Fine-tuning cardiac differentiation with Wnt signaling and tranilast

Pluripotent stem cells can differentiate into various cell types under the influence of signaling pathways such as BMP4, Nodal, and Wnt/β-catenin. These pathways can be pharmacologically modulated to achieve in vitro differentiation into specific cell types. However, variations in cell line responses and the complexity of signaling interactions pose challenges in achieving consistent and efficient differentiation, which is critical for practical applications in medical research and therapy.

A recent study leverages advanced single-cell analytics and computational modeling, focusing on how controlled Wnt signaling influences the differentiation of human induced pluripotent stem cells (hiPSCs).

The researchers hypothesized that by gradually escalating the doses of a Wnt agonist, CHIR-99021, they could map out how different levels of Wnt activity influenced the fate of hiPSCs as they transitioned away from pluripotency.

They tracked how cells responded to these changes at a single-cell level using RNA sequencing techniques. Interestingly, they observed that different doses of CHIR-99021 led to the activation of distinct sets of genes, pushing the cells toward different developmental paths.

Analysis of gene expression associated with different Wnt doses allowed for hierarchical clustering of gene sets related to Wnt signaling pathways. A multiple regression model was then developed to predict cardiomyocyte differentiation efficiency based on these gene expression profiles.

Using the Connectivity Map database, the study identified tranilast—a drug previously used for treating allergies—as a small molecule that enhances cardiac differentiation. Tranilast was shown to synergistically promote cardiac lineage differentiation both in vitro and in vivo.

Experiments indicated that tranilast, combined with specific Wnt dosages, favored mesoderm over endoderm development, aligning with the desired outcomes for cardiac cell generation.

Physiological analysis of cardiac contractility, performed with the CardioExcyte 96 system, showed that tranilast-derived cardiomyocytes were physiologically similar to standard protocol derived cardiomyocytes.

Overall, this study demonstrates that controlled modulation of Wnt signaling can direct the differentiation of PSCs into specific cardiac cell types. Tranilast emerges as a potential enhancer of cardiac differentiation, suggesting its utility in improving the yield and quality of differentiated cells for therapeutic applications.

Find the original article here: https://www.cell.com/developmental-cell/abstract/S1534-5807(24)00039-X

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