Human induced pluripotent stem cell-derived cardiomyocytes to study inflammation-induced aberrant calcium transient
- Feinberg Cardiovascular and Renal Research Institute, Northwestern University, United States
- Department of Pharmacology, Northwestern University, United States
- Department of Medicine, Northwestern University, United States
- Department of Medicine, University of California, San Francisco, United States
Figures
Figure 1 with 2 supplements
Analyses of calcium transient in hiPSC-CMs treated with cytokines and/or MitoTempo.
Normalized downstroke time (A), decay time (B), and beating rate as assessed by beats per minute (C) in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) treated with bovine serum albumin (BSA), (control), TNF-α, IFN-γ, TNF-α plus mito-Tempo, and IFN-γ plus mito-Tempo. N = 5–14. Data were analyzed by ordinary one-way ANOVA and post hoc Tukey’s multiple-comparison test. Bars represent group mean, and error bars indicate standard error of the mean.
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Figure 1—source data 1
Analyses of calcium transient in hiPSC-CMs treated with cytokines and/or MitoTempo.
- https://cdn.elifesciences.org/articles/95867/elife-95867-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
A representative calcium transient curve in hiPSC-CMs and their mitochondrial potential.
(A) Representative image of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM). Scale bar represents 100 μm. (B) Representative curves of calcium transient and its derivative in hiPSC-CM. (C) Mitochondrial membrane potential (MMP) as assessed by tetramethylrhodamine ethyl ester (TMRE) in hiPSC-CM treated with various cytokines. Concentration of cytokines used is as follows: CCL3 1 nM, TNF-α 300 pg/ml, IL-6 1 ng/ml, IL-1β 200 pg/ml, IP-10 (or CXCL10) 500 pg/ml, IFN-γ 1 ng/ml. N = 4, Data were analyzed by ordinary one-way ANOVA and post hoc Tukey’s multiple-comparison test. Bars represent group mean , and error bars indicate standard error of the mean.
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Figure 1—figure supplement 1—source data 1
A epresentative image of hiPSC-CMs, a calcium transient curve, and a relative dF/dt curve.
- https://cdn.elifesciences.org/articles/95867/elife-95867-fig1-figsupp1-data1-v1.xlsx
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Figure 1—figure supplement 1—source data 2
Relative TMRE fluorescence in hiPSC-CMs with cytokine treatments.
- https://cdn.elifesciences.org/articles/95867/elife-95867-fig1-figsupp1-data2-v1.xlsx
Figure 1—video 1
Representative video of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) contracting spontaneously in a culture dish.
Scale bar represents 100 μm.
Figure 2
Oxygen consumption rate (OCR) in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) treated with cytokines and antioxidant agent N-acetylcysteine (NAC).
(A) OCR trace of hiPSC-CM treated with bovine serum albumin (BSA), TNF-α, IFN-γ, TNF-α + IFN-γ, TNF-α + NAC, IFN-γ + NAC, and TNF-α + IFN-γ + NAC. (B–D) Bar graph summary of data in (A) with OCR at baseline (B), after adding carbonyl cyanide m-chlorophenyl hydrazone (CCCP) (C), and after adding antimycin A and rotenone (D). N = 5–6. Data were analyzed by ordinary one-way ANOVA and post hoc Tukey’s multiple-comparison test. Bars represent group mean, and error bars indicate standard error of the mean.
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Figure 2—source data 1
OCR curve in hiPSCs treated with cytokines and NAC.
- https://cdn.elifesciences.org/articles/95867/elife-95867-fig2-data1-v1.xlsx
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Figure 2—source data 2
OCR at baseline, after CCCP treatment and after rotenone and antimycin A treatment.
- https://cdn.elifesciences.org/articles/95867/elife-95867-fig2-data2-v1.xlsx
Figure 3 with 1 supplement
Decay time in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) after treatment with TNF-α and various antiretroviral therapy (ART) drugs.
