Repurposing of an inotropic drug dobutamine to enhance the production of human hematopoietic stem cells from human induced pluripotent stem cells.

Abstract

Background: Dobutamine hydrochloride (DH), a common inotropic drug used for heart failure, has recently been discovered to inhibit Yes-Associated Protein (YAP). YAP is a key component of the Hippo signaling pathway and plays a crucial role in the regulation of hematopoietic cell growth. The decrease in YAP activity has been shown to increase hematogenic differentiation and the generation of hematopoietic stem and progenitor cells (HSPCs) from human induced pluripotent stem cells (hiPSCs). Therefore, this study investigates the effect of DH on enhancing the hematopoietic differentiation of hiPSCs toward HSPCs.

Methods: This study used isogenic hiPSCs to study the effect of DH during various stages of their hematogenic differentiation using an in vitro culture system. The differentiating hiPSCs were cultured under specific conditions, including defined differentiation media composition and controlled oxygen tension throughout the differentiation process. The percentages of iPSC-derived HSPCs were assessed using flow cytometry to evaluate the expression of HSPC markers, including CD34⁺, CD43⁺, and CD45⁺/⁻. The HSPC production yield and the multilineage differentiation capacity of the resulting hiPSC-derived HSPCs were determined at the end of culture.

Results: The findings indicate that DH treatment significantly inhibits YAP activity and increases the hematogenic differentiation of hiPSCs and the yield of HSPCs at the end of culture. Specifically, inhibiting YAP activity with DH during the transition of hiPSCs from the hematoendothelial progenitor (HE) stage to the hematopoietic stage (endothelial to hematopoietic transition, EHT) proved to be the most effective in increasing HSPC production from hiPSCs.

Conclusions: This study highlights the potential of the inotropic drug DH as a novel agent to enhance hematogenic differentiation and improve the yield of hiPSC-derived hematopoietic stem and progenitor cells (HSPCs). DH was found to significantly inhibit YAP activity, which in turn promoted hematopoietic specification, particularly when administered during the critical endothelial-to-hematopoietic transition (EHT) stage. These findings suggest that repurposing DH could offer a valuable strategy to increase the efficiency of hiPSC-derived HSPC production, advancing its potential for therapeutic and clinical applications in regenerative medicine and hematopoietic cell therapies.