1 Department of Pediatrics, Biological Sciences Division, The University of Chicago, IL, USA.
2 Center for Research Informatics, Biological Sciences Division, The University of Chicago, IL, USA.
ASN Neuro. 2019 Jan-Dec;11:1759091419843393. doi: 10.1177/1759091419843393.
In humans, homozygous mutations in the TPP1 gene results in loss of tripeptidyl peptidase 1 (TPP1) enzymatic activity, leading to late infantile neuronal ceroid lipofuscinoses disease. Using a mouse model that targets the Tpp1 gene and recapitulates the pathology and clinical features of the human disease, we analyzed end-stage (4 months) transcriptional changes associated with lack of TPP1 activity. Using RNA sequencing technology, Tpp1 expression changes in the forebrain/midbrain and cerebellum of 4-month-old homozygotes were compared with strain-related controls. Transcriptional changes were found in 510 and 1,550 gene transcripts in forebrain/midbrain and cerebellum, respectively, from Tpp1-deficient brain tissues when compared with age-matched controls. Analysis of the differentially expressed genes using the Ingenuity™ pathway software, revealed increased neuroinflammation activity in microglia and astrocytes that could lead to neuronal dysfunction, particularly in the cerebellum. We also observed upregulation in the production of nitric oxide and reactive oxygen species; activation of leukocyte extravasation signals and complement pathways; and downregulation of major transcription factors involved in control of circadian rhythm. Several of these expression changes were confirmed by independent quantitative polymerase chain reaction and histological analysis by mRNA in situ hybridization, which allowed for an in-depth anatomical analysis of the pathology and provided independent confirmation of at least two of the major networks affected in this model. The identification of differentially expressed genes has revealed new lines of investigation for this complex disorder that may lead to novel therapeutic targets.
在人类中,TPP1 基因的纯合突变导致三肽基肽酶 1(TPP1)酶活性丧失,导致晚发性婴儿神经元蜡样脂褐质沉积症。使用靶向 Tpp1 基因并重现人类疾病的病理学和临床特征的小鼠模型,我们分析了与缺乏 TPP1 活性相关的终末期(4 个月)转录变化。使用 RNA 测序技术,将 4 个月大的纯合子的前脑/中脑和小脑中的 Tpp1 表达变化与与菌株相关的对照进行了比较。与年龄匹配的对照相比,在 Tpp1 缺陷脑组织中分别在 510 和 1550 个基因转录本中发现了转录变化。使用 Ingenuity™途径软件对差异表达基因进行分析,发现小胶质细胞和星形胶质细胞中的神经炎症活性增加,这可能导致神经元功能障碍,特别是在小脑。我们还观察到一氧化氮和活性氧的产生增加;白细胞渗出信号和补体途径的激活;以及涉及昼夜节律控制的主要转录因子的下调。这些表达变化中的几个通过独立的定量聚合酶链反应和 mRNA 原位杂交的组织学分析得到了证实,这允许对病理学进行深入的解剖学分析,并为该模型中受影响的至少两个主要网络提供了独立的确认。差异表达基因的鉴定为这种复杂疾病的研究提供了新的线索,可能为新的治疗靶点提供依据。