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Published on: March 21, 2021
Mito-TEMPO attenuates LPS-induced acute lung injury accompanied by ferroptosis suppression and reduced HSP90-STING
Yun Yao1, Wei Guo1, Yujia Tian2
1Department of Respiratory and Critical Care Medicine, Lujiang County People's Hospital, Hefei 231500, China.
Background:
Acute lung injury (ALI) lacks effective pharmacological treatments. Ferroptosis and stimulator of interferon genes (STING)-dependent immune activation contribute to ALI, but whether the heat shock protein 90 (HSP90)-STING interface can be modulated to limit ferroptotic injury remains unclear. This study investigated whether Mito-TEMPO protects against lipopolysaccharide (LPS)-induced ALI in relation to the HSP90-STING-ferroptosis axis.
Methods:
LPS-induced ALI was established in wild-type and STING-deficient (STING-/-) mice with intratracheal Mito-TEMPO pretreatment. Lung injury, vascular permeability, inflammation, and ferroptosis were evaluated by histology, bronchoalveolar lavage fluid (BALF) analysis, ELISA, qRT-PCR, Western blotting, immunohistochemistry, and immunofluorescence. LPS-stimulated BEAS-2B cells were used for in vitro validation, and additional A549 cells with STING overexpression were employed to further validate STING-dependent signaling and ferroptosis-related mechanisms. Network pharmacology, molecular docking, co-immunoprecipitation, and colocalization analyses were performed to examine HSP90-STING signaling.
Results:
Mito-TEMPO alleviated LPS-induced pulmonary edema, inflammatory infiltration, tissue injury, BALF protein accumulation, BALF cellularity, and IL-6, IL-1β, and TNF-α expression. It also reversed ferroptosis-associated changes by restoring GPX4 and FTH1 while reducing NCOA4, PTGS2, and 4-HNE, with decreased lipid peroxidation in SFTPC+ alveolar type II (AT2) cells. Similar anti-inflammatory and anti-ferroptotic effects were observed in BEAS-2B cells. Network pharmacology and docking analyses suggested HSP90 as a potential Mito-TEMPO-associated target. Co-immunoprecipitation and immunofluorescence analyses showed reduced HSP90-STING interaction after Mito-TEMPO treatment, accompanied by decreased levels of phosphorylated TBK1 and IRF3. In STING-deficient mice, LPS-induced injury was attenuated and the additional protective effects of Mito-TEMPO were diminished. In A549 cells, STING overexpression aggravated LPS-induced inflammatory and ferroptosis-related changes and partially attenuated the protective effects of Mito-TEMPO.
Conclusions:
Mito-TEMPO attenuates LPS-induced experimental ALI in association with reduced inflammation, ferroptosis-related molecular alterations, reduced HSP90-STING interaction, and attenuated downstream STING-TBK1-IRF3 signaling. These findings suggest that the HSP90-STING-ferroptosis axis may represent a mechanistically relevant pathway warranting further investigation.
Insights
Mito-TEMPO treatment reduces inflammation and ferroptosis in acute lung injury (ALI) by targeting the heat shock protein 90 (HSP90)-stimulator of interferon genes (STING) pathway. This suggests a novel therapeutic strategy for ALI.
Area of Science:
- Investigates the intersection of immunology, cell death pathways, and pharmacology in the context of lung injury.
- Focuses on the role of ferroptosis and STING-dependent immune responses in acute lung injury (ALI).
- Explores the potential of targeting the HSP90-STING-ferroptosis axis for therapeutic intervention.
Background:
- Acute lung injury (ALI) currently lacks effective pharmacological treatments.
- Ferroptosis and stimulator of interferon genes (STING)-dependent immune activation are implicated in ALI pathogenesis.
- The potential to modulate the heat shock protein 90 (HSP90)-STING interface to mitigate ferroptotic injury in ALI remains unexplored.
Purpose of the Study:
- To investigate the protective effects of Mito-TEMPO against lipopolysaccharide (LPS)-induced ALI.
- To elucidate the role of the HSP90-STING-ferroptosis axis in mediating these protective effects.
- To evaluate whether Mito-TEMPO can modulate the HSP90-STING interaction and downstream signaling pathways.
Main Methods:
- Established LPS-induced ALI in wild-type and STING-deficient mice, with intratracheal Mito-TEMPO pretreatment.
- Assessed lung injury, vascular permeability, inflammation, and ferroptosis using histological, biochemical, and molecular analyses.
- Utilized in vitro cell models (BEAS-2B, A549) and computational methods (network pharmacology, molecular docking) to validate findings and explore mechanisms.
Main Results:
- Mito-TEMPO significantly alleviated LPS-induced lung injury, edema, inflammation (reduced IL-6, IL-1β, TNF-α), and ferroptosis markers.
- Mito-TEMPO treatment reduced HSP90-STING interaction and downstream STING-TBK1-IRF3 signaling.
- Protective effects were diminished in STING-deficient mice, and STING overexpression exacerbated injury, confirming STING-dependent mechanisms.
Conclusions:
- Mito-TEMPO demonstrates significant protective effects against experimental ALI.
- These effects are associated with reduced inflammation, ferroptosis, and modulation of the HSP90-STING-TBK1-IRF3 signaling axis.
- The HSP90-STING-ferroptosis pathway represents a promising therapeutic target for ALI.
