byoral gavage with cabozantinib  (TargetMol)

Journal: Nature Communications

Article Title: The HDAC inhibitor romidepsin renders liver cancer vulnerable to RTK targeting and immunologically active

doi: 10.1038/s41467-025-62934-0

Figure Lengend Snippet: a Heatmap showing the gene expression values for all HDACs (the class is specified on the left) in 32 human HCC cell lines, analysing data at http://zucmanlab.com/ . b Heatmap showing the drug response of 32 human HCC cell lines to clinically relevant RTKi (orange) and epigenetic drugs (green), expressed as GI 50 , extracted from http://zucmanlab.com/ . c Dot plot reporting drug response expressed as GI 50 of the indicated human HCC cell lines to RTKi currently used in the clinic for HCC treatment (cabozantinib, lenvatinib, sorafenib, regorafenib). Data are presented as mean values +/- SEM. d Heatmap reporting cell viability after 48 h treatment with HDACi (romidepsin, EDOS-101, ACY957, panobinostat) and BETi (JQ1, OTX-015, mivebresib) at the indicated concentrations. Data are expressed as means of at least three independent experiments (scatter dot-plots are reported in Fig. with corresponding statistics). Viability percentages are reported using a blue (high)-to-red (low) colour code (the scale depicted on the right is used as a reference in all viability studies). e Western blots of the reported proteins in the indicated human HCC cell lines non-treated (NT) and treated with romidepsin (0.03 µM) for 8 h and 24 h. Quantifications are reported in Fig. with corresponding independent experiments and statistics. f Western blots displaying expression of the indicated proteins in JHH5 and HLE cells non-treated and treated with mivebresib (1 µM) for 8 h and 24 h (one independent experiment). Source data are provided as a Source Data file.

Article Snippet: Alb-R26 Met mice carrying liver tumours were treated every two days by oral gavage with cabozantinib (10 mg/kg; TargetMol) and every four days by intraperitoneal injection with romidepsin (1 mg/kg; TargetMol).

Techniques: Gene Expression, Western Blot, Expressing

Journal: Nature Communications

Article Title: The HDAC inhibitor romidepsin renders liver cancer vulnerable to RTK targeting and immunologically active

doi: 10.1038/s41467-025-62934-0

Figure Lengend Snippet: a Heatmap reporting effects on viability of cells exposed to romidepsin (Romi), cabozantinib (Cabo) or lenvatinib (Lenva), alone or in combination, at the indicated doses (µM) for 48 h. Cells were treated with the indicated inhibitors, alone and in combinations by adding the drugs at the same time (24 h after seeding). Data are the mean of three independent experiments. Scatter dot plots are shown in Fig. with corresponding statistics, two-tailed one-way ANOVA followed by Tukey multiple comparison; p < 0.0001 for all cell lines. b Synergy maps for the indicated drug combinations reported in ( a ). c Heatmap showing the effects on viability of cells following combined treatments of romidepsin with foretinib (Fore), regorafenib (Rego), or sorafenib (Sora) for 48 h. Data are the mean of three independent experiments (full heatmap including romidepsin and cabozantinib treatment is reported in Fig. , scatter dot plots are shown in Fig. with corresponding statistics, two-tailed one-way ANOVA followed by Tukey multiple comparison; p < 0.0001 for all cell lines). Significance: * ,§ : p < 0.05; ** ,§§ : p < 0.01; *** ,§§§ : p < 0.001. § indicates single treatments versus controls; * indicates combined treatments versus their respective single treatments. Source data are provided as a Source Data file.

Article Snippet: Alb-R26 Met mice carrying liver tumours were treated every two days by oral gavage with cabozantinib (10 mg/kg; TargetMol) and every four days by intraperitoneal injection with romidepsin (1 mg/kg; TargetMol).

Techniques: Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: The HDAC inhibitor romidepsin renders liver cancer vulnerable to RTK targeting and immunologically active

doi: 10.1038/s41467-025-62934-0

Figure Lengend Snippet: a Top panels: graphs reporting top-15 most significant enriched pathways according to GO_Biological_Process analyses in Huh7 cells treated with romidepsin (Romi), cabozantinib (Cabo), and RomiCabo versus non-treated cells (NT). Bottom panels: graphs reporting top-20 enrichments based on GO_Biological_Process analysis, with up-regulated (left) and down-regulated (right) pathways in the indicated conditions (RomiCabo versus cabozantinib-treated Huh7 cells). Pathways related to the mitotic spindle and lipid metabolism are highlighted in green and orange, respectively. Complete proteomics data are in Supplementary Data - . b Heatmap reporting z-score values of proteins related to fatty acids and triglycerides, cholesterol, and ceramides metabolic pathways in the indicated conditions (R: romidepsin; C: cabozantinib; R + C: RomiCabo). a, b A two-tailed heteroscedastic t-test was performed between each condition. Proteins with p values < 0.05 were considered as significant; n = 3 independent biological experiments. c, d Heatmap representation of unsupervised hierarchical clustering and principal component analysis (PCA) plots from lipidomic analysis of ceramides ( c ) and lysophospholipids ( d ) in Huh7 and Hep3B cells. e Triacylglycerols dot-plots for association of carbon atoms (fatty acid chain length) and double-bonds (saturation) from lipidomic analysis. Each lipid is represented by a circle and the colour of the circle correlates with the condition association direction (red: positive, blue: negative). The size of the circle correlates with the statistical significance (larger circles represent smaller p-values). c–e Multivariate (PCA, Hierarchical heatmap); n = 4 independent biological experiments. Source data are provided as a Source Data file.

