Related Experiment Video
Updated: May 22, 2025
Myeloid Innate Signaling Pathway Regulation by MALT1 Paracaspase Activity
Published on: January 7, 2019
Role of DNMT3a expression and nuclear translocation under ELAVL1 mediation for dendritic cell function and Th17/Treg
Background:
Chronic obstructive pulmonary disease (COPD) is a leading cause of morbidity and mortality worldwide. The DNA methyltransferase DNMT3a has been implicated in COPD, however its upstream regulation and downstream mechanisms remain unclear.
Methods:
Relative mRNA and protein levels of indicated genes in lung tissues and dendritic cells (DCs) were tested via qRT-PCR and western blot, respectively. Cellular distribution of DNMT3a in DCs was determined by immunofluorescence staining. COPD mouse model was established by exposing mice to cigarette smoke (CS) via nose. The Th17/Treg cell ratio was examined using flow cytometry. Production of indicated cytokines was assessed by corresponding commercial ELISA kit. Interplay between DACH1 and c-Jun was verified by Co-immunoprecipitation, ChIP and luciferase reporter assays. Methylation level of DACH1 was tested by methylation specific PCR.
Results:
DNMT3a expression was upregulated and negatively correlated with lung function in COPD patients. CS exposure increased pulmonary DNMT3a in mice. DNMT3a was predominantly expressed in the nucleus and CS exposure promoted its translocation to cytoplasm. RNA binding protein ELAVL1 upregulated DNMT3a expression, induced its nuclear translocation and increased its enzyme activity. DNMT3a promoted Th17 differentiation while inhibited Treg differentiation. DNMT3a methylated DACH1 and inhibited its expression, resulting in c-Jun pathway activation. In vivo DNMT3a knockdown ameliorated lung injury and Th17/Treg imbalance in COPD mice.
Conclusion:
This study suggests that ELAVL1 regulates DNMT3a expression and nuclear translocation to modulate dendritic cell function and Th17/Treg balance through DACH1/c-Jun pathway in COPD.
Insights
The RNA binding protein ELAVL1 upregulates DNMT3a, impacting dendritic cell function and T cell balance in chronic obstructive pulmonary disease (COPD). DNMT3a inhibition ameliorates lung injury in COPD mouse models.
Area of Science:
- Immunology
- Molecular Biology
- Pulmonology
Background:
- Chronic obstructive pulmonary disease (COPD) poses a significant global health burden.
- The role of DNA methyltransferase DNMT3a in COPD pathogenesis is recognized, but its regulatory mechanisms and downstream effects require elucidation.
Purpose of the Study:
- To investigate the upstream regulation and downstream mechanisms of DNMT3a in the context of COPD.
- To explore the impact of DNMT3a on dendritic cell function and T helper 17 (Th17)/regulatory T cell (Treg) balance.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) and western blotting to assess gene and protein expression in lung tissues and dendritic cells (DCs).
- Immunofluorescence staining for DNMT3a cellular distribution, flow cytometry for Th17/Treg cell ratios, and ELISA for cytokine production.
- Co-immunoprecipitation, ChIP, luciferase reporter assays, and methylation-specific PCR to investigate molecular interactions and DNA methylation.
Main Results:
- DNMT3a expression was elevated and inversely correlated with lung function in COPD patients and mice exposed to cigarette smoke (CS).
- ELAVL1 upregulated DNMT3a expression, promoted its nuclear translocation, and increased its enzymatic activity, leading to Th17 differentiation promotion and Treg differentiation inhibition.
- DNMT3a methylated and inhibited DACH1 expression, activating the c-Jun pathway. In vivo DNMT3a knockdown improved lung injury and Th17/Treg imbalance in COPD mice.
Conclusions:
- ELAVL1 regulates DNMT3a expression and localization, influencing dendritic cell function and Th17/Treg balance via the DACH1/c-Jun pathway in COPD.
- Targeting the ELAVL1/DNMT3a/DACH1/c-Jun axis presents a potential therapeutic strategy for COPD.
