Glucose inhibits cardiac muscle maturation through nucleotide biosynthesis
- University of California, Los Angeles, United States
- Kyoto University, Japan
- National Institute for Materials Science, Japan
Figures
Figure 1
High yield hESC-CMs derived under chemically defined conditions recapitulate developmental time course.
(A) Representative flow cytometry analysis of a cardiac marker, MF20 (myosin) in hESC-CMs (H9) at day 14. (B) Bar chart of the percentage of MF20+ cardiomyocytes in hESC-CMs generated from H9 (n = 8)…
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Figure 1—source data 1
The contents of hESC-CM differentiation media.
- https://doi.org/10.7554/eLife.29330.004
Figure 2 with 1 supplement
Glucose reduction promotes maturation of hESC-CMs.
(A) Experimental regimen. hESC-CMs are differentiated in the medium containing 25 mM glucose until day 14, when ~ 90% of the cells are already MF20+. Cells are analyzed at day 28 unless otherwise …
Figure 2—figure supplement 1
Glucose reduction promotes maturation of hESC-CMs.
(A) Expression of cardiac markers extracted from RNA-seq data. (n = 3, mean ±S D, by t-test). (B) Sarcomere length analysis. Representative traces of α-actinin and cellular architecture (left) and …
Figure 3 with 1 supplement
Glucose deprivation promotes the functional maturation of hESC-CM.
(A) Representative images of mitochondrial membrane potential assay using JC-1 dye in hESC-CMs cultured in the presence (left) and absence (right) of glucose. Note the elongated mitochondria in the …
Figure 3—figure supplement 1
Electrophysiological analyses of the maturity of hESC-CMs by multi-electrode array (MEA).
(A) Maximum dv/dt of field potential obtained from three independent measurements of channels properly recorded by 120 electrodes with representative trace of recorded field potential; recorded …
Figure 4 with 4 supplements
The pentose phosphate pathway inhibits cardiac maturation.
(A) Heatmap presentation of the metabolomics analysis of hESC-CMs cultured in the presence or absence of glucose. Note the decrease in the metabolites in purine metabolism, pyrimidine metabolism and …
Figure 4—figure supplement 1
Metabolomics analyses by mass spectrometry.
The top diagram shows the comparison of metabolomics analysis by mass spectrometry of hESC-CMs cultured in the presence or absence of glucose. Each hexagon indicates a metabolite that is decreased …
Figure 4—figure supplement 2
RNAi knockdown of glucose metabolic enzymes.
Relative TNNT2 expression after RNA interference (RNAi) knockdown. RNAi targeting scramble, HK1, RRM2, RRM2B, G6PD, and PFK were transfected by lipofection for 48 hr followed by 7 days’ incubation …
Figure 4—figure supplement 3
The pentose phosphate pathway inhibits cardiac maturation.
Summary of the impact of glucose metabolism inhibitors on cardiac maturity. The inhibitors tested are shown in red boxes, and the effective inhibitors are highlighted in yellow. There is clear eviden…
Figure 4—figure supplement 4
Summary of the impact of glucose metabolism inhibitors on cardiac maturity.
(A) Relative mRNA expression of TNNT2 and NKX2-5 in conditions of 0–25 mM glucose and 0–25 mM 2-DG. No data were available for the samples with 2-DG concentration higher than its glucose level …
Figure 5 with 2 supplements
Nucleotide metabolism regulates cardiomyocyte maturation.
(A) Glucose-deprived hESC-CMs are cultured in the absence (a, b) or presence (c, d) of 25 mM uridine, and stained for pH3 (a mitosis marker). The addition of uridine restored proliferative activity …
Figure 5—figure supplement 1
Nucleotide inhibits hESC-CM maturation.
(A) The relative expression of TNNT2 mRNA measured by qPCR in hESC-CMs in 16 different conditions of 0–25 mM of glucose and 0–25 mM uridine. Uridine recapitulates the effect of glucose by …
Figure 5—figure supplement 2
Nucleotide deprivation, not cell cycle block, is the primary inducer of cardiomyocyte maturation.
(A) The impact of CDK4/6 inhibitor on the relative mRNA expression of TNNT2 and NKX2-5 measured by qPCR. Cell cycle block by CDK4/6 inhibitor did not significantly induce the expression of cardiac …
Figure 6
Developmental time course of cardiac glucose uptake measured by 18F-FDG accumulation.
The radioactivity of the entire heart was measured by γ-counter after tail vein i.v. (fetus) or i.p. (neonates) injections. Values were normalized to heart weight and total body signal (heart …
Figure 7
Hyperglycemia promotes the proliferation and inhibits the maturation of fetal cardiomyocytes in utero.
(A) Diagram illustrating in vivo analysis of the impact of maternal hyperglycemia on fetal heart development using a diabetic mouse model (Akita). (B,C) Cell cycle analyses of fetal and neonatal …
Author response image 1
Author response image 2
Videos
Video 1
Beating hESC-CMs differentiation in glucose 25 mM medium.
This video shows beating of hESC-CMs differentiated in the medium containing 25 mM glucose from day 14 for 7 days.
Video 2
Beating hESC-CMs differentiation in glucose 0 mM medium.
This video shows beating of hESC-CMs differentiated in the medium containing 0 mM glucose from day 14 for 7 days.
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.29330.021
