Data_Sheet_4_Analysis of Age-Dependent Transcriptomic Changes in Response to Intracerebral Hemorrhage in Mice.xlsx

Age is a well-known risk factor that is independently associated with poor outcomes after intracerebral hemorrhage (ICH). However, the interrelationship between age and poor outcomes after ICH is not well defined. In this study, we aimed to investigate this relationship based on collagenase-induced ICH mice models. After being assessed neurological deficit 24 h after ICH, mice were euthanized and brain perihematomal tissues were used for RNA-sequencing (RNA-seq). And then the functions of differentially expressed genes (DEGs) identified by RNA-seq were analyzed using Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, Ingenuity Pathway Analysis (IPA) and protein-protein interaction (PPI) analysis. In addition, we performed real-time quantitative polymerase chain reaction (RT-qPCR) for validation of candidate DEGs. In the behavioral tests, aged mice presented significantly worse neurological function than young mice and greater weight loss than aged sham controls 24 h after ICH. In DEGs analysis, ICH affected the expression of more genes in young mice (2,337 DEGs) compared with aged mice (2,005 DEGs). We found aged mice exhibited increased brain inflammatory responses compared with young animals and ICH induced significant activation of the interferon-β (IFN-β) and IFN signaling pathways exclusively in aged mice. Moreover, further analysis demonstrated that ICH resulted in the activation of cytosolic DNA-sensing pathway with the production of downstream molecule type I IFN, and the response to type I IFN was more significant in aged mice than in young mice. In agreement with the results of RNA-seq, RT-qPCR indicated that the expression of candidate genes of cyclic GMP-AMP synthase (cGAS), Z-DNA-binding protein 1 (ZBP1), and IFN-β was significantly altered in aged mice after ICH. Taken together, our study indicated that compared to young animals, aged mice exhibit increased vulnerability to ICH and that the differences in transcriptional response patterns to ICH between young and aged mice. We believe that these findings will facilitate our understanding of ICH pathology and help to translate the results of preclinical studies into a clinical setting.

年龄是众所周知的独立风险因素，与脑出血（ICH）后的不良预后密切相关。然而，年龄与脑出血后不良预后的相互关系尚不明确。在本研究中，我们旨在基于胶原蛋白酶诱导的脑出血小鼠模型，探讨这一关系。在评估脑出血后24小时的神经功能缺损后，对小鼠进行安乐死，并使用脑出血周围的脑组织进行RNA测序（RNA-seq）。随后，通过基因本体（GO）分析、京都基因与基因组百科全书（KEGG）分析、灵感通路分析（IPA）和蛋白质-蛋白质相互作用（PPI）分析，对由RNA-seq鉴定的差异表达基因（DEGs）的功能进行分析。此外，我们还对候选DEGs进行了实时定量聚合酶链反应（RT-qPCR）以验证其表达。在行为测试中，老年小鼠在脑出血后24小时表现出比年轻小鼠更差的神经功能，以及比老年假手术对照组更大的体重减轻。在DEGs分析中，与老年小鼠（2,005个DEGs）相比，脑出血影响了年轻小鼠中更多基因的表达（2,337个DEGs）。我们发现，与年轻动物相比，老年小鼠表现出更高的脑炎症反应，且脑出血仅在老年小鼠中诱导了干扰素-β（IFN-β）和IFN信号通路的显著激活。此外，进一步分析表明，脑出血导致细胞质DNA感应途径的激活，产生下游分子I型IFN，且对I型IFN的反应在老年小鼠中比在年轻小鼠中更为显著。与RNA-seq的结果一致，RT-qPCR表明，在脑出血后，cGAS（环状GMP-AMP合酶）、ZBP1（Z-DNA结合蛋白1）和IFN-β候选基因的表达在老年小鼠中发生了显著改变。综上所述，我们的研究指出，与年轻动物相比，老年小鼠对脑出血的易感性更高，并且年轻和老年小鼠对脑出血的转录反应模式存在差异。我们相信，这些发现将有助于我们理解脑出血的病理学，并有助于将临床前研究结果转化为临床应用。