Lin Lan, Park Juw Won, Ramachandran Shyam, Zhang Yida, Tseng Yu-Ting, Shen Shihao, Waldvogel Henry J, Curtis Maurice A, Faull Richard L M, Troncoso Juan C, Pletnikova Olga, Ross Christopher A, Davidson Beverly L, Xing Yi
Department of Microbiology, Immunology, & Molecular Genetics, University of California Los Angeles, Los Angeles, CA, USA.
The Raymond G Perelman Center for Cellular and Molecular Therapy, The Children's Hospital of Philadelphia, PA, USA.
Hum Mol Genet. 2016 Aug 15;25(16):3454-3466. doi: 10.1093/hmg/ddw187. Epub 2016 Jul 4.
Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by a CAG expansion in the gene-encoding Huntingtin (HTT). Transcriptome dysregulation is a major feature of HD pathogenesis, as revealed by a large body of work on gene expression profiling of tissues from human HD patients and mouse models. These studies were primarily focused on transcriptional changes affecting steady-state overall gene expression levels using microarray based approaches. A major missing component, however, has been the study of transcriptome changes at the post-transcriptional level, such as alternative splicing. Alternative splicing is a critical mechanism for expanding regulatory and functional diversity from a limited number of genes, and is particularly complex in the mammalian brain. Here we carried out a deep RNA-seq analysis of the BA4 (Brodmann area 4) motor cortex from seven human HD brains and seven controls to systematically discover aberrant alternative splicing events and characterize potential associated splicing factors in HD. We identified 593 differential alternative splicing events between HD and control brains. Using two expanded panels with a total of 108 BA4 tissues from patients and controls, we identified four splicing factors exhibiting significantly altered expression levels in HD patient brains. Moreover, follow-up molecular analyses of one splicing factor PTBP1 revealed its impact on disease-associated splicing patterns in HD. Collectively, our data provide genomic evidence for widespread splicing dysregulation in HD brains, and suggest the role of aberrant alternative splicing in the pathogenesis of HD.
亨廷顿舞蹈症(HD)是一种常染色体显性神经退行性疾病,由编码亨廷顿蛋白(HTT)的基因中的CAG重复扩增引起。转录组失调是HD发病机制的一个主要特征,大量针对人类HD患者和小鼠模型组织的基因表达谱研究揭示了这一点。这些研究主要集中于使用基于微阵列的方法来检测影响稳态整体基因表达水平的转录变化。然而,一个主要缺失的部分是对转录后水平的转录组变化的研究,比如可变剪接。可变剪接是一种从有限数量的基因扩展调控和功能多样性的关键机制,在哺乳动物大脑中尤为复杂。在这里,我们对来自7个HD患者大脑和7个对照的BA4(布罗德曼4区)运动皮层进行了深度RNA测序分析,以系统地发现异常可变剪接事件,并鉴定HD中潜在的相关剪接因子。我们在HD和对照大脑之间鉴定出593个差异可变剪接事件。使用两个扩展样本组,其中共有来自患者和对照的108个BA4组织,我们鉴定出4个在HD患者大脑中表达水平显著改变的剪接因子。此外,对一个剪接因子PTBP1的后续分子分析揭示了其对HD中疾病相关剪接模式的影响。总的来说,我们的数据为HD大脑中广泛存在的剪接失调提供了基因组证据,并提示异常可变剪接在HD发病机制中的作用。