Self-assembling human heart organoids for the modeling of cardiac development and congenital heart disease

Abstract

Congenital heart defects constitute the most common human birth defect, however understanding of how these disorders originate is limited by our ability to model the human heart accurately in vitro. Here we report a method to generate developmentally relevant human heart organoids by self-assembly using human pluripotent stem cells. Our procedure is fully defined, efficient, reproducible, and compatible with high-content approaches. Organoids are generated through a three-step Wnt signaling modulation strategy using chemical inhibitors and growth factors. Heart organoids are comparable to age-matched human fetal cardiac tissues at the transcriptomic, structural, and cellular level. They develop sophisticated internal chambers with well-organized multi-lineage cardiac cell types, recapitulate heart field formation and atrioventricular specification, develop a complex vasculature, and exhibit robust functional activity. We also show that our organoid platform can recreate complex metabolic disorders associated with congenital heart defects, as demonstrated by an in vitro model of pregestational diabetes-induced congenital heart defects.

Modeling Human Heart Development and Congenital Defects Using Organoids: How Close Are We?

“…These ECs expressed the EndoEC markers NFATC1 and NPR3 , and their transcriptomic profiles were comparable with human umbilical vein endothelial cells (HUVECs) and human cardiac microvascular endothelial cells (HCMECs). Consistently, Lewis-Israeli et al also identified CMs, ECs, and Fbs in their organoids [ 20 ]. We also developed heart organoids with the three major cardiac cell types [ 28 ].…”

“…However, lumen formation does not seem to rely on the EndoECs, as the lumen can still form when the EndoECs developed on the outer organoid surface after VEGF treatment [ 27 ]. Lewis-Israeli et al and our study also generated organoids with chamber-like structures, and these chambers also developed independently from the EndoECs [ 20 , 28 ]. As chamber formation in the current organoids does not go through the same process as in vivo heart lumen development, there is limited value in modeling human heart lumen formation under normal and diseased conditions using organoid cultures.…”

“…They identified NKX2-5, PDGFRA , and EOMES expression in the FHF progenitors and ISL1 , MEF2C , and TBX18 expression in SHF cells ( Figure 1 B). These progenitors were described to respectively develop into left and right ventricular CMs based on the expression of HAND1 and HAND2 [ 20 ].…”

“…Additionally, Lewis-Israeli et al induced proepicardial lineage in organoids by introducing CHIR (a WNT activator) at a relatively late differentiation stage (day 7). They found that a short period of CHIR treatment was sufficient to induce a layer of epicardium on the organoid outer surface, and the ratio of CMs to epicardial cells was like in vivo (60–65% cardiomyocytes:10–20% epicardial cells) [ 20 ].…”

Human Brain-Based Models Provide a Powerful Tool for the Advancement of Parkinson’s Disease Research and Therapeutic Development

“…The advent of organoids is considered a major breakthrough in stem cell research and has enabled advancements in the applications of human iPSC for disease modelling. Human heart organoids provide an in vitro model of cardiac development and congenital heart disease ( Lewis-Israeli et al, 2021 ), inner ear organoids have been generated for modelling deafness ( Romano et al, 2021 ), airway epithelial cell organoids are frequently used in lung research ( Eenjes et al, 2021 ), intestinal organoids enable in vitro investigations of gut pathology ( Puschhof et al, 2021 ), and it is even possible to derive sensorimotor organoids capable of forming neuromuscular junctions ( Pereira et al, 2021 ).…”