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Ciao3 knockout causes acute lung injury through immune activation using single cell sequencing
Jie Peng1, Chunying Wang1, Ting Liu1
1School of Clinical Medicine, the First Affiliated Hospital of Chengdu Medical College, Chengdu, Sichuan Province, 610500, PR China.
Background:
CIAO3 mutation is identified as the causative gene for diffuse pulmonary arteriovenous malformations (dPAVMs). A homozygote patient is follow-up and she is suffering from a cytokine storm. We conduct Ciao3 conditional knockout in endothelial cells (EC-CKO) of mice to clarify its potential roles.
Methods:
The cytokine levels of the homozygote patient and the EC-CKO mice are measured. Bulk RNA sequencing (RNA-seq) and single cell RNA sequencing (scRNA-seq) are performed. R Seurat package and cellmarker 2.0 are used to cluster and annotate the data. The differential, enrichment, protein interaction, and cell communication analyses are conducted.
Results:
The patient is suffering from a cytokine storm. Ciao3 knockout contributes to increased cytokine levels and acute lung injury (ALI) in mice, indicating activation of immune functions. Inhibition of TNF-α reverses the development of ALI, which reveals that the ALI is caused by inflammatory activation. RNA-seq of whole blood cell shows that inflammatory genes and pathways are upregulated in EC-CKO mice, and inhibition of TNF-α improves the effect of Ciao3 knockout. scRNA-seq of lung tissues displays complex changes in immune cell populations, which leads to abnormal cell-cell communication between ECs and immune cells. Outgoing and incoming signaling analyses suggest that CypA, FN1, and THBS signaling pathway are the main changed pathways, and the monocytes and macrophages are the central effector in EC-CKO mice. Upregulation of CypA and infiltration of macrophages are confirmed by immunohistochemistry.
Conclusion:
Ciao3 knockout causes ALI through immune activation in mice. The monocytes and macrophages are the central effector and should be further researched.
Insights
Ciao3 knockout in mice triggers a cytokine storm and acute lung injury (ALI) by activating immune cells, particularly monocytes and macrophages. Inhibiting TNF-α can reverse these effects, suggesting a therapeutic target for lung inflammation.
Area of Science:
- Vascular Biology
- Immunology
- Genetics
Background:
- The CIAO3 mutation is linked to diffuse pulmonary arteriovenous malformations (dPAVMs).
- A patient with a CIAO3 mutation experienced a cytokine storm.
- This study investigates the role of CIAO3 in endothelial cells (ECs).
Purpose of the Study:
- To clarify the potential roles of CIAO3 in endothelial cells.
- To investigate the link between CIAO3 mutation, cytokine storm, and acute lung injury (ALI).
- To identify the immune cells and pathways involved in CIAO3-related lung pathology.
Main Methods:
- Conditional knockout of Ciao3 in mouse endothelial cells (EC-CKO).
- Cytokine level measurement, bulk and single-cell RNA sequencing (RNA-seq, scRNA-seq).
- Bioinformatic analyses including differential gene expression, pathway enrichment, protein-protein interaction, and cell communication analysis.
Main Results:
- Ciao3 knockout in mice led to increased cytokine levels and acute lung injury (ALI), indicative of immune activation.
- Tumor necrosis factor-alpha (TNF-α) inhibition reversed ALI, confirming inflammatory activation as the cause.
- scRNA-seq revealed altered immune cell populations and abnormal EC-immune cell communication, with monocytes and macrophages identified as key effectors.
- Upregulation of CypA and macrophage infiltration were confirmed via immunohistochemistry.
Conclusions:
- Ciao3 knockout induces ALI in mice through immune activation.
- Monocytes and macrophages are central effectors in this process.
- Targeting these immune cells or related pathways may offer therapeutic strategies for CIAO3-associated lung diseases.
