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| Status |
Public on May 19, 2024 |
| Title |
Histone lactylation-mediated NSUN2 transcriptional activation dictates choroidal neovascularization through promoting AKAP2 mRNA expression [RNA-seq] |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
Other
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| 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.
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| Overall design |
RNA-seq data related to gene NSUN2 in human umbilical vein endothelial cell
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| Contributor(s) |
Zuo S, Li L, Lu L, Fan X |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
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| Submission date |
May 16, 2024 |
| Last update date |
May 19, 2024 |
| Contact name |
Sipeng Zuo |
| E-mail(s) |
Zsp2019@sjtu.edu.cn
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| Organization name |
Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine
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| Street address |
No. 12, Lane 833, Zhizaoju Road, Huangpu District
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| City |
Shanghai |
| State/province |
Shanghai |
| ZIP/Postal code |
200023 |
| Country |
China |
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| Platforms (2) |
| GPL21290 |
Illumina HiSeq 3000 (Homo sapiens) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
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| Samples (12)
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| GSM8272391 |
HUVEC, Hypoxia, RNA-seq [Treated-2] |
| GSM8272392 |
HUVEC, shCtrl, meRIP-seq [Control1.Input] |
| GSM8272393 |
HUVEC, shCtrl, meRIP-seq [Control2.Input] |
| GSM8272394 |
HUVEC, shCtrl, meRIP-seq [Control1.IP] |
| GSM8272395 |
HUVEC, shCtrl, meRIP-seq [Control2.IP] |
| GSM8272396 |
HUVEC, shNSUN2, meRIP-seq [Sh1.Input] |
| GSM8272397 |
HUVEC, shNSUN2, meRIP-seq [Sh2.Input] |
| GSM8272398 |
HUVEC, shNSUN2, meRIP-seq [Sh1.IP] |
| GSM8272399 |
HUVEC, shNSUN2, meRIP-seq [Sh2.IP] |
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| Relations |
| BioProject |
PRJNA1112368 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE267662_Differentially_Expressed_genes.xlsx |
114.5 Kb |
(ftp)(http) |
XLSX |
| GSE267662_Differentially_methylated_sites.mRNA.xlsx |
7.2 Mb |
(ftp)(http) |
XLSX |
| GSE267662_treated-vs-NC-all.gene.xls.gz |
4.5 Mb |
(ftp)(http) |
XLS |
SRA Run Selector |
| Raw data are available in SRA |
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