Modeling susceptibility to drug-induced long QT with a panel of subject-specific induced pluripotent stem cells
- Icahn School of Medicine at Mount Sinai, United States
- Sorbonne Universités, UPMC Univ Paris 06, AP-HP, INSERM, CIC-1421, Institute of Cardiometabolism and Nutrition, France
- Cellectis Stem Cells, France
Figures
Figure 1 with 1 supplement
QTcf changes following Sotalol administration in healthy volunteers.
(A) Flow chart of the clinical study. (B) Distribution of QTcf duration before (blue) and 3 hr after sotalol intake (red). (C) Distribution of delta change in QTcf showing the wide inter-individual …
Figure 1—figure supplement 1
Overview of the clinical and experimental study and role of each partner.
This translational study started with a prospective clinical study performed in a clinical investigation center near Paris, France. The objective was to identify healthy subjects with extreme …
Figure 2 with 3 supplements
Expression of sarcomeric proteins and ion channels in human iPSC-CMs.
(A) Confocal microscopy imaging of Troponin T (top), alpha-actinin and connexin 43 (bottom) in single generated iPSC-CMs (from line P11015). Nuclei are stained with DAPI (Blue). (B) Gene expression …
-
Figure 2—source data 1
iPSCs characterization.
Representative immunostaining for a panel of pluripotent stem cell markers, including Nanog, OCT4, SSEA4 and TRA1-60. Results are reported for each cell line. Typical staining in hESC is reported as a positive control. Scale bar 50 μM.
- https://doi.org/10.7554/eLife.19406.008
-
Figure 2—source data 2
Expression of main human cardiac ion channels.
Quantitative PCR results for the expression of 10 major cardiac ion channels including SCN5A, CACNA1, KCNQ1, KCNE1, KCNJ2, KCNJ11, KChIP2, HCN1, HCN2, HCN4. KCND3 was not detected. Adult LV tissue is used as a positive control and the level of expression in human embryonic stem cells-derived cardiomyocytes as a comparator.
- https://doi.org/10.7554/eLife.19406.009
Figure 2—figure supplement 1
Expression of exogenous and endogenous pluripotency genes.
Representative gel showing silencing of the four exogenous reprogramming transgenes (OCT3/4; KLF4; SOX2 ; c-MYC) and quantitative PCR results for endogenous pluripotent stem cell genes (OCT3/4; …
Figure 2—figure supplement 2
MEA arrays.
(A) Picture of a 6-well MEA array as used in the study. Each well contains nine micro-electrodes and a limited volume capacity allowing easier contact between cells and micro-electrodes after …
Figure 2—figure supplement 3
Quality of FPD adjustment.
(A) Plot of adjusted FPD using the Bazett’s formula against the individual inter-beat interval. This plot shows the miss-correction achieved using the Bazett’s formula with a significant dependence …
Figure 3
Differences in iPSC-CMs responses to Sotalol stimulation according to clinical sensitivity to Sotalol.
(A) Adjusted FPD (aFPD) measured in iPSC-CMs derived from subjects with low-sensitivity (green) vs. high-sensitivity (red) in response to increasing concentrations of sotalol. aFPD are normalized to …
Figure 4
Patch-clamp analysis of action potential (AP) in representative iPSC-CMs.
(A) Representative AP tracings of the iPSC-CMs generated from the low-S (P11009) and high-S (P11029) lines in control and sotalol-treated conditions. (B) bar chart summarizing the APD90 in control …
Figure 5 with 1 supplement
Identification of dysregulated genes as direct neighbors of QT-associated network in high-S iPSC-CMs.
(A) Known LQTS genes were used as seed nodes (green squares) in the human interactome and five differentially expressed direct neighbors were identified (circles) (path length 1). Up-regulated genes …
Figure 5—figure supplement 1
Prediction of sex hormones-related transcription factors.
