STRIP2 Is Indispensable for the Onset of Embryonic Stem Cell Differentiation

Davood Sabour; Sureshkumar Perumal Srinivasan; Susan Rohani; Vilas Wagh; John Antonydas Gaspar; Darius Panek; Mostafa Abootorabi Ardestani; Michael Xavier Doss; Nicole Riet; Hinrich Abken; Jürgen Hescheler; Symeon Papadopoulos; Agapios Sachinidis

doi:10.1016/j.omtm.2017.04.001

STRIP2 Is Indispensable for the Onset of Embryonic Stem Cell Differentiation

Mol Ther Methods Clin Dev. 2017 Apr 14:5:116-129. doi: 10.1016/j.omtm.2017.04.001. eCollection 2017 Jun 16.

Affiliations

¹ Institute of Neurophysiology and Center for Molecular Medicine Cologne (CMMC), University of Cologne (UKK), Robert-Koch-Strasse 39, 50931 Cologne, Germany.
² Institute of Vegetative Physiology, Center of Physiology and Pathophysiology, University of Cologne, Robert-Koch-Strasse 39, 50931 Cologne, Germany.
³ Department I for Internal Medicine, Center for Molecular Medicine Cologne, Robert-Koch-Strasse 21, University of Cologne, and University Hospital Cologne, 50931 Cologne, Germany.

Abstract

The role of striatin interacting protein 2 (Strip2) in differentiation of embryonic stem cells (ESCs) is still under debate. Strip2-silenced murine (KD) ESCs were differentiated for 4, 8, 12, and 16 days. We show that Strip2 is distributed in the perinucleus or nuclei of wild-type (WT) undifferentiated ESCs, but is localized in high-density nuclear bodies in differentiated cells. CellNet analysis of microarray gene expression data for the KD and scrambled control (SCR) embryoid bodies (EBs), as well as immunostainings of key pluripotent factors, demonstrated that differentiation of KD ESCs is repressed. This occurs even in 16-day-old EBs, which possessed a high tumorigenic potential. Correlated with very high expression levels of epigenetic regulator genes, Hat1 and Dnmt3, enzymatic activities of the histone acetyltransferase type B (Hat1) and DNA (cytosine-5)-methyltransferase 3 beta (Dnmt3b) were higher in differentiated 16-day-old KD EBs than in SCR or WT EBs. The expression levels of let-7, 290, and 302 microRNA families were opposed in KD ESCs, while KD EBs had levels comparable to WT and SCR ESCs during differentiation. Strip2 is critical for the regular differentiation of ESCs. Moreover, Strip2 deficient ESCs showed a dysregulation of epigenetic regulators and microRNAs regulating pluripotency.

Keywords: Strip2; embryonic stem cells; epigenetic factors; microRNAs; ncRNAs; pluripotency; shRNA gene silencing; stem cell differentiation; striatin interacting protein 2.