(A) Decay time after treatment with TNF-α, tenofovir, and TNF-α+tenofovir. (B) Decay time after treatment with TNF-α, darunavir, and TNF-α+darunavir. (C) Decay time after treatment with TNF-α, raltegravir, and TNF-α + raltegravir. (D) Decay time after treatment with TNF-α, emtricitabine, and TNF-α + emtricitabine. N = 5–21. Data were analyzed by ordinary one-way ANOVA and post hoc Tukey’s multiple-comparison test. Bars represent group mean, and error bars indicate standard error of the mean.
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Figure 3—source data 1
Normalized decay times in hiPSC-CMs treated with TNF-α and/or anti-HIV drugs.
- https://cdn.elifesciences.org/articles/95867/elife-95867-fig3-data1-v1.xlsx
Figure 3—figure supplement 1
Cell viability and ROS levels in hiPSC-CMs treated with anti-HIV drugs.
Cell viability (A) and cellular reactive oxygen species (ROS) levels (B) in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) exposed to concentrations ranging from between 3 nM and 10 μM of tenofovir (a nucleotide-analog reverse transcriptase inhibitor), darunavir (a protease inhibitor), raltegravir, and elvitegravir (integrase inhibitors). N=1-6.
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Figure 3—figure supplement 1—source data 1
Cell viability and ROS levels in hiPSC-CMs treated with anti-HIV drugs.
- https://cdn.elifesciences.org/articles/95867/elife-95867-fig3-figsupp1-data1-v1.xlsx
Figure 4 with 1 supplement
Decay time in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) after treatment with TNF-α and riociguat, sildenafil citrate, PF-04447943, and dapagliflozin.
(A) Decay time after treatment with TNF-α, sGS agonist riociguat, and TNF-α + riociguat (N = 8–21). (B) Decay time after treatment with TNF-α, sildenafil citrate, and TNF-α + sildenafil citrate (N = 5–21). (C) Decay time after treatment with TNF-α, PF-04447943, and TNF-α+PF-04447943 (N = 7–21). (D) Representative curve of calcium transient in hiPSC-CM treated with TNF-α, dapagliflozin, and TNF-α + dapagliflozin. (E) Curve of derivative calculated with data in (D). (F) Decay time after treatment with TNF-α, dapagliflozin, and TNF-α+dapagliflozin (N = 5–21). (G) Downstroke time after treatment with TNF-α, dapagliflozin, and TNF-α + dapagliflozin(N = 5–21). (H) Representative curve of calcium transient in hiPSC-CM treated with IFN-γ, dapagliflozin, and IFN-γ + dapagliflozin. (I) Curve of derivative calculated with data in (H). (J) Decay time after treatment with INF-γ, dapagliflozin, and IFN-γ + dapagliflozin (N = 5–21). (K) Downstroke time after treatment with INF-γ, dapagliflozin, and IFN-γ + dapagliflozin (N = 5–21). Data were analyzed by ordinary one-way ANOVA and post hoc Tukey’s multiple-comparison test. Bars represent group mean, and error bars indicate standard error of the mean.
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Figure 4—source data 1
Normalizaed decay times in hiPSC-CMs treated with TNFα and Riociguat, Sildenafil citrate and PF-04447943.
- https://cdn.elifesciences.org/articles/95867/elife-95867-fig4-data1-v1.xlsx
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Figure 4—source data 2
Analyses of calcium transient in hiPSC-CMs treated with TNFα, INFγ and 10 μM Dapagliflozin.
- https://cdn.elifesciences.org/articles/95867/elife-95867-fig4-data2-v1.xlsx
Figure 4—figure supplement 1
Analyses of calcium transient in hiPSC-CMs treated with TNFa and/or 1 μM Dapagliflozin.
Decay time (A) and downstroke time (B) in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) after treatment with TNF-α and 1 μM of dapagliflozin.
N = 5–21, Data were analyzed by ordinary one-way ANOVA and post hoc Tukey’s multiple-comparison test. Bars represent group mean, and error bars indicate standard error of the mean.
Figure 5 with 2 supplements
Ca2+-decay time of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) and angiogenic function of ECs after treatment with serum from group 1 HIV+ patients with diastolic dysfunction (DD).
