Cyclooxygenases Inhibitors Efficiently Induce Cardiomyogenesis in Human Pluripotent Stem Cells
Authors: Jürgen Hescheler
Hinrich Abken
Nicole Riet
Erastus Nembo
Harshal Nemade
Umesh Chaudhari
Aviseka Acharya
Symeon Papadopoulos
Filomain Nguemo
Agapios Sachinidis
Journal: Cells (2020)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: Cardiomyocytes; Cyclooxygenase pathway; Differentiation; Pluripotent stem cells; Small molecules
Affiliations: Institute of Neurophysiology, Faculty of Medicine, University of Cologne, Robert-Koch-Str. 39, 50931 Cologne, Germany., Dept I Internal Medicine and Center for Molecular Medicine Cologne (CMMC), University of Cologne (UKK), Robert-Koch-Str. 21, 50931 Cologne, Germany., Regensburg Centre for Interventional Immunology (RCI), Dept. Genetic Immunotherapy, University Hospital Regensburg, 93053 Regensburg, Germany., Center for Molecular Medicine Cologne (CMMC), University of Cologne, Robert-Koch-Str. 21, 50931 Cologne, Germany.

Abstract:  Application of human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) is limited by the challenges in their efficient differentiation. Recently, the Wingless (Wnt) signaling pathway has emerged as the key regulator of cardiomyogenesis. In this study, we evaluated the effects of cyclooxygenase inhibitors on cardiac differentiation of hPSCs. Cardiac differentiation was performed by adherent monolayer based method using 4 hPSC lines (HES3, H9, IMR90, and ES4SKIN). The efficiency of cardiac differentiation was evaluated by flow cytometry and RT-qPCR. Generated hPSC-CMs were characterised using immunocytochemistry, electrophysiology, electron microscopy, and calcium transient measurements. Our data show that the COX inhibitors Sulindac and Diclofenac in combination with CHIR99021 (GSK-3 inhibitor) efficiently induce cardiac differentiation of hPSCs. In addition, inhibition of COX using siRNAs targeted towards COX-1 and/or COX-2 showed that inhibition of COX-2 alone or COX-1 and COX-2 in combination induce cardiomyogenesis in hPSCs within 12 days. Using IMR90-Wnt reporter line, we showed that inhibition of COX-2 led to downregulation of Wnt signalling activity in hPSCs. In conclusion, this study demonstrates that COX inhibition efficiently induced cardiogenesis via modulation of COX and Wnt pathway and the generated cardiomyocytes express cardiac-specific structural markers as well as exhibit typical calcium transients and action potentials. These cardiomyocytes also responded to cardiotoxicants and can be relevant as an in vitro cardiotoxicity screening model. 

Retracted: No

PubPeer comments:
 Figure 5a appears to have two images which overlap. *** ![file] , 
 Thank you for pointing out this mistake, we will rectify it immediately. , 
 Dear Actinopolyspora Biskrensis, Thank you very much for notifying us about the mistake in Figure 5a in our paper. Justification: For the WNT reporter assay, I have taken 6 photos from each well for each sample. While making the final version of the figures for submission I think I used two figures from Sulindac group which caused this duplication mistake. Update: I have corrected the Figure 5a now, and we (Prof. Sachinidis and I) will contact the publisher to get this mistake corrected as soon as possible. Once again thank you very much. Regards, Harshal Nemade. ![file] , 
 Published: 24 September 2020 The authors wish to make the following corrections to this paper [1]: For the WNT reporter assay, the authors have taken six photos from each well for each sample. While making the final version of the figures for submission, the authors used two figures from the Sulindac group, which caused a duplication mistake in Figure 5a. The corrected Figure 5a is shown as follows: ![file] ,