Ppp1r1b-lncRNA inhibits PRC2 at myogenic regulatory genes to promote cardiac and skeletal muscle development in mouse and human

Xuedong Kang; Yan Zhao; Glen Van Arsdell; Stanley F Nelson; Marlin Touma

doi:10.1261/rna.073692.119

Ppp1r1b-lncRNA inhibits PRC2 at myogenic regulatory genes to promote cardiac and skeletal muscle development in mouse and human

RNA. 2020 Apr;26(4):481-491. doi: 10.1261/rna.073692.119. Epub 2020 Jan 17.

Authors

Xuedong Kang^{1

2}, Yan Zhao^{1

2}, Glen Van Arsdell^{1

3}, Stanley F Nelson^{1

4

5

6}, Marlin Touma^{1

2

5

6

7

8

9}

Affiliations

¹ Department of Pediatrics, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California 90095, USA.
² Neonatal/Congenital Heart Laboratory, Cardiovascular Research Laboratory, University of California Los Angeles, Los Angeles, California 90095, USA.
³ Department of Cardiothoracic Surgery, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California 90095, USA.
⁴ Department of Neurology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California 90095, USA.
⁵ Department of Human Genetics, Institute of Precision Health, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California 90095, USA.
⁶ Institute of Precision Health, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California 90095, USA.
⁷ The Molecular Biology Institute, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California 90095, USA.
⁸ Children's Discovery and Innovation Institute, Department of Pediatrics, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California 90095, USA.
⁹ Eli and Edythe Broad Stem Cell Institute, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California 90095, USA.

Abstract

Long noncoding RNAs (lncRNAs) have emerged as critical epigenetic regulators and play important roles in cardiac development and congenital heart disease. In a previous study, we identified a novel lncRNA, Ppp1r1b, with expression highly correlated with myogenesis. However, the molecular mechanism that underlies Ppp1r1b-lncRNA function in myogenic regulation is unknown. By silencing Ppp1r1b-lncRNA, mouse C2C12 and human skeletal myoblasts failed to develop fully differentiated myotubes. Myogenic differentiation was also impaired in PPP1R1B-lncRNA deficient human-induced pluripotent stem cell-derived cardiomyocytes (hiPSCs-CMs). The expression of myogenic transcription factors, including MyoD, Myogenin, and Tbx5, as well as sarcomere proteins, was significantly suppressed in Ppp1r1b-lncRNA inhibited myoblast cells and neonatal mouse heart. Histone modification analysis revealed increased H3K27 tri-methylation at MyoD1 and Myogenin promoters in GapmeR treated C2C12 cells. Furthermore, Ppp1r1b-lncRNA was found to bind to Ezh2, and chromatin isolation by RNA purification (ChIRP) assay revealed enriched interaction of Ppp1r1b-lncRNA with Myod1 and Tbx5 promoters, suggesting that Ppp1r1b-lncRNA induces transcription of myogenic transcription factors by interacting with the polycomb repressive complex 2 (PRC2) at the chromatin interface. Correspondingly, the silencing of Ppp1r1b-lncRNA increased EZH2 binding at promoter regions of myogenic transcription factors. Therefore, our results suggest that Ppp1r1b-lncRNA promotes myogenic differentiation through competing for PRC2 binding with chromatin of myogenic master regulators during heart and skeletal muscle development.

Keywords: H3K27me3; epigenome; long noncoding RNA; myogenesis.

Publication types

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

MeSH terms

Animals
Cell Line
Dopamine and cAMP-Regulated Phosphoprotein 32 / genetics*
Dopamine and cAMP-Regulated Phosphoprotein 32 / metabolism
Enhancer of Zeste Homolog 2 Protein / genetics
Enhancer of Zeste Homolog 2 Protein / metabolism
Gene Expression Regulation, Developmental*
Gene Silencing
Histone Code
Humans
Mice
Mice, Inbred C57BL
Muscle Development*
Muscle, Skeletal / metabolism*
MyoD Protein / genetics
MyoD Protein / metabolism
Myocytes, Cardiac / metabolism*
Myogenin / genetics
Myogenin / metabolism
Polycomb Repressive Complex 2 / metabolism*
RNA, Long Noncoding / genetics*
RNA, Long Noncoding / metabolism
T-Box Domain Proteins / genetics
T-Box Domain Proteins / metabolism

Substances

Dopamine and cAMP-Regulated Phosphoprotein 32
MyoD Protein
MyoD1 myogenic differentiation protein
Myogenin
Ppp1r1b protein, mouse
RNA, Long Noncoding
T-Box Domain Proteins
T-box transcription factor 5
Enhancer of Zeste Homolog 2 Protein
Ezh2 protein, mouse
Polycomb Repressive Complex 2

Abstract

Publication types

MeSH terms

Substances

Grants and funding