(A–C) Pooled normalized decay time (A), downstroke time (B), and beating rate (C) in hiPSC treated with 10% serum for 48 hr from group 1 HIV+ patients with DD (3 values from cells treated with each serum). (D–G) Angiogenic parameters, including total length of branching (D) (N = 3), number of junctions (E) (N = 3), total length (F) (N = 3), and number of meshes (G) (N = 3) in human umbilical vein endothelial cells (HUVEC) after treatment with 2% serum from group 1 patients for 20 hr. Data were analyzed by unpaired Student’s t-test. Bars represent group mean, and error bars indicate standard error of the mean.
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Figure 5—source data 1
Analyses of calcium transient in hiPSCs treated with serum from HIV patients.
- https://cdn.elifesciences.org/articles/95867/elife-95867-fig5-data1-v1.xlsx
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Figure 5—source data 2
Quantitative analyses of tube formation assay in HUVECs treated with serum from HIV patients.
- https://cdn.elifesciences.org/articles/95867/elife-95867-fig5-data2-v1.xlsx
Figure 5—figure supplement 1
Representative images of formed tube structure with human umbilical vein endothelial cells (HUVEC) treated with 2% serum from HIV+ patients with high or low myocardial perfusion reserve (MPR) for 20 hr.
Scale bars represent 500 μm.
Figure 5—figure supplement 2
Representative magnified images of formed tube structure with human umbilical vein endothelial cells (HUVEC) treated with 2% serum from HIV+ patients with high or low myocardial perfusion reserve (MPR) for 20 hr.
Scale bars indicate 250 μm.
Figure 6
Diastolic function of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) and angiogenic function of ECs after treatment with serum from HIV+ patients with diastolic dysfunction (DD) (group 2).
(A, B) Individual normalized decay time 3 hr (N = 3–9) (A) or 24 hr (N = 3–13) (B) after treatment of hiPSC-CM with 2% serum from patients. (C, D) Pooled decay time data from patients in (A) and (B) with 3 hr (C) and 24 hr (D) after treatment with serum (N = 21–22 for (C) and N = 31-38 for (D)). (E) Representative image of formed tube structure with cultured human umbilical vein endothelial cells (HUVEC) treated with serum from HIV+ patients with DD. Scale bars indicate 250 μm. (F–I) Assessment of tube formation of HUVECs 20 hr after treatment with serum of patients, including number of meshes (F) (N = 8), number of junctions (G) (N = 8), number of segments (H) (N = 8), and total length of branching (I) (N = 8). Data were analyzed by unpaired Student’s t-test for (C, D, F, G, H, I). Bars represent group mean, and error bars indicate standard error of the mean.
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Figure 6—source data 1
Normalized decay time in iPS-CMs treated with HIV patient serum with or without diastolic dysfunction.
- https://cdn.elifesciences.org/articles/95867/elife-95867-fig6-data1-v1.xlsx
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Figure 6—source data 2
Quantitative analyses of tube formation assay in HUVECs treated with serum from HIV patients.
- https://cdn.elifesciences.org/articles/95867/elife-95867-fig6-data2-v1.xlsx
Tables
Table 1
Clinical characteristics of study population for group 1.
| Participant | Age | High or low myocardial perfusion reserve group | Sex | CD4 count | Diabetes mellitus? | Active cigarette smoking? | Hypertension? | Coronary artery disease diagnosis? | Left ventricular ejection fraction | Myocardial perfusion reserve |
|---|---|---|---|---|---|---|---|---|---|---|
| 1 | 50 | High | Male | 487 | No | Yes | Yes | No | 64 | 2.74 |
| 2 | 62 | Low | Male | 703 | No | No | Yes | Yes | 59 | 1.03 |
| 3 | 64 | High | Male | 321 | No | No | No | Yes | 65 | 3.43 |
| 4 | 55 | High | Male | 749 | No | No | Yes | Yes | 59 | 1.90 |
| 5 | 71 | Low | Male | 1743 | No | No | Yes | Yes | 55 | 1.03 |
| 6 | 59 | Low | Male | 678 | No | No | Yes | No | 52 | 1.13 |
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Human induced pluripotent stem cell-derived cardiomyocytes to study inflammation-induced aberrant calcium transient
eLife 13:RP95867.
https://doi.org/10.7554/eLife.95867.3
