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E2f1 deletion attenuates infarct-induced ventricular remodeling without affecting O-GlcNAcylation

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Abstract

Several post-translational modifications figure prominently in ventricular remodeling. The beta-O-linkage of N-acetylglucosamine (O-GlcNAc) to proteins has emerged as an important signal in the cardiovascular system. Although there are limited insights about the regulation of the biosynthetic pathway that gives rise to the O-GlcNAc post-translational modification, much remains to be elucidated regarding the enzymes, such as O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), which regulate the presence/absence of O-GlcNAcylation. Recently, we showed that the transcription factor, E2F1, could negatively regulate OGT and OGA expression in vitro. The present study sought to determine whether E2f1 deletion would improve post-infarct ventricular function by de-repressing expression of OGT and OGA. Male and female mice were subjected to non-reperfused myocardial infarction (MI) and followed for 1 or 4 week. MI significantly increased E2F1 expression. Deletion of E2f1 alone was not sufficient to alter OGT or OGA expression in a naïve setting. Cardiac dysfunction was significantly attenuated at 1-week post-MI in E2f1-ablated mice. During chronic heart failure, E2f1 deletion also attenuated cardiac dysfunction. Despite the improvement in function, OGT and OGA expression was not normalized and protein O-GlcNAcyltion was not changed at 1-week post-MI. OGA expression was significantly upregulated at 4-week post-MI but overall protein O-GlcNAcylation was not changed. As an alternative explanation, we also performed guided transcriptional profiling of predicted targets of E2F1, which indicated potential differences in cardiac metabolism, angiogenesis, and apoptosis. E2f1 ablation increased heart size and preserved remote zone capillary density at 1-week post-MI. During chronic heart failure, cardiomyocytes in the remote zone of E2f1-deleted hearts were larger than wildtype. These data indicate that, overall, E2f1 exerts a deleterious effect on ventricular remodeling. Thus, E2f1 deletion improves ventricular remodeling with limited impact on enzymes regulating O-GlcNAcylation.

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Abbreviations

ANP:

Atrial natriuretic peptide

BCL2:

B-cell lymphoma 2

BNP:

Brain natriuretic peptide

CYTB:

Cytochrome b

E2F1:

E2F transcription factor 1

GFPT1:

Glutamine fructose-6-phosphate transaminase 1

GFPT2:

Glutamine fructose-6-phosphate transaminase 2

HBP:

Hexosamine biosynthetic pathway

MI:

Myocardial infarction

NDUFS1:

NADH: ubiquinone oxidoreductase core subunit S1

O-GlcNAc:

β-O-Linked N-acetylglucosamine

OGA:

O-GlcNAcase

OGT:

O-GlcNAc transferase

PDK1:

Pyruvate dehydrogenase kinase 1

PDK4:

Pyruvate dehydrogenase kinase 4

PFKFB1:

6-Phosphofructo-2-kinase/fructose-2,6-biphosphatase 1

PFKFB2:

6-Phosphofructo-2-kinase/fructose-2,6-biphosphatase 1

PGC1α:

Peroxisome proliferator-activated receptor gamma, co-activator 1 alpha

PGC1β:

Peroxisome proliferator-activated receptor gamma, co-activator 1 beta

VEGFA:

Vascular endothelial growth factor A

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Acknowledgements

The authors acknowledge the University of Louisville Diabetes and Obesity Center’s Imaging and Physiology Core.

Funding

Dr. Jones has been supported by Grants from the NIH (R01 HL131647, P30 GM127607, and P01 HL078825). Dr. Dassanayaka was supported by an American Heart Association Predoctoral Fellowship—Great Rivers Affiliate (14PRE19710015).

Author information

Authors and Affiliations

  1. Division of Cardiovascular Medicine, Department of Medicine, Diabetes and Obesity Center, Institute of Molecular Cardiology, University of Louisville, 580 South Preston Street-321F, Delia Baxter Building-321F, Louisville, KY, 40202, USA

    Sujith Dassanayaka, Kenneth R. Brittian, Andrea Jurkovic, Lauren A. Higgins, Timothy N. Audam, Bethany W. Long, Linda T. Harrison, Daniel W. Riggs, Mitali G. Chitre, Senthilkumar Muthusamy, Anna M. Gumpert & Steven P. Jones

  2. Division of Cardiovascular Medicine, Department of Medicine, Cardiovascular Innovation Institute, University of Louisville, Louisville, KY, USA

    Giuseppe Militello & Shizuka Uchida

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  1. Sujith Dassanayaka
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Contributions

SD performed experiments and analyzed data; wrote and revised manuscript. KRB performed experiments and analyzed data. AJ performed experiments and analyzed data. LAH performed experiments and analyzed data. TNA performed experiments and analyzed data. BWL performed experiments and analyzed data. LTH performed experiments and analyzed data. DWR analyzed data. GM performed experiments and analyzed data. MGC performed experiments and analyzed data. SU provided samples; revised manuscript. SM performed experiments and analyzed data. AMG performed experiments and analyzed data. SPJ designed experiments; wrote and revised manuscript.

