Kumar Ashish, Zhang Jennifer, Tallaksen-Greene Sara, Crowley Michael R, Crossman David K, Morton A Jennifer, Van Groen Thomas, Kadish Inga, Albin Roger L, Lesort Mathieu, Detloff Peter J
Department of Cell, Developmental and Integrative Biology.
Department of Biochemistry and Molecular Genetics.
Hum Mol Genet. 2016 Apr 15;25(8):1619-36. doi: 10.1093/hmg/ddw040. Epub 2016 Feb 14.
Identifying molecular drivers of pathology provides potential therapeutic targets. Differentiating between drivers and coincidental molecular alterations presents a major challenge. Variation unrelated to pathology further complicates transcriptomic, proteomic and metabolomic studies which measure large numbers of individual molecules. To overcome these challenges towards the goal of determining drivers of Huntington's disease (HD), we generated an allelic series of HD knock-in mice with graded levels of phenotypic severity for comparison with molecular alterations. RNA-sequencing analysis of this series reveals high numbers of transcripts with level alterations that do not correlate with phenotypic severity. These discorrelated molecular changes are unlikely to be drivers of pathology allowing an exclusion-based strategy to provide a short list of driver candidates. Further analysis of the data shows that a majority of transcript level changes in HD knock-in mice involve alteration of the rate of mRNA processing and/or degradation rather than solely being due to alteration of transcription rate. The overall strategy described can be applied to assess the influence of any molecular change on pathology for diseases where different mutations cause graded phenotypic severity.
识别病理学的分子驱动因素可提供潜在的治疗靶点。区分驱动因素和偶然的分子改变是一项重大挑战。与病理学无关的变异使测量大量单个分子的转录组学、蛋白质组学和代谢组学研究更加复杂。为了克服这些挑战以实现确定亨廷顿舞蹈病（HD）驱动因素的目标，我们构建了一系列具有不同程度表型严重程度的HD基因敲入小鼠等位基因系列，以便与分子改变进行比较。对该系列小鼠进行RNA测序分析发现，大量转录本水平发生改变，但这些改变与表型严重程度无关。这些不相关的分子变化不太可能是病理学的驱动因素，因此可以采用基于排除法的策略来列出候选驱动因素的简短清单。对数据的进一步分析表明，HD基因敲入小鼠中大多数转录本水平的变化涉及mRNA加工和/或降解速率的改变，而不仅仅是由于转录速率的改变。所描述的总体策略可应用于评估任何分子变化对不同突变导致表型严重程度分级的疾病病理学的影响。
