Santoro Massimo, Piacentini Roberto, Perna Alessia, Pisano Eugenia, Severino Anna, Modoni Anna, Grassi Claudio, Silvestri Gabriella
IRCCS Fondazione Don Carlo Gnocchi, Milan, Italy.
Department of Neuroscience, Università Cattolica del Sacro Cuore; Fondazione Policlinico Universitario A. Gemelli IRCCS, Rome, Italy.
Neural Regen Res. 2020 Sep;15(9):1757-1766. doi: 10.4103/1673-5374.276336.
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 Ca-ATPase (SERCA) and α1S subunit of voltage-gated Ca channels (Ca1.1) is related to Ca 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 Ca release from RYR1 dependent on the electro-mechanical coupling between RYR1 and Ca1.1. Interestingly, immunoblotting studies showed that RES treatment was associated with a reduction in the levels of CUGBP Elav-like family member 1, while RYR1, Ca1.1 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.
1型强直性肌营养不良（DM1）是一种剪接病，与核转录RNA结合因子被DMPK前体mRNA的非编码CUG重复序列隔离导致的多个基因错配剪接有关。兰尼碱受体1（RYR1）、肌浆网/内质网Ca-ATP酶（SERCA）和电压门控钙通道α1S亚基（Ca1.1）的失调与钙稳态及兴奋-收缩偶联受损有关。虽然目前尚无针对DM1的药物治疗方法，但异常剪接校正仍是该病的一个主要治疗靶点。白藜芦醇（RES，3,5,4'-三羟基反式芪）因其能够直接结合DNA和RNA影响基因表达及可变剪接，是一种有前景的DM1治疗药物工具。在此，我们在一项初步研究中分析了RES对DM1肌管的治疗效果，该研究包括来自两名DM1患者和两名健康对照的培养肌管。我们的结果表明，RES治疗纠正了RYR1的异常剪接，这一事件似乎与依赖于RYR1和Ca1.1之间机电偶联的去极化诱导的RYR1钙释放恢复有关。有趣的是，免疫印迹研究表明，RES治疗与CUGBP Elav样家族成员1水平的降低有关，而RYR1、Ca1.1和SERCA1蛋白水平未发生变化。最后，RES治疗在核仁病灶数量或肌肉盲样剪接调节因子1的隔离方面均未引起任何重大变化。总体而言，这项初步研究的结果支持将RES作为一种有吸引力的化合物用于未来DM1的临床试验。2014年5月20日，意大利罗马IRCCS圣心天主教大学综合医院基金会伦理委员会（rs9879/14）批准了该研究。
