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. 2022 Feb 22;11:e71880. doi: 10.7554/eLife.71880

Figure 7. Homoharringtonine (HHT) has significant tumor killing activity both in vitro and in vivo.

(A) Results of best practice approach-based computational drevo as query signature. Top ranked 10 compounds with highest reversal potency were illustrated in the right panel. (B) Enrichment of HCC agents in compounds with reversal potency (XSum score<0). Statistical significance was determined based on the null distribution formed by 10,000 permutations. (C) 2D (left) and 3D (right) chemical structure of HHT. (D) Comparison of distribution of compound activity between HHT and three different drug categories, including chemotherapy (N=45 compounds), targeted cancer agents (N=419 compounds), and non-oncology (N=362 compounds). The IC50 values (from PRISM data set) of each drug category in each cell line (N=482) were determined through calculating the median IC50 value across all the compounds in this category. Data are presented as median±quartiles, N≥100. (E) The drug sensitivity data of HHT (achieved from PRISM data set) across liver cancer cell lines. The drug sensitivities of two HCC agents in the first-line (sorafenib and lenvatinib) and one HCC agent in the second-line (regorafenib) were also presented for comparison. Areas with different colors denote the interquartile range of median IC50 values of compounds within different drug categories. (F) Long-term cell proliferation assay for testing the anti-tumor activity of HHT across 10 liver cancer cell lines. Of these, four cell lines have not been profiled by PRISM for the sensitivity to HHT. (G) Macroscopic image of tumors harvested from xenograft mice treated with vehicle (upper) and HHT (lower). (H) Longitudinal tumor volume progression of subcutaneous MHCC97H xenograft tumors treated with vehicle (N=6) and HHT (N=6). The statistical significance of difference between groups was determined using Student’s t-test. Data are represented as mean ± SD. (I) Body weight changes of mice in control (N=6) and HHT-treated (N=6) groups. Statistical significance was determined using Student’s t-test. Data are represented as mean ± SD. *p<0.05, **p<0.01, ***p<0.001. NS, not significant. HCC, hepatocellular carcinoma.

Figure 7—source data 1. Drug-induced expression changes across different cell lines as well as different concentrations.

Figure 7.

Figure 7—figure supplement 1. Reversal effect of HHT on Sigevo across different conditions.

Figure 7—figure supplement 1.

(A) Schematic figure illustrating the experimental design. (B) Enrichment scores of ascending (upper) and descending (lower) gene sets in the condition of different cell types as well as different concentrations. A positive enrichment score of a given gene set indicates the enrichment of HHT-induced upregulated genes and a negative enrichment score indicates the enrichment of HHT-induced downregulated genes. Color toward gray indicates no statistical significance. (C) GSEA plots of ascending and descending gene sets in different conditions. HHT, homoharringtonine.
Figure 7—figure supplement 2. Clinical and biological characterization of RPL3 in liver cancer.

Figure 7—figure supplement 2.

(A) The comparison of mRNA expression level of RPL3 between tumor and non-tumor tissues across seven HCC clinical cohorts. Statistical significance of difference was determined using Wilcoxon rank-sum test. Color toward gray indicates no statistical significance. (B) Representative images of immunohistochemical staining of RPL3 in liver normal (left) and tumor tissues (right) from the Human Protein Atlas (HPA) program. (C) Comparison of survival curves between high RPL3 expression and low RPL3 expression groups. (D) Distribution of gene dependency score (CERES score) of 18,333 protein-coding genes in liver cancer cell lines. A lower CERES score of certain gene indicates a higher likelihood that this gene is essential in cell growth and survival. (E) The gene dependency of RPL3 across 22 liver cancer cell lines.
Figure 7—figure supplement 3. The effect of HHT on cell proliferation across 10 liver cancer cell lines.

Figure 7—figure supplement 3.

Cell proliferation rate was analyzed by IncuCyte ZOOM system every 4 hr for 100 hr. HHT, homoharringtonine.
Figure 7—figure supplement 4. Summary of the anti-tumor effect of HHT-sorafenib combination across 10 liver cancer cell lines.

Figure 7—figure supplement 4.

Heatmap represents the summary of the synergistic effects of HHT-sorafenib combination. Higher score indicates greater synergism. The lower panel displays two representative surface plots that show the overall synergistic (left) and antagonistic (right) effects of the combination treatments. HHT, homoharringtonine.
Figure 7—figure supplement 5. Detailed information of the anti-tumor effect of HHT-sorafenib combination across 10 liver cancer cell lines.

Figure 7—figure supplement 5.

Combinational effects were measured using three models, including Bliss independence model, Loewe additivity model, and Highest single agent (HAS) model. HHT, homoharringtonine.