Predicted regulatory transcription factors of the identified DEGs (DLG2, KCNE4, CAMKV, PTRF, HTR2C) based on two transcription factor target databases (Chea background and Transfac) and ranked by …
Author response image 1
Videos
Video 1
Spontaneous beating activity of monolayer iPSC-CMs seeded onto one well of a 6-well MEA chamber.
https://doi.org/10.7554/eLife.19406.014Tables
Table 1
Related to Figure 1. Demographic, clinical and electrocardiographic baseline characteristics of subjects with low- vs. high- sensitivity to Sotalol.
Low-sensitivity, n = 10 | High- sensitivity, n = 10 | p-value | |
|---|---|---|---|
Age | 29.9 ± 5.9 | 28.3 ± 5.9 | 0.53 |
Gender, Male (%) | 8 (80.0%) | 1 (10%) | 0.006 |
Body Mass Index (kg/m²) | 23.6 ± 0.9 | 22.9 ± 2.7 | 0.40 |
SBP (mmHg) | 116.0 ± 8.6 | 113.7 ± 8.3 | 0.50 |
DBP (mmHg) | 72.2 ± 4.8 | 68.0 ± 4.2 | 0.07 |
Resting heart Rate (bpm) | 59.6 ± 5.9 | 63.2 ± 8.4 | 0.37 |
PR interval (ms) | 166.7 ± 24.7 | 143.5 ± 18.1 | 0.09 |
QRS (ms) | 88.3 ± 5.9 | 85.6 ± 7.7 | 0.08 |
QTcf (ms) | 384.7 ± 26.4 | 402.6 ± 20.8 | 0.13 |
Table 2
Related to Figure 2. Fibroblasts and iPS quality control parameters.
ID | Mycoplasma* | HIV*, HBV*, HCV*, HTLV1 and 2* | Phosphatase alkaline staining | Karyotyping |
|---|---|---|---|---|
P11007~5924~iPSpolyRoksmA | Negative | Negative | Positive | 46,XY |
P11008~5444~iPSpolyRoksmB | Negative | Negative | Positive | 46, XX |
P11009~6426~iPSpolyRoksmC | Negative | Negative | Positive | 46, XY |
P11013~5744~iPSpolyRoksmD | Negative | Negative | Positive | 46, XX |
P11014-5864-iPSpolyRoksmC | Negative | Negative | Positive | 46, XX |
P11015~6345~iPSpolyRoksmE | Negative | Negative | Positive | 46, XX |
P11018~5644~iPSpolyRoksmB | Negative | Negative | Positive | 46, XX |
P11019-6444-iPSpolyRoksmB | Negative | Negative | Positive | 46, XY |
P11020 ~ 7125 ~ iPSpolyRoksmD | Negative | Negative | Positive | 46, XY |
P11021 ~ 6544 ~ iPSpolyRoksmC | Negative | Negative | Positive | 46, XX |
P11023~5525~iPSpolyRoksmA | Negative | Negative | Positive | 46, XX |
P11024~5844~iPSpolyRoksmA | Negative | Negative | Positive | 46, XX |
P11026~6504~iPSpolyRoksmD | Negative | Negative | Positive | 46, XY |
P11028~6904~iPSpolyRoksmJ | Negative | Negative | Positive | 46, XY |
P11029-6284-iPSpolyRoksmB | Negative | Negative | Positive | 46, XX |
P11030~5684~iPSpolyRoksmA | Negative | Negative | Positive | 46, XX, t(1;16)† |
P11031~5204~iPSpolyRoksmC | Negative | Negative | Positive | 46, XY |
-
*Tested in originating fibroblast cell lines.
-
†As found in originating fibroblasts.
