Histone deacetylases 10 as a prognostic biomarker correlates with tumor microenvironment and therapy response in colorectal cancer

Figure 1 Higher histone deacetylases 10 expression in colorectal cancer and was associated with poor prognosis. A: Higher histone deacetylases 10 (HDAC10) expression in normal and tumor tissues in pan-cancer data from the TIMER website; B: HDAC10 expression analysis in paired colorectal cancer (CRC) samples from TCGA-COAD showed high expression in tumor; C: Quantitative real-time polymerase chain reaction (qRT-PCR) analysis from thirty patients tissues indicated HDAC10 upregulated in CRC; D: Representative cases in normal tissues and CRC showed high HDAC10 expression in CRC. Scale bar, 100 μm and 20 μm; E: Quantification of immunohistochemistry (IHC) positive areas in normal and CRC indicated high levels of HDAC10 in CRC; F: Receiver operating characteristic curve based on HDAC10 for predicting 1-, 3- , 5-year overall survival; G: Kaplan-Meier survival analysis of HDAC10 expression in CRC tissue microarray and TCGA-COAD datasets showed patients with expressing higher levels of HDAC10 had poor prognosis. ^aP < 0.001, ^bP < 0.01, ^cP < 0.05 vs the NC group. The statistical power for comparing the rates between NC group and CRC group in qRT-PCR and IHC experiments using PASS software, and the results were 71% and 73%, respectively. CRC: Colorectal cancer; OS: Overall survival; TCGA: The Cancer Genome Atlas; HDAC10: Histone deacetylases 10; AUC: Area under the curve.

Figure 2 Histone deacetylases 10 was associated with clinicopathological characteristics and nomogram model construction. A: Differential analysis for histone deacetylases 10 (HDAC10) expression in patients with different age; B: Gender; C: High HDAC10 expression in high grade; D: Status; E: High levels of HDAC10 in advanced T stage; F: High HDAC10 expression in advanced N stage; G: High HDAC10 expression in advanced M stage; H: High levels of HDAC10 in advanced pathologic stage; I and J: Univariate Cox and multivariate Cox regression analysis for clinical factors and HDAC10 expression with overall survival (OS). Hazard ratio > 1 represented risk factors for survival and hazard ratio < 1 represented protective factors; K: Nomogram for predicting the 1-, 3-, and 5-year OS from CRC tissue microarray; L: Calibration curves for predicting 1-, 3-, and 5-year OS; M: Receiver operating characteristic curve for predicting 1-, 3-, and 5-year OS; N: Decision curve analysis based on nomogram better determine survival than only HDAC10. OS: Overall survival; HDAC10: Histone deacetylases 10; AUC: Area under the curve.

Figure 3 Histone deacetylases 10 promote tumor proliferation and migration. A: Knockdown of histone deacetylases 10 (HDAC10) expression in SW480 and HCT116 cells at mRNA and protein levels; B: Cell counting kit-8 assay showed HDAC10 depletion inhibited cell proliferation; C: Colony formation analysis indicated HDAC10 knockdown suppressed colony formation; D: Wound-healing assay showed HDAC10 depletion inhibited cell migration. Compared with the siControl group, ^aP < 0.001. Data were represented as mean ± SD of biological triplicates. HDAC10: Histone deacetylases 10.

Figure 4 Differential analysis and functional enrichment analysis. A: Volcano plot from the top five upregulated and downregulated differentially expressed genes (DEGs) between high and low-histone deacetylases 10 group; B: Heat map showed the top five genes in both upregulated and downregulated DEGs; C: Different expression levels of DEGs between normal colon and colorectal cancer; D: Gene set variation analysis showed the activation status of biological pathways in two groups. HDAC10: Histone deacetylases 10.

Figure 5 Correlation between histone deacetylases 10 and tumor microenvironment in The Cancer Genome Atlas database. A: Low expression of 4 immunomodulators (MHC, immunostimulators, chemokines and receptors) in high and low-histone deacetylases 10 (HDAC10) groups; B: Low stromal, immune, and ESTIMATE scores in high HDAC10 groups; C: HDAC10 expression positively correlated with tumor purity; D: Low infiltration level of common immune cells in high HDAC10 group; E: Correlation between HDAC10 expression and infiltration levels of tumor-infiltrating immune cells calculated by seven independent algorithms; F: Immunohistochemistry analysis showed that low expression of CD8a, CXCL9 and CXCL10 in high HDAC10 group. Scale bar, 100 μm; G: HDAC10 expression was negatively associated with critical steps in cancer-immunity cycle. ^aP < 0.001, ^bP < 0.01, ^cP < 0.05. NS: No significance; HDAC10: Histone deacetylases 10.

Figure 6 Effect on immune checkpoints and tumor burden mutation. A: Low expression of immune checkpoints in high histone deacetylases 10 (HDAC10) group; B: HDAC10 expression negatively correlated with PD-L1 in colorectal cancer tissue microarray; C: Low tumor burden mutation score in high HDAC10 group; D: Waterfall plot for somatic mutations in different HDAC10 groups. ^aP < 0.001, ^bP < 0.01, ^cP < 0.05. HDAC10: Histone deacetylases 10.

Figure 7 Immunotherapy and drug sensitivity analysis. A-H: Low immunotherapy response in high histone deacetylases 10 (HDAC10) group from Gide 2019, GSE53127, GSE91061, GSE100797, GSE103668, GSE111636, GSE126044, and IMvigor210 cohorts; I-P: Low common chemotherapy drug sensitivity in high HDAC10 group. HDAC10: Histone deacetylases 10.