Comparative transcriptional profiling of motor neuron disorder-associated genes in various human cell culture models

Disease modelling requires appropriate cellular models that best mimic the underlying pathophysiology. Human origin and an adequate expression of the disease protein are pre-requisites that support information from a model to be meaningful. In this study we investigated expression profiles of (i) PBMCs and (ii) fibroblasts as patient derived cells as well as (iii) lymphoblasts and (iv) induced pluripotent stem cells (iPSC) as immortalized sources and (v) iPSC-derived cortical neurons to assess its aptitude to model motor neuron diseases (MNDs) including hereditary spastic paraplegia (HSP), amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA). We generated all five cell lines from two healthy donors and performed RNA sequencing to display expression patterns in MND-related genes. For the ten most common HSP genotypes we proved gene expression by qPCR. Depending on the specific MND gene we found largely different expression patterns. Out of 168 MND-related genes, 50 had their highest expression in iPSC-derived cortical neurons, 41 were most strongly expressed in fibroblasts, 26 in lymphoblasts, 22 in iPSCs and 14 in PBMCs. 15 MND-related genes were not detectable in any of the analyzed cell types. This study provides comprehensive information on expression of genes associated with a large spectrum of MNDs. Expression profiles can be used to inform on appropriate cell models for genotype specific motor neuron research. In total, 10 samples are analyzed, distributed over five different cell types with duplicates for each type.

疾病建模需要能够最优模拟潜在病理生理过程的适配细胞模型。人类细胞起源与疾病蛋白的足量表达，是确保模型输出信息具备科学意义的核心先决条件。本研究针对五类细胞的表达谱展开探究：(i) 外周血单个核细胞（PBMCs）与(ii) 成纤维细胞，均为患者来源细胞；(iii) 淋巴母细胞与(iv) 诱导多能干细胞（iPSC），属于永生化细胞来源；以及(v) 诱导多能干细胞分化得到的皮质神经元，以此评估各类细胞用于建模运动神经元病（MNDs）的适配性，所覆盖的疾病类型包括遗传性痉挛性截瘫（HSP）、肌萎缩侧索硬化（ALS）与脊髓性肌萎缩症（SMA）。我们从两名健康供体中成功构建上述全部五种细胞系，并通过RNA测序展示了运动神经元病相关基因的表达模式。针对十种最常见的遗传性痉挛性截瘫基因型，我们通过定量聚合酶链反应（qPCR）验证了其基因表达情况。根据特定运动神经元病基因的差异，我们观测到表达模式存在显著分歧。在168个运动神经元病相关基因中，50个在诱导多能干细胞分化的皮质神经元中表达量最高，41个在成纤维细胞中表达最强，26个在淋巴母细胞中，22个在诱导多能干细胞中，14个在外周血单个核细胞中。另有15个运动神经元病相关基因在所有分析的细胞类型中均未检出。本研究为涵盖大范围运动神经元病相关基因的表达特征提供了全面的数据支撑。其表达谱可用于指导针对特定基因型的运动神经元病研究，选择适配的细胞模型。本次研究共分析10份样本，分布于五种不同细胞类型中，每种细胞类型设置两份生物学重复样本。