G723-31
22241863
Oncogene. 2005 Nov 14;24(50):7410-25
16288288
Proc Natl Acad Sci U S A. 2008 Oct 14;105(41):15803-8
18838687
J Cell Biol. 2003 Aug 18;162(4):613-22
12913110
J Biol Chem. 2004 Aug 13;279(33):34741-9
15184386
Cell Metab. 2007 Dec;6(6):458-71
18054315
Circ Heart Fail. 2013 May;6(3):572-83
23508759
J Biol Chem. 2005 May 27;280(21):20814-23
15781459
Cell Death Dis. 2012 Feb 02;3:265
22297293
Eur J Heart Fail. 2010 Dec;12 (12 ):1290-6
21098579
Science. 2002 Apr 19;296(5567):530-4
11964479
J Biol Chem. 2007 Jul 27;282(30):21583-7
17553808
Proc Natl Acad Sci U S A. 2015 Sep 8;112(36):11389-94
26217001
Mol Ther. 2012 Jan;20(1):73-83
21915102
Mol Biol Cell. 2009 Feb;20(3):1058-67
19037102
Biochim Biophys Acta. 2011 Jul;1812(7):743-51
21447388
Proc Natl Acad Sci U S A. 2004 Mar 2;101(9):2975-80
14978268
Cardiovasc J Afr. 2010 Jan-Feb;21(1):37-41
20224844
Int J Biochem Cell Biol. 2009 Oct;41(10):1957-71
19703657
Prostate. 2008 Feb 15;68(3):336-43
18163427
Annu Rev Physiol. 2010;72:81-94
20148668
Nat Rev Cardiol. 2014 Sep;11(9):507-15
24958077
J Clin Invest. 2009 Sep;119(9):2758-71
19652361
Biomed Res Int. 2014;2014:925350
24864265
Cell Death Differ. 2011 Dec;18(12):1914-23
21637291
Am J Physiol Heart Circ Physiol. 2014 Mar;306(6):H844-55
24441545
J Mol Cell Cardiol. 2009 Jun;46(6):781-8
19285982
Physiology (Bethesda). 2008 Apr;23:84-94
18400691
Shock. 2011 Aug;36(2):196-202
21558981
Eur J Clin Invest. 1995 Aug;25(8):548-52
7589009
Mol Cell Biol. 2009 May;29(10 ):2570-81
19273585
Oncogene. 2008 Oct 27;27(50):6407-18
18955969
J Cell Sci. 2009 Apr 1;122(Pt 7):1005-13
19295127
Nat Protoc. 2008;3(9):1422-34
18772869
Biochem J. 1976 Mar 15;154(3):689-700
8036
Biochim Biophys Acta. 2009 Nov;1787(11):1395-401
19576166
Mol Biol Cell. 2002 Sep;13(9):3355-68
12221139
J Exp Med. 2006 Jul 10;203(7):1657-63
16801400