Table 3
Related to Figure 2. Single nucleotide polymorphisms in ANK2, SCN5A, KCNQ1, CACNA1C, CALM1, KCNE2, KCNH2 and KCNJ5 as being significantly imbalanced (p<0.05) between low-S vs. high-S groups. …
Gene | Rs number | Anticipated MAF | Observed MAF Low-S | Observed MAF High-S | p value |
|---|---|---|---|---|---|
ANK2 | rs17045935 | 6.8% | 0% | 22.2% | 0.03 |
rs62314901 | 40% | 56.25% | 27.8% | 0.037 | |
rs17676256 | 10.2% | 12.5% | 38.9% | 0.03 | |
rs967099 | 38.1% | 25% | 44.4% | 0.027 | |
rs4834321 | 46.9% | 37.5% | 55.6% | 0.02 | |
rs35308370 | 48.3% | 25% | 66.7% | 0.025 | |
rs931838 | 42.5% | 31.25% | 66.7% | 0.048 | |
SCN5A | rs7375123 | 19.2% | 43.75% | 11.2% | 0.007 |
rs12491987 | 7.6% | 0% | 22.2% | 0.03 | |
rs9871385 | 34.1% | 6.25% | 33.3% | 0.02 | |
rs9818148 | 19.2% | 56.25% | 16.7% | 0.049 | |
KCNQ1 | rs4255520 | 15.5% | 18.75% | 0% | 0.043 |
rs151288 | 32.7% | 0% | 22.2% | 0.031 | |
rs718579 rs11022996 | 39.8% 41.2% | 12.5% | 27.8% | 0.034 | |
rs151212 | 40.8% | 50% | 33.3% | 0.05 | |
CACNA1C | rs3794299 | 12.4% | 18.75% | 0% | 0.043 |
rs4765661 rs2238018 | 15.8% 17.3% | 31.25% | 5.6% | 0.027 | |
CALM1 | rs3814843 | 1.8% | 0% | 22.2% | 0.031 |
rs2300502 | 7.5% | 6.25% | 33.3% | 0.023 | |
KCNE2 | rs28409368 | 30.8% | 18.75% | 38.9% | 0.0239 |
KCNH2 | rs2072411 | 37.5% | 18.75% | 44.4% | 0.027 |
rs3778873 | 15.8% | 6.2% | 44.4% | 0.02 | |
KCNJ5 | rs7924416 | 24.6% | 0% | 27.8% | 0.013 |
Table 4
Related to Figure 4. AP parameters of low-S and high-S iPSC-derived cardiomyocytes at baseline control condition. AP data are mean ± SE. APD50/APD90, AP duration measured at 50% or 90% …
Low-S (11009) (n=7) | High-S (11029) (n=5) | |
|---|---|---|
Firing Frequency (mV) | 1.26 ± 0.29 | 1.03 ± 0.28 |
Amplitude (mV) | 88.3 ± 3.6 | 85.1 ± 4.7 |
Upstroke velocity (mV/ms) | 11.7 ± 1.0 | 14.0 ± 2.0 |
Decay velocity (mV/ms) | −7.2 ± 1.4 | −11.1 ± 3.5 |
APD50 (ms) | 306.4 ± 57.9 | 299.4 ± 22.0 |
APD90 (ms) | 341.8 ± 58.6 | 338.4 ± 21.4 |
MDP (mV) | −69.4 ± 3.2 | −68.0 ± 1.3 |
Table 5
Related to Figure 5. Generation of new groups associating one high-S and one low-S line of the same gender (six groups for males and 16 groups for females) in order to neutralize gender-specific …
Male groups | Female groups |
|---|---|
P11019 (high) + P11028 (low) | P11008 (high) + P11030 (low) |
P11019 (high) + P11026 (low) | P11015 (high) + P11030 (low) |
P11019 (high) + P11020 (low) | P11013 (high) + P11030 (low) |
P11019 (high) + P11009 (low) | P11029 (high) + P11030 (low) |
P11019 (high) + P11031 (low) | P11018 (high) + P11030 (low) |
P11019 (high) + P11007 (low) | P11024 (high) + P11030 (low) |
P11023 (high) + P11030 (low) | |
P11021 (high) + P11030 (low) | |
P11008 (high) + P11014 (low) | |
P11015 (high) + P11014 (low) | |
P11013 (high) + P11014 (low) | |
P11029 (high) + P11014 (low) | |
P11018 (high) + P11014 (low) | |
P11024 (high) + P11014 (low) | |
P11023 (high) + P11014 (low) | |
P11021 (high) + P11014 (low) |
