Improving Generation of Cardiac Organoids from Human Pluripotent Stem Cells Using the Aurora Kinase Inhibitor ZM447439

Abstract

Drug-induced cardiotoxicity reduces the success rates of drug development. Thus, the limitations of current evaluation methods must be addressed. Human cardiac organoids (hCOs) derived from induced pluripotent stem cells (hiPSCs) are useful as an advanced drug-testing model; they demonstrate similar electrophysiological functionality and drug reactivity as the heart. How-ever, similar to other organoid models, they have immature characteristics compared to adult hearts, and exhibit batch-to-batch variation. As the cell cycle is important for the mesodermal differentiation of stem cells, we examined the effect of ZM447439, an aurora kinase inhibitor that regulates the cell cycle, on cardiogenic differentiation. We determined the optimal concentration and timing of ZM447439 for the differentiation of hCOs from hiPSCs and developed a novel protocol for efficiently and reproducibly generating beating hCOs with improved electrophysiological functionality, contractility, and yield. We validated their maturity through electro-physiological- and image-based functional assays and gene profiling with next-generation sequencing, and then applied these cells to multi-electrode array platforms to monitor the cardio-toxicity of drugs related to cardiac arrhythmia; the results confirmed the drug reactivity of hCOs. These findings may enable determination of the regulatory mechanism of cell cycles underlying the generation of iPSC-derived hCOs, providing a valuable drug testing platform.

Cancer organoids: A platform in basic and translational research

“… 139 Similarly, cardiac organoids were used to demonstrate that hypoxic cardiac injury aggravates the cardiotoxicity of doxorubicin. 141 Cardiac organoids from mouse embryonic stem cell-derived embryoid bodies 142 and human iPSCs have also been used for drug testing, 143 as have iPSC-derived kidney organoids for nephrotoxicity screening. 144 These organoids provide patient-specific models for investigating the toxicity of anticancer drugs in personalized medicine.…”

Revolutionizing Disease Modeling: The Emergence of Organoids in Cellular Systems

“…This can help researchers understand the underlying causes of congenital defects and can also be used to test the effectiveness of new treatments for these conditions. Human cardiac organoid shows similar constructs to in vivo organ, modeling cardiovascular disorders, such as heart failure, genetic cardiac diseases, and arrhythmia, providing a valuable drug testing platform [ 228 , 229 , 230 , 231 ]. Additionally, organoids were used to study immune tissue structures and functions, such as gut organoids for bowel disease [ 232 ], thymus organoids to induce donor-specific immune tolerance to allografts [ 233 ], lymph node organoids for regulating antibody production [ 234 ], and intestinal organoids to study Crohn’s disease [ 235 ].…”

A review of protocols for engineering human cardiac organoids

“…Similarly, Su-Jin Lee used low-adhesion 6-well plates and replaced IWP2 with IWP4. LAAP and ZM447439 were added to RB-medium, with ZM447439 increasing the expression of non-CMs and CMs during hCO generation [ 42 ]. Seul-Gi Lee modified the protocol by introducing ECM components to support the organoid structure geometrically, to enable organoid scale-up in a 60 cm dish [ 43 ].…”