Brichta L, Hofmann Y, Hahnen E, Siebzehnrubl F A, Raschke H, Blumcke I, Eyupoglu I Y, Wirth B
Institute of Human Genetics, University of Bonn, Wilhelmstrasse 31, 53111 Bonn, Germany.
Hum Mol Genet. 2003 Oct 1;12(19):2481-9. doi: 10.1093/hmg/ddg256. Epub 2003 Jul 29.
Proximal spinal muscular atrophy (SMA) is a common neuromuscular disorder causing infant death in half of all patients. Homozygous absence of the survival motor neuron gene (SMN1) is the primary cause of SMA, while SMA severity is mainly determined by the number of SMN2 copies. One SMN2 copy produces only about 10% of full-length protein identical to SMN1, whereas the majority of SMN2 transcripts is aberrantly spliced due to a silent mutation within an exonic splicing enhancer in exon 7. However, correct splicing can be restored by over-expression of the SR-like splicing factor Htra2-beta 1. We show that in fibroblast cultures derived from SMA patients treated with therapeutic doses (0.5-500 microM) of valproic acid (VPA), the level of full-length SMN2 mRNA/protein increased 2- to 4-fold. Importantly, this up-regulation of SMN could be most likely attributed to increased levels of Htra2-beta 1 which facilitates the correct splicing of SMN2 RNA as well as to an SMN gene transcription activation. Especially at low VPA concentrations, the restored SMN level depended on the number of SMN2 copies. Moreover, VPA was able to increase SMN protein levels through transcription activation in organotypic hippocampal brain slices from rats. Finally, VPA also increased the expression of further SR proteins, which may have important implications for other disorders affected by alternative splicing. Since VPA is a drug highly successfully used in long-term epilepsy therapy, our findings open the exciting perspective for a first causal therapy of an inherited disease by elevating the SMN2 transcription level and restoring its correct splicing.
近端脊髓性肌萎缩症(SMA)是一种常见的神经肌肉疾病,半数患者会因此在婴儿期死亡。生存运动神经元基因(SMN1)的纯合缺失是SMA的主要病因,而SMA的严重程度主要由SMN2基因拷贝数决定。一个SMN2基因拷贝仅能产生约10%与SMN1相同的全长蛋白,而由于外显子7中外显子剪接增强子内的沉默突变,大多数SMN2转录本会发生异常剪接。然而,通过过表达SR样剪接因子Htra2-β1可以恢复正确剪接。我们发现,在接受治疗剂量(0.5 - 500微摩尔)丙戊酸(VPA)治疗的SMA患者来源的成纤维细胞培养物中,全长SMN2 mRNA/蛋白水平增加了2至4倍。重要的是,SMN的这种上调很可能归因于Htra2-β1水平的增加,它促进了SMN2 RNA的正确剪接以及SMN基因转录激活。特别是在低VPA浓度下,恢复的SMN水平取决于SMN2基因拷贝数。此外,VPA能够通过激活大鼠海马脑片的转录来增加SMN蛋白水平。最后,VPA还增加了其他SR蛋白的表达,这可能对受可变剪接影响的其他疾病具有重要意义。由于VPA是一种在长期癫痫治疗中非常成功使用的药物,我们的发现为通过提高SMN2转录水平和恢复其正确剪接来首次对遗传性疾病进行因果治疗开辟了令人兴奋的前景。