Combinatorial treatment with IDX and cis inhibits tumor growth along with increasing T cells infiltration.
(A) Representative mice and dissected tumors. For one mouse from the cis group, where the inoculated tumor was deeply located inside the organ tissue, that tumor could not be clearly dissected out and therefore was not its representative image is not shown. (B) Tumor volume during treatment. Tumor xenografts were established in athymic nude mice by inoculation of C17 cancer cells. After the tumors had grown to approximately 50 mm3, the animals were randomly divided into 6 groups (5 mice/group), which were treated every-other-day with 5 mg/kg cis, 10 mg/kg IDX, 20 mg/kg, 5 mg/kg cis plus 10 mg/kg IDX or 5 mg/kg cis plus 20 mg/kg IDX. Control mice received saline. Tumors were measured each other day over a total of 21 days after cancer cell inoculation and tumor volume was calculated. * p≤0.05 and ***p<0.0001 when compared to control group. (C) Violin plot showing the percentage of infiltrating cells expressing CD14+ (monocyte) and CD19+ (B cells) in total live lymphocytes and cells expressing CD4+, CD8+ and CD4+ CD8+in CD19- populations isolated from mice tumor, measured by flow cytometry. Each point represents one mouse in each group. ** p≤0.01 when compared to cis group. (D) Flow cytometric analysis on tumor infiltrating CD8 + T cells upon aforementioned treatment in total live lymphocytes. * p≤0.05 when compared to DMSO group. (E) Expression of ENOX2 at single-cell resolution from a publicly available sequencing cohort GSE150430. UMAP plot generated by using all cells harvested from 15 patients. Epithelial clusters enriched with mainly cells from tumor were circled and found to be the major cell type expressing ENOX2. (F) UMAP plot of malignant cells from 11 patients. Some patients including patient 11 and 13 were found to contain malignant cells with high expression of ENOX2. (G) Representative tumor histology staining of NPC clinical tissues for ENOX2 expression. (H) Analysis of ENOX2 gene expression and methylation from four independent public microarray datasets GSE34573, GSE53819, GSE12452 and GSE52068. * p≤0.05 when compared to corresponding normal tissue.