Single-cell transcriptomics of mammalian prion diseases identifies dynamic gene signatures shared between species [mouse_scRNAseq]. Single-cell transcriptomics of mammalian prion diseases identifies dynamic gene signatures shared between species [mouse_scRNAseq]

Mammalian prion diseases are fatal and transmissible neurological conditions caused by the propagation of prions, self-replicating multimeric assemblies of misfolded forms of host cellular prion protein. Despite extensive studies investigating the changes in transcriptional profiles in prion diseases the mechanisms by which prion diseases induce cellular toxicity, including changes in gene expression profiles are yet to be fully characterized. Here, we took advantage of the recent developments in single-cell technologies and performed an unbiased whole-transcriptome single-nucleus transcriptomic analysis in prion disease. Overall design: FVB inbred mice were intracerebrally inoculated with infectious RML brain homogenate, uninfected CD1 brain homogenate, or PBS and culled at 5 time points (20 dpi, 40 dpi , 80 dpi, 120 dpi, and when scrapie sickness was confirmed). Half brains were flash frozen and stored at -80oC until further processing. For each sample, the brain was left to thaw and a section of the frontal lobe was cut and processed for scRNA-seq using the SPLiT-seq protocol.

哺乳动物朊病毒病（prion diseases）是一类致死性、可传播的神经系统疾病，由朊病毒（prions）的增殖所引发：朊病毒即宿主细胞朊蛋白（host cellular prion protein）错误折叠后形成的自我复制多聚体组装体。尽管已有诸多研究针对朊病毒病的转录组特征变化展开探索，但该病诱导细胞毒性的具体机制（包括基因表达谱改变在内）仍未得到完全阐明。本研究借助近期单细胞技术的发展成果，对朊病毒病开展了无偏倚的全转录组单细胞核转录组分析。实验设计：将FVB近交小鼠经颅内分别接种传染性RML脑匀浆、未感染的CD1小鼠脑匀浆或磷酸盐缓冲液（PBS），并于5个时间点处死采样：接种后20天（20 dpi）、40天（40 dpi）、80天（80 dpi）、120天（120 dpi），以及确诊羊瘙痒症发病时。将获取的半数脑组织快速冷冻并保存于-80℃，直至后续处理。针对每一份样本，将脑组织解冻后切取额叶组织，采用SPLiT-seq技术进行单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）。