中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (9): 1757-1766.doi: 10.4103/1673-5374.276336

• 原著:退行性病与再生 • 上一篇    下一篇

白藜芦醇可纠正强直性肌营养不良1型肌管中RYR1的异常剪接

  

  • 出版日期:2020-09-08 发布日期:2020-09-17

Resveratrol corrects aberrant splicing of RYR1 pre-mRNA and Ca2+ signal in myotonic dystrophy type 1 myotubes

Massimo Santoro1, * , Roberto Piacentini2, 4, * , Alessia Perna2 , Eugenia Pisano3, 4, Anna Severino3 , Anna Modoni4 , Claudio Grassi2, 4, Gabriella Silvestri2, 4    

  1. 1 IRCCS Fondazione Don Carlo Gnocchi, Milan, Italy 2 Department of Neuroscience, Università Cattolica del Sacro Cuore, Rome, Italy 3 Department of Cardiovascular and Thoracic Sciences, Università Cattolica del Sacro Cuore, Rome, Italy 4 Fondazione Policlinico Universitario A. Gemelli IRCCS, Rome, Italy
  • Online:2020-09-08 Published:2020-09-17
  • Contact: Roberto Piacentini, PhD, roberto.piacentini@unicatt.it; Massimo Santoro, PhD, masantoro@dongnocchi.it.
  • Supported by:
    This work was supported by grants from Università Cattolica and Italian Ministry of Scientific Research (grant number D1-2016; to GS).

摘要:

orcid: 0000-0003-4215-1643 (Roberto Piacentini) 

          0000-0002-3037-2775 (Massimo Santoro) 

          0000-0001-7253-1685 (Claudio Grassi)

Abstract: Myotonic dystrophy type 1 (DM1) is a spliceopathy related to the mis-splicing of several genes caused by sequestration of nuclear transcriptional RNA-binding factors from non-coding CUG repeats of DMPK pre-mRNAs. Dysregulation of ryanodine receptor 1 (RYR1), sarcoplasmatic/endoplasmatic Ca2+-ATPase (SERCA) and α1S subunit of voltage-gated Ca2+ channels (Cav1.1) is related to Ca2+ homeostasis and excitation-contraction coupling impairment. Though no pharmacological treatment for DM1 exists, aberrant splicing correction represents one major therapeutic target for this disease. Resveratrol (RES, 3,5,4′-trihydroxy-trans-stilbene) is a promising pharmacological tools for DM1 treatment for its ability to directly bind the DNA and RNA influencing gene expression and alternative splicing. Herein, we analyzed the therapeutic effects of RES in DM1 myotubes in a pilot study including cultured myotubes from two DM1 patients and two healthy controls. Our results indicated that RES treatment corrected the aberrant splicing of RYR1, and this event appeared associated with restoring of depolarization-induced Ca2+ release from RYR1 dependent on the electro-mechanical coupling between RYR1 and Cav1.1. Interestingly, immunoblotting studies showed that RES treatment was associated with a reduction in the levels of CUGBP Elav-like family member 1, while RYR1, Cav1.1 and SERCA1 protein levels were unchanged. Finally, RES treatment did not induce any major changes either in the amount of ribonuclear foci or sequestration of muscleblind-like splicing regulator 1. Overall, the results of this pilot study would support RES as an attractive compound for future clinical trials in DM1. Ethical approval was obtained from the Ethical Committee of IRCCS Fondazione Policlinico Universitario A. Gemelli, Rome, Italy (rs9879/14) on May 20, 2014.

Key words: alternative splicing, calcium homeostasis, CUG-BP1, foci, MBNL1, myotonic dystrophy type 1, myotubes, resveratrol