Exosome-related genes influence the progression of stroke through neuroinflammatory responses

Boyan Zhao^1,2Wu Jianing¹Han Mingyang¹Xuan Rong¹Jin Jin²Shiya Liu²Cheng Zhang³Ruotian Zhang⁴Xin Chen⁴Fei Peng⁵Xingli Dong^6*Shiguang Zhao^1,2*

• ¹Department of Neurosurgery, Shenzhen University General Hospital, Shenzhen, China
• ²School of Medicine, Shenzhen University, Shenzhen, Guangdong Province, China
• ³University of Toronto Scarborough, Toronto, Ontario, Canada
• ⁴Department of Neurosurgery, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang Province, China
• ⁵Department of Neurosurgery, Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong Province, China
• ⁶Shenzhen University General Hospital, Shenzhen, China

Ischemic stroke (IS) ranks among the top causes of mortality and disability globally. Exosomes exert a crucial effect on maintaining a complex regulatory balance with neuroinflammation in IS. Hence, this research aimed to elucidate the roles of exosome-related genes IS. We integrated data from five IS-related datasets from the Gene Expression Omnibus (GEO) database and exosome-related genes from ExoCarta. The least absolute shrinkage and selection operator regression and random forest models were performed to detect feature genes. Search Tool for the Retrieval of Interacting Genes and Cytoscape were employed to recognize the hub genes. Enrichment analyses were conducted to examine biological processes. CIBERSORT and MCPcounter were applied to assess immune infiltration, and Principal Component Analysis was utilized to explore the associations of feature genes and hub genes with immune cells. After identified different cell types, we analyzed differentiation, developmental trajectory, and interactions of the cell populations. Middle cerebral artery occlusion models were conducted on mice, followed by quantitative polymerase chain reaction to assess the expression levels of each hub gene. We identified 13 feature genes and 10 hub genes. Through qPCR, LGALS3, CD36, TLR2, ICAM1, and CD14 were significantly upregulated after Middle Cerebral Artery Occlusion surgery. Hub genes were significantly involved in inflammatory responses, as well as chemokine signaling and JAK-STAT signaling. Immuno-infiltration analysis revealed significant differences in immune cell populations between IS and controls. Additionally, neutrophils and monocytes/macrophages were positively correlated with CD14 and LGALS3, respectively. Single-cell analysis revealed 19 cell subpopulations with detailed pseudo-time trajectory predictions, highlighting the developmental importance of MG2 microglial cells. In conclusion, our results illuminate exosomal genes, including LGALS3 and CD14, participate in the progression of IS through neuroinflammation, as well as highlight potential therapeutics to mitigate IS injury.In this research, we sought to uncover the molecular mechanisms contributing to IS by integrating multiomics data with advanced analytical methods. We concentrated on the involvement of exosomerelated genes in IS and highlighted the importance of neuroinflammation.Materials and Methods Overall analysis flow chart.