Oxylipin Signaling in Congenital Heart Disease

This project documents a highly faithful in vitro human heart organoid model developed to recapitulate early human cardiac development and to enable discovery and translational studies of cardiovascular disease, including congenital heart defects. The Series GSE153185 reports generation of human heart organoids (hHOs) from human pluripotent stem cells (hPSCs) using a fully defined, two-step manipulation of Wnt signaling with chemical inhibitors and growth factors. The protocol is described as efficient, scalable, reproducible, and compatible with high-throughput approaches, and organoids were derived from multiple independent hPSC lines with similar efficiency. hHOs begin spontaneous beating around day six and reach approximately 1 mm in diameter by day 15, developing sophisticated, interconnected internal chambers. Confocal marker analysis demonstrates the presence of all major cardiac lineages within the organoids: cardiomyocytes (TNNT2+), epicardial cells (WT1+, TJP+), cardiac fibroblasts (THY1+, VIM+), endothelial cells (PECAM1+), and endocardial cells (NFATC1+). Morphologically, the organoids establish well-defined epicardial and adjacent myocardial regions, a distinct vascular plexus, and endocardial-lined microchambers, indicating that higher-order tissue interactions and three-dimensional physical and topographical cues reminiscent of the human fetal heart are present in this system.

Comparative RNA-seq time-course analysis across hHOs, monolayer-differentiated iPSCs, and fetal human hearts indicates that hHOs better recapitulate human fetal heart tissue development than previously described differentiation protocols, supporting their value as a model for studying developmental gene expression dynamics and tissue complexity. The dataset includes RNA-seq performed on organoids sampled at multiple differentiation time points as well as monolayer cardiomyocyte controls, enabling global expression profiling to assess developmental commitment and lineage specification over time. Contributors listed for the series include Lewis-Israeli Y, Gabalski M, Ball KA, Wasserman AH, and Aguirre A, and the submission references NIH grant support, including Grant IDs K01 HL135464 (Lipid signaling in cardiac development and disease) and R01 HL151505 (Oxylipin Signaling in Congenital Heart Disease) affiliated with Michigan State University and investigator Aitor Aguirre.

The resource provides raw data availability in SRA and processed files on the GEO Series record, along with supplementary spreadsheets describing cardiac differentiation and organoid samples. Overall, this work establishes a reproducible, multi-lineage, three-dimensional human heart organoid platform that closely models fetal cardiac morphology and cellular composition, offering a powerful tool for mechanistic studies of heart development, congenital heart defects, and related signaling pathways such as lipid and oxylipin signaling, as well as for high-throughput translational applications.