Normalized downstroke time (A), decay time (B), and beating rate as assessed by beats per minute (C) in hiPSC-CM treated with BSA (control), TNF-α, IFN-γ, TNF-α plus mitoTempo, and IFN-γ plus mitoTempo. N=5-14. Data were analyzed by ordinary one-way ANOVA and post-hoc Tukey’s multiple comparison test. Bars represent group mean.

Oxygen consumption rate (OCR) in hiPSC-CM treated with cytokines and antioxidant agent NAC. (A) OCR trace of hiPSC-CM treated with 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 Panel 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.

Decay time (a measurement of cardiomyocyte relaxation) in hiPSC-CM after treatment with TNF-α and various 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, raltegravier, and TNF-α+ raltegravier. (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.

Decay time in 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+tenofovir. (B) Decay time after treatment with TNF-α, sildenafil citrate, and TNF-α+sildenafil citrate. (C) Decay time after treatment with TNF-α, PF-04447943, and TNF-α+PF-04447943. (D) Decay time after treatment with TNF-α, dapagliflozin, and TNF-α+dapagliflozin. (E) Decay time after treatment with INF-γ, dapagliflozin, and IFN-γ+dapagliflozin. (F) Downstroke time after treatment with TNF-α, dapagliflozin, and TNF-α+dapagliflozin. (G) 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.

Diastolic function of hiPSC-CM and angiogenic function of ECs after treatment with 10% serum from Group 1 HIV+ patients with DD. (A-C) Pooled normalized decay time (A), downstroke time (B) and beating rate (C) in hiPSC treated with 10% serum for 48 hours from group 1 HIV+ patients with DD. (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 HUVECs after treatment with serum from Group 1 patients for 20 hours. Data were analyzed by unpaired Student’s t test. Bars represent group mean.

Diastolic function of hiPSC-CMs and angiogenic function of ECs after treatment with serum from HIV+ patients with DD (Group 2). (A-B) Individual normalized decay time 3 hours (N=3-9)(A) or 24 hours (N=3-13)(B) after treatment of hiPSC-CM with 2% serum from patients. (C-D) Pooled decay time data from patients in panels A and B with 3 hour (C) and 24 hours (D) after treatment with serum. (E) Representative image of formed tube structure with cultured HUVECs treated with serum from HIV+ patients with DD. (F-I) Assessment of tube formation of HUVECs 20 hours 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 and I). Bars represent group mean.

Mitochondrial membrane potential as assessed by tetramethylrhodamine, ethyl ester (TMRE) (A) in hiPSC-CM treated with various cytokines. Concentration of cytokines used is as follows: CCL3 1nM, CCL4 300 pM, CCL5 10 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 1ng/ml. N=4, Data were analyzed by ordinary one-way ANOVA and post-hoc Tukey’s multiple comparison test. Bars represent group mean.

Cell viability (A) and cellular ROS levels (B) in hiPSC-CM exposed to concentrations ranging from between 3nM and 10μM of tenofovir (a nucleotide-analog reverse transcriptase inhibitor), darunavir (a protease inhibitor), raltegravir and elvitegravir (integrase inhibitors).

Representative images of formed tube structure with HUVECs treated with 1% serum from HIV+ patients with high or low MPR for 20 hours, or with 1% serum from the same patients for 20 hours.