S-nitrosoglutathione reductase (GSNOR) deficiency accelerates cardiomyocyte differentiation of induced pluripotent stem cells.

INTRODUCTION

Induced pluripotent stem cells (iPSCs) provide a model of cardiomyocyte (CM) maturation. Nitric oxide signaling promotes CM differentiation and maturation, although the mechanisms remain controversial.

AIM

The study tested the hypothesis that in the absence of S-nitrosoglutathione reductase (GSNOR), a denitrosylase regulating protein S-nitrosylation, the resultant increased S-nitrosylation accelerates the differentiation and maturation of iPSC-derived cardiomyocytes (CMs).

METHODS AND RESULTS

iPSCs derived from mice lacking GSNOR (iPSC) matured faster than wildtype iPSCs (iPSC) and demonstrated transient increases in expression of murine Snail Family Transcriptional Repressor 1 gene (), murine Snail Family Transcriptional Repressor 2 gene () and murine Twist Family BHLH Transcription Factor 1 gene (), transcription factors that promote epithelial-to-mesenchymal transition (EMT) and that are regulated by Glycogen Synthase Kinase 3 Beta (GSK3β). Murine Glycogen Synthase Kinase 3 Beta () gene exhibited much greater S-nitrosylation, but lower expression in iPSC. S-nitrosoglutathione (GSNO)-treated iPSC and human (h)iPSCs also demonstrated reduced expression of GSK3β. expression, a CM marker, was increased in iPSC upon directed differentiation toward CMs on Day 4, whereas murine Brachyury (), , and GATA Binding Protein () mRNA were decreased, compared to iPSC, suggesting that GSNOR deficiency promotes CM differentiation beginning immediately following cell adherence to the culture dish-transitioning from mesoderm to cardiac progenitor.

CONCLUSION

Together these findings suggest that increased S-nitrosylation of Gsk3β promotes CM differentiation and maturation from iPSCs. Manipulating the post-translational modification of GSK3β may provide an important translational target and offers new insight into understanding of CM differentiation from pluripotent stem cells.

ONE SENTENCE SUMMARY

Deficiency of GSNOR or addition of GSNO accelerates early differentiation and maturation of iPSC-cardiomyocytes.