Multi-dimensional Multi-omics Integrative Study to Identify Target Genes for Ischemic Stroke and Related Chronic Pain

Chronic pain is the most common complications and after-effects for ischemic stroke. Through exploring immune related cell death target genes of ischemic stroke is essential for understanding ischemic stroke and chronic pain complications. Referred to three types of immune related cell deaths' marker genes, mRNA and microRNA transcriptomics data from mice MCAO model were firstly analyzed through multi algorithms. Then, screening common gene with brain chronic pain related dataset. At the single-cell level, we performed immune cell identification and differentially expressional analysis for entire stroke brain environment and pseudo-time analysis for candidate immune cells. Based on GWAS and eQTLs, colocalization analysis, and drug target mendelian randomization methods were used to evaluate causal relationships and drug target effects. Furthermore, to explore spatial characters spatial transcriptomic analysis was conducted. At last, PCR experiments in animal model were conducted. Cell death state is positively correlated with immune infiltration degree. Five core mRNAs, S100a6, Anxa3, Ncf4, Capg, and Arpc1b, and key microRNA, miR-298-5p, were screened as biomarkers for immune related cell death. Among them, S100a6 play key roles. Toll like receptor pathway and CD4⁺ _γδ T cells were identified as core immune pathway and cells. By comparing with chronic pain GWAS results, S100a6 is screened as promising target. In single-cell analysis, S100a6 participated in CD4⁺ _γδ T cells differentiation and immune activation on IS. Drug target MR analysis showed that activation of S100a6 was able to decrease 23-54% probabilities to develop into IS. Furtherly, S100a6 gene could balance the negative effects of Cd4 expressed immune cells and protect neuronal function in brain injury spatial zone. In PCR experiment, differentially expressed level of five core genes got proved. In conclusion, our study revealed S100a6 played causal protective roles for ischemic stroke and related chronic pain, could be seen as potential drug target.