Functional regulation of YAP mechanosensitive transcriptional coactivator by Focused Low-Intensity Pulsed Ultrasound (FLIPUS) enhances proliferation of murine mesenchymal precursors

Regina Puts; Paul Rikeit; Karen Ruschke; Petra Knaus; Sophie Schreivogel; Kay Raum

doi:10.1371/journal.pone.0206041

Functional regulation of YAP mechanosensitive transcriptional coactivator by Focused Low-Intensity Pulsed Ultrasound (FLIPUS) enhances proliferation of murine mesenchymal precursors

PLoS One. 2018 Oct 26;13(10):e0206041. doi: 10.1371/journal.pone.0206041. eCollection 2018.

Authors

Regina Puts^{1

2}, Paul Rikeit^{2

3}, Karen Ruschke³, Petra Knaus^{1

2

3}, Sophie Schreivogel^{2

4}, Kay Raum^{1

2}

Affiliations

¹ Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Charité-Berlin University of Medicine, Berlin, Germany.
² Berlin-Brandenburg School for Regenerative Therapies (BSRT), Charité-Berlin University of Medicine, Berlin, Germany.
³ Institute of Chemistry and Biochemistry, Free University of Berlin, Berlin, Germany.
⁴ Julius Wolff Institute, Charité-Berlin University of Medicine, Berlin, Germany.

Abstract

Yes-associated protein (YAP) acts as a mechanotransducer in determining the cell fate of murine C2C12 mesenchymal precursors as investigated after stimulation with ultrasound. We applied Focused Low-Intensity Pulsed Ultrasound (FLIPUS) at a sound frequency of 3.6 MHz, 100 Hz pulse repetition frequency (PRF), 27.8% duty cycle (DC), and 44.5 mW/cm2 acoustic intensity ISATA for 5 minutes and evaluated early cellular responses. FLIPUS decreased the level of phosphorylated YAP on Serine 127, leading to higher levels of active YAP in the nucleus. This in turn enhanced the expression of YAP-target genes associated with actin nucleation and stabilization, cytokinesis, and cell cycle progression. FLIPUS enhanced proliferation of C2C12 cells, whereas silencing of YAP expression abolished the beneficial effects of ultrasound. The expression of the transcription factor MyoD, defining cellular myogenic differentiation, was inhibited by mechanical stimulation. This study shows that ultrasound exposure regulates YAP functioning, which in turn improves the cell proliferative potential, critical for tissue regeneration process.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Actins / metabolism
Adaptor Proteins, Signal Transducing / genetics
Adaptor Proteins, Signal Transducing / metabolism*
Animals
Cell Cycle Proteins
Cell Line
Cell Nucleus / metabolism
Cell Proliferation
Cytoskeleton / metabolism
Gene Expression Regulation
Mechanotransduction, Cellular*
Mesenchymal Stem Cells / cytology*
Mesenchymal Stem Cells / metabolism*
Mice
Models, Biological
Phosphoproteins / genetics
Phosphoproteins / metabolism*
Phosphorylation
Phosphoserine / metabolism
RNA, Messenger / genetics
RNA, Messenger / metabolism
Trans-Activators / genetics
Trans-Activators / metabolism*
Ultrasonic Waves*
YAP-Signaling Proteins

Substances

Actins
Adaptor Proteins, Signal Transducing
Cell Cycle Proteins
Phosphoproteins
RNA, Messenger
Trans-Activators
YAP-Signaling Proteins
Yap1 protein, mouse
Phosphoserine

Grants and funding

This work has been supported by the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG, http://www.dfg.de/) grant Ra1380/8-1 (R.P. and K.R.); by the DFG grant FOR2165 “Regeneration in Aged Individuals” (P.R. and P.K.); by the DFG through the Berlin-Brandenburg School for Regenerative Therapies GSC 203 (fellowships to R.P. and P.R.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.