ZNF746/PARIS overexpression induces cellular senescence through FoxO1/p21 axis activation in myoblasts

Ju-Hyeon Bae,Dongryeol Ryu,Jong-Sun Kang,Sang Chul Park,Hyebeen Kim,Yun-Il Lee,Hyeon-Ju Jeong,Young-Eun Leem,Sung Chun Cho

doi:10.1038/s41419-020-2552-7

Abstract

Various stresses, including oxidative stress, impair the proliferative capacity of muscle stem cells leading to declined muscle regeneration related to aging or muscle diseases. ZNF746 (PARIS) is originally identified as a substrate of E3 ligase Parkin and its accumulation is associated with Parkinson’s disease. In this study, we investigated the role of PARIS in myoblast function. PARIS is expressed in myoblasts and decreased during differentiation. PARIS overexpression decreased both proliferation and differentiation of myoblasts without inducing cell death, whereas PARIS depletion enhanced myoblast differentiation. Interestingly, high levels of PARIS in myoblasts or fibroblasts induced cellular senescence with alterations in gene expression associated with p53 signaling, inflammation, and response to oxidative stress. PARIS overexpression in myoblasts starkly enhanced oxidative stress and the treatment of an antioxidant Trolox attenuated the impaired proliferation caused by PARIS overexpression. FoxO1 and p53 proteins are elevated in PARIS-overexpressing cells leading to p21 induction and the depletion of FoxO1 or p53 reduced p21 levels induced by PARIS overexpression. Furthermore, both PARIS and FoxO1 were recruited to p21 promoter region and Trolox treatment attenuated FoxO1 recruitment. Taken together, PARIS upregulation causes oxidative stress-related FoxO1 and p53 activation leading to p21 induction and cellular senescence of myoblasts.

Highlights

Skeletal muscle is maintained by stem cells called satellite cells, and upon injury quiescent satellite cells are activated, proliferated, and differentiated to regenerate muscles[1,2,3]
PARIS overexpression attenuates myoblast differentiation To examine the role of PARIS in myoblast function, the expression of PARIS was examined during C2C12 myoblast differentiation
The protein level of myosin heavy chain (MHC) and Troponin T (TnT) was elevated in PARISdepleted cells compared with the control scrambled small interference RNAs (siRNAs)-expressing cells (Fig. 1j, k)

Summary

Introduction

Skeletal muscle is maintained by stem cells called satellite cells, and upon injury quiescent satellite cells are activated, proliferated, and differentiated to regenerate muscles[1,2,3]. The impaired regenerative capacity associated with muscle aging is linked with the reduced number and function of satellite cells due to failure to retain quiescent state and increased senescence[4,5,6]. Among diverse downstream targets of oxidative stress, FoxO transcription factors are recently proposed to play key roles in induction of cellular senescence[9,10,11]. FoxO transcription factors are originally identified as the Official journal of the Cell Death Differentiation Association. In response to the stimulation of transforming growth factor-β, FoxO proteins form a complex with activated Smad proteins to induce the expression of p21, leading to cell cycle arrest[15,16]. The diverse functions of FoxO proteins are dependent on the cellular contexts likely linked with distinct regulatory proteins

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Results

Conclusion