Combination of single-nucleus and bulk RNA-seq reveals the molecular mechanism of thalamus haemorrhage-induced central poststroke pain.
Combination of single-nucleus and bulk RNA-seq reveals the molecular mechanism of thalamus haemorrhage-induced central poststroke pain.
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DOI：
10.3389/fimmu.2023.1174008
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发表时间：
2023
影响因子：
7.3
通讯作者：
中科院分区：
文献类型：
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作者：
丘脑出血（TH）引起的中枢性卒中后疼痛（CPSP）可为持续性或间歇性，并伴有感觉异常，严重影响患者的生活质量。深入了解CPSP机制和治疗策略需要更深入地了解丘脑的分子过程。在这里，使用单核RNA测序（snRNA-seq），我们对32332个脑细胞的转录组进行了测序，这揭示了小鼠四个丘脑样本中总共有四种主要的细胞类型。与对照组相比，实验组对机械、热和冷刺激的敏感性更高，小胶质细胞数量增加，神经元数量减少。我们分析了从批量RNA测序（批量RNA-seq）数据中获得的差异表达基因和神经元标记基因的集合，发现Apoe，Abca 1和Hexb是通过免疫荧光（IF）验证的关键基因。免疫浸润分析发现这些关键基因与巨噬细胞、T细胞、相关趋化因子、免疫刺激因子和受体密切相关。基因本体论（GO）富集分析也表明，关键基因富集的生物过程，如蛋白质从核输出和蛋白质类小泛素化。总之，使用大规模snRNA-seq，我们已经确定了TH后大脑中的转录和细胞多样性。我们对丘脑内离散细胞类型和差异表达基因的鉴定可以促进新的CPSP疗法的开发。
Central poststroke pain (CPSP) induced by thalamic haemorrhage (TH) can be continuous or intermittent and is accompanied by paresthesia, which seriously affects patient quality of life. Advanced insights into CPSP mechanisms and therapeutic strategies require a deeper understanding of the molecular processes of the thalamus. Here, using single-nucleus RNA sequencing (snRNA-seq), we sequenced the transcriptomes of 32332 brain cells, which revealed a total of four major cell types within the four thalamic samples from mice. Compared with the control group, the experimental group possessed the higher sensitivity to mechanical, thermal, and cold stimuli, and increased microglia numbers and decreased neuron numbers. We analysed a collection of differentially expressed genes and neuronal marker genes obtained from bulk RNA sequencing (bulk RNA-seq) data and found that Apoe, Abca1, and Hexb were key genes verified by immunofluorescence (IF). Immune infiltration analysis found that these key genes were closely related to macrophages, T cells, related chemokines, immune stimulators and receptors. Gene Ontology (GO) enrichment analysis also showed that the key genes were enriched in biological processes such as protein export from nucleus and protein sumoylation. In summary, using large-scale snRNA-seq, we have defined the transcriptional and cellular diversity in the brain after TH. Our identification of discrete cell types and differentially expressed genes within the thalamus can facilitate the development of new CPSP therapeutics.
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DOI：
10.1007/978-1-0716-1162-3_20
发表时间：
2021-01-01
期刊：
DETECTION OF CELL DEATH MECHANISMS
影响因子：
--
作者：
Guy, Joe L.;Mor, Gil G.
通讯作者：
Mor, Gil G.
影响因子：
48
作者：
Habib N;Avraham-Davidi I;Basu A;Burks T;Shekhar K;Hofree M;Choudhury SR;Aguet F;Gelfand E;Ardlie K;Weitz DA;Rozenblatt-Rosen O;Zhang F;Regev A
通讯作者：
Regev A
影响因子：
6
作者：
Choi, Hanwool Ryan;Aktas, Adem;Bottros, Michael M.
通讯作者：
Bottros, Michael M.
神经炎症驱动的慢性疼痛的新兴目标。
影响因子：
120.1
作者：
Ji, Ru-Rong;Xu, Zhen-Zhong;Gao, Yong-Jing
通讯作者：
Gao, Yong-Jing
影响因子：
3.7
作者：
Bakken, Trygve E.;Hodge, Rebecca D.;Tasic, Bosiljka
通讯作者：
Tasic, Bosiljka