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Series GSE267661 Query DataSets for GSE267661
Status Public on May 19, 2024
Title Histone lactylation-mediated NSUN2 transcriptional activation dictates choroidal neovascularization through promoting AKAP2 mRNA expression [CUT&Tag]
Organism Homo sapiens
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary Background: As a widespread post-transcriptional RNA modification, N5-methylcytosine (m5C) is implicated in a variety of cellular responses and processes that regulate RNA metabolism. Despite this, a clear understanding of m5C modification’s role and mechanism in angiogenesis is still lacking.
Methods: Single-cell RNA sequencing data was analyzed to determine expression of m5C methylase NSUN2. m5C levels were determined by mRNA isolation and anti-m5C dot blot in both hypoxia-induced endothelial cells (ECs) and laser-induced choroidal neovascularization (CNV). In addition, endothelial cell and endothelium‐specific NSUN2‐knockout mouse model were used to investigate the effect of NSUN2 silence on angiogenic phenotype. Genome-wide multiomics analyses were performed to identify the functional target of NSUN2, including proteomic analysis, transcriptome screening and m5C-methylated RNA immunoprecipitation sequencing (m5C-meRIP-seq). CUT&Tag sequencing was performed to test the histone lactylation signal in the promoter region of NSUN2. Finally, AAV-mediated short hairpin RNAi knockdown of NSUN2 gene expression (AAV-shNSUN2) was constructed to investigate the effect of inhibiting CNV.
Results: First, we discovered that m5C methylase NSUN2 expression level and mRNA m5C level were significantly higher in CNV-ECs than in normal ECs. NSUN2 knockdown in ECs inhibited proliferative, migration, and tube formation activities of ECs. Moreover, compared with EC NSUN2flox/flox mice, EC-specific NSUN2-deficient (EC NSUN2-/-) mice displayed less retinal vascular leakage after laser induction. Through multiomics analyses, we subsequently identified A-kinase anchoring protein 2 (AKAP2), a scaffolding protein which isolate Protein kinase A (PKA) to specific subcellular locations through binding to its regulatory subunit, as a downstream candidate target of NSUN2 in ECs. Overexpression of exogenous AKAP2 markedly reversed the inhibitory phenotypes in NSUN2-deficient ECs. Interestingly, laser induced NSUN2 up-regulation was driven by lactate-mediated lactylation on histone H3K18. In CNV models, AAV-mediated repression of NSUN2 modulated highly retinal vascular leakage and choroidal thickness.
Conclusion: Overall, our findings indicate that NSUN2 is a novel therapeutic target for choroidal neovascularization.
 
Overall design CUT&Tag-seq data related to gene NSUN2 in human umbilical vein endothelial cell
 
Contributor(s) Zuo S, Li L, Lu L, Fan X
Citation missing Has this study been published? Please login to update or notify GEO.
Submission date May 16, 2024
Last update date May 19, 2024
Contact name Sipeng Zuo
E-mail(s) Zsp2019@sjtu.edu.cn
Organization name Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine
Street address No. 12, Lane 833, Zhizaoju Road, Huangpu District
City Shanghai
State/province Shanghai
ZIP/Postal code 200023
Country China
 
Platforms (1)
GPL24676 Illumina NovaSeq 6000 (Homo sapiens)
Samples (4)
GSM8272384 HUVEC, ctrl [C_1]
GSM8272385 HUVEC, ctrl [C_1-igg]
GSM8272386 HUVEC, lactate [L_1]
Relations
BioProject PRJNA1112365

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Supplementary file Size Download File type/resource
GSE267661_All.xls.gz 217 b (ftp)(http) XLS
SRA Run SelectorHelp
Raw data are available in SRA

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