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Identification and validation of neuroinflammation related lncRNA PVT1 with transcriptome-wide analysis in cerebral
Xiaochan Xiao1, Huodan Yu2, Haichuan Zhou3
1Department of Critical Care Medicine, Renmin Hospital of Wuhan University, Wuhan, China.
Background:
The cerebral ischemia/reperfusion injury (CIRI) is an essential pathological process of ischemic stroke (IS). Secondary neuroinflammation exacerbate neuronal damage following CIRI. To identify long non-coding RNAs (lncRNAs) implicated in neuroinflammation subsequent to CIRI would significantly advance the development of potential therapeutic interventions.
Methods:
Through comprehensive analysis of whole-genome RNA-seq profiles in focal ischemic mice models, we identified differentially expressed genes utilizing Gene Ontology term enrichment, Kyoto Encyclopedia of Genes and Genomes pathway analysis, and gene set enrichment analysis. We further implemented immune cell infiltration deconvolution, constructed protein-protein interaction networks, and performed co-expression network analysis for lncRNA screening. Subsequently, we established the mice model with lncRNA PVT1 knockdown prior to CIRI induction. Quantitative assessment of cytokine levels was conducted using enzyme-linked immunosorbent assay, while morphological alterations were evaluated through hematoxylin-eosin staining. And T cell infiltration in cerebral tissues was detected with immunofluorescence analysis.
Results:
Enrichment analysis demonstrated that differentially expressed mRNAs were implicated in neuroinflammation following cerebral ischemic. Through immune deconvolution analysis, we observed a increased levels in the CD4 + and CD8 + T cells proportion of cerebral ischemic groups compared with control groups. It identified five hub lncRNAs (AI662270, AU020206, Gm20667, PVT1 and Mir142hg) exhibiting significant correlations with the expression of proinflammatory factors. Notably, PVT1 demonstrated the strongest correlation coefficient with pro-inflammatory factor mRNA expression levels. The vivo experimental validation revealed aberrantly elevated PVT1 expression following CIRI. Importantly, PVT1 knockdown substantially ameliorated CIRI through the reduction of activated T cell infiltration and pro-inflammatory cytokine secretion.
Conclusion:
The identified lncRNA PVT1 correlated with the activated T cell infiltration and pro-inflammatory cytokine secretion, which could be treatment target for neuroinflammation in CIRI.
