@article{XIONG2026, 
author = {SHAOFENG XIONG and WENKAI LV and GUOSHENG CAO and LONGSHENG FU and WEN LIU and MENGFAN LEI and YANNI LV},
title = {Integrative Analysis of scRNA-Seq and Bulk RNA-Seq Reveals Novel Transcription Factor Regulating Endothelial Heterogeneity Induced by Lrg1 Following Cerebral Ischemia-Reperfusion},
year = {2026},
journal = {BIOCELL},
volume = {50},
number = {1},
pages = {12},
keywords = {Single-cell RNA sequencing, endothelial heterogeneity, leucine-rich alpha-2 glycoprotein 1, zinc finger MIZ-type containing 1 Protein, Frizzled 3, transcription factor, cerebral ischemia reperfusion},
url = {https://www.sciopen.com/article/10.32604/biocell.2025.073401},
doi = {10.32604/biocell.2025.073401},
abstract = {ObjectiveLeucine-rich alpha-2 glycoprotein 1 (Lrg1) could regulate diverse cells in cerebral ischemia-reperfusion. Our study seeks to uncover Lrg1’s impact on endothelial cell heterogeneity via differentiation pathways and transcription factors.MethodThe CSOmap model measured cell-to-brain-center distances using single-cell RNA sequencing (scRNA-seq) data in middle cerebral artery occlusion reperfusion (MCAO/R). Monocle2 mapped endothelial differentiation paths. Gene set enrichment analysis (GSEA) analyzed endothelial subcluster variations. Database searches revealed a zinc finger MIZ-type containing 1 protein-frizzled 3 (Zmiz1-Fzd3) promoter interaction. Endothelial cells were transfected with a Fzd3 promoter-luciferase plasmid. Polymerase chain reaction (PCR) and western blotting assessed MCAO/R or Zmiz1 overexpression effects on Fzd3-related mRNA and proteins. A retroviral vector carrying Zmiz1 was injected into the brains of mice to study its effect on Fzd3.ResultLrg1−/− mice exhibited elevated cell adhesion proteins and decreased microvascular leakage after MCAO/R. CSOmap showed widened astrocyte spacing in these mice. RSS revealed Zmiz1 overexpression in MCAO/R+Lrg1−/− mice. MCAO/R and pcDNA3-Zmiz1 transfection both enhanced luciferase activity with Fzd3, indicating Zmiz1 binding to Fzd3. Retroviral Zmiz1 injection or knockdown disrupted ischemic brain tight junctions, highlighting Zmiz1’s key role in blood-brain barrier protection, likely through Fzd3 pathway modulation.ConclusionThe findings indicate Lrg1 knockout induces endothelial differentiation by activating Zmiz1, which is crucial for maintaining blood-brain barrier function, possibly via modulating the Fzd3 pathway.}
}