Corresponding author

Correspondence to Steven P. Jones.

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Electronic supplementary material

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Supplementary material 1 (DOCX 63 kb)

395_2019_737_MOESM2_ESM.pdf

Supp Fig. 1: E2F1 deletion does not improve survival after MI. Survival of E2f1+/+ (n = 21) and E2f1−/− (n = 19) mice 1 week after MI (A). Survival of E2f1+/+ (n = 21) and E2f1−/− mice (n = 18) 4 week after MI (B). A log-rank test was used to determine significance between E2f1+/+ and E2f1−/− groups from Kaplan–Meier survival curves (PDF 39 kb)

395_2019_737_MOESM3_ESM.pdf

Supp Table 1: Attenuation of cardiac dysfunction is limited to male E2f1−/− mice. Male and female E2f1−/− (n = 25) and their E2f1+/+ (n = 26) littermates were subjected to echocardiography 1 week after MI. Indices of cardiac function in E2f1−/− and E2f1+/+ mice were separated by genotype and sex. E2f1−/− male mice preserved end-diastolic volume (EDV, p < 0.0001), end-systolic volume (ESV, p < 0.0001), stroke volume (SV, p < 0.01), ejection fraction (EF, p < 0.0001), left ventricular diastolic inner diameter (LVIDd, p < 0.001), left ventricular systolic inner diameter (LVIDs, p < 0.001), fractional shortening (FS, p < 0.01), and cardiac output (CO, p = 0226). There were no changes in interaction between gender and genotype in heart rate (HR), diastolic or systolic left ventricular anterior wall diameter (LVAWd, LVAWs), and diastolic or systolic left ventricular posterior wall diameter (LVPWd, LVPWs) between groups. A 2-way ANOVA with interaction was performed for genotype and gender followed by a Bonferroni post-test for each parameter of cardiac function (PDF 50 kb)

395_2019_737_MOESM4_ESM.pdf

Supp Fig. 2: Expression of Vegfa remains unaffected in acute and chronic heart failure. Vegfa mRNA expression at 1 and 4 week following MI (A and B, respectively). An unpaired Student’s t test was used to determine significance between E2f1+/+ and E2f1−/− groups (PDF 37 kb)

395_2019_737_MOESM5_ESM.pdf

Supp Table 2: Durable improvement in ventricular function is limited to male E2f1−/− mice. Male and female E2f1−/− (n = 15) and their E2f1+/+ (n = 17) littermates were subjected to echocardiography 4 week after MI. Indices of cardiac function in E2f1−/− and E2f1+/+ mice were separated by genotype and sex. E2f1−/− male mice preserved end-diastolic volume (EDV, p < 0.0001), end-systolic volume (ESV, p < 0.0001), stroke volume (SV, p = 0.0182), ejection fraction (EF, p = 0.0002), left ventricular diastolic inner diameter (LVIDd, p = 0.0006), left ventricular systolic inner diameter (LVIDs, p = 0.0008), fractional shortening (FS, p = 0.0076), left ventricular diastolic posterior wall diameter (LVPWd, p = 0.0467), and left ventricular systolic posterior wall diameter (LVPWs, p = 0.0189). There were no changes in interaction between gender and genotype in heart rate (HR), cardiac output (CO), and diastolic or systolic left ventricular anterior wall diameter (LVAWd, LVAWs). A 2-way ANOVA with interaction was performed for genotype and gender followed by a Bonferroni post-test for each parameter of cardiac function (PDF 50 kb)

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Dassanayaka, S., Brittian, K.R., Jurkovic, A. et al. E2f1 deletion attenuates infarct-induced ventricular remodeling without affecting O-GlcNAcylation. Basic Res Cardiol 114, 28 (2019). https://doi.org/10.1007/s00395-019-0737-y

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