Chemoprevention of Lung Cancer with a Combination of Mitochondria-Targeted Compounds

Figure 1. Efficacy of Mito-HNK, Mito-LND, and their combination in the B[a]P-induced lung cancer model. (A) Structures of HNK, Mito-HNK, LND, and Mito-LND. (B) Efficacy on tumor load. (C) Body weights. (D–F) Plasma levels of liver enzymes and glucose (n = 5). Data are presented as mean ± SEM; *** p < 0.001.

Figure 2. Clustering analysis of scRNA-seq data from CD45− flow-sorted lung cells from control and Mito-HNK groups. (A) Lung adenoma cells were identified using scCancer software. Marker gene expression for (B) epithelial cells, (C) endothelial cells, and (D) fibroblasts. (E) Annotation of CD45− cells into the four identified cell populations.

Figure 3. Pathway changes between control and Mito-HNK groups in the CD45− cell populations. KEGG metabolism pathways and other pathways are compared between the control and Mito-HNK treated treatment groups.

Figure 4. Significant pathway changes associated with Mito-HNK treatment in lung adenoma cells. In lung adenoma cells, Mito-HNK treatment decreased the expression of genes associated with complex I, OXPHOS, glycolysis, and STAT3 signaling, while the treatment increased the expression of genes associated with apoptosis and cell death pathways. In addition, Mito-HNK treatment inhibited cysteine and methionine metabolism, fatty acid metabolism, the pentose phosphate pathway, and propanoate and the pyruvate metabolism, while the treatment hyperpolarized mitochondrial membrane potential in lung adenoma cells.

Figure 5. Clustering analysis based on scRNA-seq data of CD45− marker flow sorted mouse lung samples from control and Mito-LND treated mice. (A) Lung adenoma cells were identified using scCancer software. Marker gene expression for (B) epithelial cells, (C) endothelial cells, and (D) fibroblasts. (E) Annotation of CD45− cells into the four identified cell populations.

Figure 6. Pathway changes in the CD45− cell populations from control and Mito-LND treated groups. KEGG metabolism pathways and other pathways were compared between the control and Mito-LND treated groups.

Figure 7. Pathway changes associated with Mito-LND treatment in lung adenoma cells. In the lung adenoma cells, Mito-LND treatment significantly decreased the expression of genes associated with complex I, complex II, OXPHOS and AKT/ mTOR/p70S6K signaling, and increased the expression of genes associated with autophagy. In addition, Mito-LND treatment significantly decreased cysteine and methionine metabolism, while treatment hyperpolarized the mitochondrial membrane potential in lung adenoma cells.

Figure 8. Clustering analysis based on scRNA-seq data from the CD45− marker flow-sorted mouse lung samples of control and combined treatment groups. (A) Lung adenoma cells were identified based on the cell malignancy estimation. Marker gene expression for (B) epithelial cells, (C) endothelial cells, and (D) fibroblasts. (E) Annotation of the CD45− cells into the four identified cell populations.

Figure 9. Pathway alterations in CD45− cells from control vs combination treated mice. KEGG metabolism pathways and other pathways in CD45− mouse lung cells were compared between the control and combined treatment groups.

Figure 10. Significantly altered pathways in lung adenoma cells from mice given the combined treatment. In lung adenoma cells, the combined treatment decreased the activities of complex I, complex II, OXPHOS, glycolysis, AKT/mTOR/p70S6K signaling, STAT3 signaling, arginine and proline metabolism, cysteine and methionine metabolism, fatty acid metabolism, N-glycan biosynthesis, propanoate metabolism, and tryptophan metabolism. In contrast, combined treatment resulted in hyperpolarized mitochondrial membrane potential.