Computational and in vitro validation of cardiogenic induction of quercetin on adipose‐derived mesenchymal stromal cells through the inhibition of Wnt and non‐Smad‐dependent TGF‐β pathways

Abstract Exploiting human mesenchymal stem cells (hMSCs) was proposed as a promising therapeutic approach for cardiovascular disease due to their capacity to differentiate into cardiac cells. Though modulation of the intracellular signaling pathways dominantly WNT/β catenin and transforming growth factor‐β (TGF‐β) have been reported to promote differentiation of hMSCs into cardiomyocytes in the prevailing literature, a safe and reproducible system for their clinical application has not yet turned into reality. In the present study, the molecular docking‐based strategy was first applied for evaluating the potency of some natural phenolic compounds in the modulation of Wnt and TGF‐β signaling pathways using a vital class of crystallographic protein structures of WNT signaling regulators such as Frizzled, Disheveled, GSK3‐β, β‐catenin, LRP 5/6 extracellular domain, Tankyrase and their variety of active pockets. Then, the impacts of plant‐derived chemical compounds on the regulation of the relevant signals for the differentiation of hMSCs into the definitive mesoderm lineage and cardiac progenitors were assessed in vitro. Data obtained revealed the synergistic activity of Wnt and TGF‐β superfamily to direct cardiac differentiation in human cardiogenesis by comparing cardiac gene expression in the presence and absence of the TGF‐β inhibitors. We found that the inhibitory effect of canonical Wnt/β‐catenin is sufficient to cause proper cardiomyocyte differentiation, but the TGF‐β pathway plays a vital role in enhancing the expression of the cardiomyocyte‐specific marker (cTnT). It was found that quercetin, a p38MAPK inhibitor with the high energy dock to the active pocket of Wnt receptors, promotes cardiac differentiation via the inhibition of both Wnt and non‐Smad TGF‐β pathways. Altogether, data presented here can contribute to the development of a feasible and efficient cardiac differentiation protocol as an “off‐the‐shelf” therapeutic source using novel natural agents for cardiac repair or regeneration.