Soreq Lilach, Salomonis Nathan, Guffanti Alessandro, Bergman Hagai, Israel Zvi, Soreq Hermona
Department of Medical Neurobiology, The Hebrew University - Hadassah Medical School, Jerusalem 91120, Israel.
Department of Pediatrics, Division of Biomedical Informatics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA.
Genom Data. 2014 Nov 22;3:57-60. doi: 10.1016/j.gdata.2014.11.009. eCollection 2015 Mar.
Recent evidence demonstrates the power of RNA sequencing (RNA-Seq) for identifying valuable and urgently needed blood biomarkers and advancing both early and accurate detection of neurological diseases, and in particular Parkinson's disease (PD). RNA sequencing technology enables non-biased, high throughput, probe-independent inspection of expression data and high coverage and both quantification of global transcript levels as well as the detection of expressed exons and junctions given a sufficient sequencing depth (coverage). However, the analysis of sequencing data frequently presents a bottleneck. Tools for quantification of alternative splicing from sequenced libraries hardly exist at the present time, and methods that support multiple sequencing platforms are especially lacking. Here, we describe in details a whole RNA-Seq transcriptome dataset produced from PD patient's blood leukocytes. The samples were taken prior to, and following deep brain stimulation (DBS) treatment while being on stimulation and following 1 h of complete electrical stimulation cessation and from healthy control volunteers. We describe in detail the methodology applied for analyzing the RNA-Seq data including differential expression of long noncoding RNAs (lncRNAs). We also provide details of the corresponding analysis of in-depth splice isoform data from junction and exon reads, with the use of the software AltAnalyze. Both the RNA-Seq raw (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE42608) and analyzed data (https://www.synapse.org/#!Synapse:syn2805267) may be found valuable towards detection of novel blood biomarkers for PD.
最近的证据表明,RNA测序(RNA-Seq)在识别有价值且急需的血液生物标志物以及推动神经疾病尤其是帕金森病(PD)的早期准确检测方面具有强大作用。RNA测序技术能够对表达数据进行无偏倚、高通量、无需探针的检测,在足够的测序深度(覆盖度)下,可实现对全局转录水平的高覆盖度定量以及对表达外显子和连接点的检测。然而,测序数据的分析常常成为瓶颈。目前几乎不存在用于从测序文库中定量可变剪接的工具,尤其缺乏支持多种测序平台的方法。在此,我们详细描述了一个由PD患者血液白细胞产生的全RNA-Seq转录组数据集。样本采集于深部脑刺激(DBS)治疗前、治疗期间、治疗后1小时完全停止电刺激时,以及来自健康对照志愿者。我们详细描述了用于分析RNA-Seq数据的方法,包括长链非编码RNA(lncRNA)的差异表达。我们还提供了使用软件AltAnalyze对来自连接点和外显子 reads 的深度剪接异构体数据进行相应分析的详细信息。RNA-Seq原始数据(http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE42608)和分析后的数据(https://www.synapse.org/#!Synapse:syn2805267)对于检测PD的新型血液生物标志物可能具有价值。