MBNL1 reverses the proliferation defect of skeletal muscle satellite cells in myotonic dystrophy type 1 by inhibiting autophagy via the mTOR pathway

Kai-Yi Song,Yue Ma,Qing-Zhe Hu,Rong-Rong Zhang,Xiu-Ming Guo,Lei Zhang,Ying-Chao Huo,Gang Zhang,Si-Yuan Huang,Hui-Qi Wang,Jin-Zhou Feng,Xin-Yue Qin

doi:10.1038/s41419-020-02756-8

Abstract

Skeletal muscle atrophy is one of the clinical symptoms of myotonic dystrophy type 1 (DM1). A decline in skeletal muscle regeneration is an important contributor to muscle atrophy. Skeletal muscle satellite cells (SSCs) drive skeletal muscle regeneration. Increased autophagy can reduce the proliferative capacity of SSCs, which plays an important role in the early regeneration of damaged skeletal muscle in DM1. Discovering new ways to restore SSC proliferation may aid in the identification of new therapeutic targets for the treatment of skeletal muscle atrophy in DM1. In the pathogenesis of DM1, muscleblind-like 1 (MBNL1) protein is generally considered to form nuclear RNA foci and disturb the RNA-splicing function. However, the role of MBNL1 in SSC proliferation in DM1 has not been reported. In this study, we obtained SSCs differentiated from normal DM1-04-induced pluripotent stem cells (iPSCs), DM1-03 iPSCs, and DM1-13-3 iPSCs edited by transcription activator-like (TAL) effector nucleases (TALENs) targeting CTG repeats, and primary SSCs to study the pathogenesis of DM1. DM1 SSC lines and primary SSCs showed decreased MBNL1 expression and elevated autophagy levels. However, DM1 SSCs edited by TALENs showed increased cytoplasmic distribution of MBNL1, reduced levels of autophagy, increased levels of phosphorylated mammalian target of rapamycin (mTOR), and improved proliferation rates. In addition, we confirmed that after MBNL1 overexpression, the proliferative capability of DM1 SSCs and the level of phosphorylated mTOR were enhanced, while the autophagy levels were decreased. Our data also demonstrated that the proliferative capability of DM1 SSCs was enhanced after autophagy was inhibited by overexpressing mTOR. Finally, treatment with rapamycin (an mTOR inhibitor) was shown to abolish the increased proliferation capability of DM1 SSCs due to MBNL1 overexpression. Taken together, these data suggest that MBNL1 reverses the proliferation defect of SSCs in DM1 by inhibiting autophagy via the mTOR pathway.

Highlights

Myotonic dystrophy type 1 (DM1) is a neuromuscular disease caused by expanded CTG repeats in the myotonic dystrophy protein kinase (DMPK) gene[1,2]
We have developed a method for the directional insertion of polyA signals (PASs) upstream of the DMPK CTG repeats in DM1-03 induced pluripotent stem cells (iPSCs), which eliminates nuclear RNA foci and reverses the phenotype of muscleblind-like 1 (MBNL1)
There were no differences in morphology among the three iPSC lines (Fig. 1a), all of which successfully differentiated into Skeletal muscle satellite cells (SSCs) (DM1-04, DM1-03, and DM1-13-3) and expressed the cell markers Pax[7] and MyoD (Fig. 1b)

Summary

Introduction

Myotonic dystrophy type 1 (DM1) is a neuromuscular disease caused by expanded CTG repeats in the myotonic dystrophy protein kinase (DMPK) gene[1,2]. DMPK gene is transcribed into CUGn-containing RNA, which forms nuclear RNA foci and disturbs RNAbinding proteins, resulting in reduced levels of MBNL1 and increased levels of CUG-binding protein 1 (CUGBP1)[3]. Muscle atrophy is one of the core symptoms of DM11,4, with decreased skeletal muscle regeneration as an important contributor[5]. New methods of enhancing the impaired regeneration of skeletal muscle are essential to improve skeletal muscle atrophy and the quality of life of DM1 patients. Skeletal muscle satellite cells (SSCs) are the source of skeletal muscle regeneration.

Methods

Results

Conclusion