Method Profile
Four computationally prioritized constituents are tested for direct toxicity and protection during 24-hour doxorubicin coexposure in human cardiac organoids.
Compare measured cardioprotective responses with network/docking predictions and identify active concentrations that avoid candidate toxicity.
Human cardiac organoids differentiated from commercially obtained iPSCs; the study does not specify independent donor or line replication.
24-hour doxorubicin concentration series followed by compound-alone and simultaneous compound/doxorubicin tests, with metabolic, secretory, protein and histological endpoints.
Injury conditions
Doxorubicin 0.1, 0.3, 1, 3, 10 and 30 μM for 24 hours; 10 μM selected for approximately 50% lower CCK-8 signal.
Candidate conditions
Quercetin, kaempferol, licochalcone A and naringenin; compound-alone 5–40 μM. Mechanistic series: quercetin 10–40 μM and kaempferol 5–20 μM; further histology/cytokines used quercetin 20–40 μM and kaempferol 10–20 μM.
Prioritization
Ingredient/heart-failure network intersection and MAPK14 docking/100-ns simulation preceded experimental testing; no direct biochemical target-engagement assay.
Compound-only toxicity and simultaneous doxorubicin coexposure are evaluated separately. Favorable docking scores occurred alongside unsuccessful candidates and a nonsignificant initial LDH increase. Reduced p38/ERK phosphorylation is associated with protection, without direct causal target confirmation.
Four-candidate comparison after computational ranking, with concentration selection and orthogonal injury measurements.
Figures 5–6 n = 6; Figures 7–8 n = 3. These assay counts are not reported as independent donor counts.
Untreated, drug-alone, doxorubicin-alone and simultaneous candidate/doxorubicin conditions.
Quercetin/kaempferol improved viability and several injury readouts; licochalcone A/naringenin failed protective concentration testing. Kaempferol and licochalcone A at 40 μM reduced viability. Initial doxorubicin-induced LDH increase was nonsignificant. Protective conditions reduced p38/ERK phosphorylation, apoptosis/inflammatory and collagen-associated signals.
Four network-prioritized compounds were tested in human iPSC-derived cardiac organoids. Quercetin and kaempferol improved 24-hour doxorubicin injury readouts, while licochalcone A and naringenin failed to provide effective protection and high-dose kaempferol was cytotoxic.