Article Snippet: Alb-R26 Met mice carrying liver tumours were treated every two days by oral gavage with cabozantinib (10 mg/kg; TargetMol) and every four days by intraperitoneal injection with romidepsin (1 mg/kg; TargetMol).

Techniques: Two Tailed Test

Journal: Nature Communications

Article Title: The HDAC inhibitor romidepsin renders liver cancer vulnerable to RTK targeting and immunologically active

doi: 10.1038/s41467-025-62934-0

Figure Lengend Snippet: a Top-15 most significant enriched pathways according to IPA analyses from the proteome of Huh7 cells treated with romidepsin (Romi), cabozantinib (Cabo), or RomiCabo versus non-treated cells (NT). Pathways related to LXR/RXR and FXR/RXR are highlighted in blue. b z-score from the proteome for proteins related to LXR/RXR (green) or FXR/RXR (red); some proteins have overlapping upstream regulators LXR and FXR. ( a, b ) A two-tailed heteroscedastic t-test was performed between each condition. Protein with p values < 0.05 were considered as significant; n = 3 independent biological experiments. c Scheme illustrating the explored mechanism of action of romidepsin in relation to LXR/RXR and FXR/RXR pathways and lipid metabolism regulation. d LXR luciferase reporter assay in non-treated and treated Huh7 cells. Luciferase activity is reported as arbitrary units (a.u). An LXR agonist (GSK3987) was used as a positive control ( n = 3 independent biological experiments; p < 0.0001). e mRNA expression by RT-qPCR (expressed as RQ) of SREBF1 , MLXIPL , CD36 , and SCD1 in romidepsin (R), cabozantinib (C), or RomiCabo versus non-treated Huh7 cells. On the right of each graph are values corresponding to cells after treatment with FXR agonist (FXR agonist 3), LXR agonist (GSK3987), or LXR antagonist (GSK2033), either alone or in combination with romidepsin and RomiCabo (at least four independent experiments; SREBF1 , MLXIPL , CD36 , and SCD1 p < 0.0001). f, g mRNA expression by RT-qPCR of CYP7A1 ( n = 11 independent biological experiments, p = 0.0077) ( f ), ACACA ( n = 7 independent biological experiments, p = 0.0043), and FASN ( n = 4 independent biological experiments, p < 0.0001) ( g ) in the indicated conditions. h Representations of different transcriptional expression outcomes in LXR and/or FXR pathways after treatment with romidepsin. d –g two-tailed unpaired one-way ANOVA followed by Tukey’s ( d, e ) or Dunnet’s ( f, g ) multiple comparison. Significance: * ,$,#,%,£ p < 0.05; ** ,$$,##,%%,££ p < 0.01; ** *,$$$,###,%%%,£££ p < 0.001. * is comparison versus NT or as indicated by the statistical lines; $ versus romi; # versus cabo; % versus RomiCabo; £ versus FXR agonist+Romi. Schemes in c and h were performed with public open source Servier Medical Art, licensed under CC BY 4.0. Source data are provided as a Source Data file.

Article Snippet: Alb-R26 Met mice carrying liver tumours were treated every two days by oral gavage with cabozantinib (10 mg/kg; TargetMol) and every four days by intraperitoneal injection with romidepsin (1 mg/kg; TargetMol).

Techniques: Two Tailed Test, Luciferase, Reporter Assay, Activity Assay, Positive Control, Expressing, Quantitative RT-PCR, Comparison

Journal: Nature Communications

Article Title: The HDAC inhibitor romidepsin renders liver cancer vulnerable to RTK targeting and immunologically active

doi: 10.1038/s41467-025-62934-0

Figure Lengend Snippet: a Representative bright-field images of Huh7 cells non-treated (NT), treated with romidepsin (Romi), cabozantinib (Cabo), or RomiCabo for 24 h. White arrowheads indicate round cells. Percentage of round cells (normalized over total cell number, n = 3 independent biological experiments; p < 0.0001)). b Immunostaining of Huh7 cells after 24 h of treatment: Actin (cyan), β-Tubulin (magenta), Nuclei (yellow; n = 3 independent biological experiments). White arrows indicate round cells. c, d Immunostaining of monopolar and bipolar spindles with α-Tubulin (green), pS 10 H3 (red), nuclei (blue), ( n = 3 independent biological experiments; p < 0.0001). e Immunostaining of aberrant spindles with NuMa (yellow) and α-Tubulin (magenta; n = 3 independent biological experiments). f Maximal intensity projection (MIP) in XY plan of Huh7 cells treated for 24 h with romidepsin: α-Tubulin (magenta), pS 10 H3 (cyan), nuclei (yellow), reported as representative 3D reconstruction. g pS 10 H3 (S10) and pS 28 H3 (S28) positive cells at 24 h in non-treated (NT), romidepsin (R), cabozantinib (C), RomiCabo (R + C), and PLK1i conditions, normalised over total cell number (DAPI; n = 3 independent biological experiments; p < 0.0001). h Percentage of double-positive pS 10 H3 and pS 28 H3 alive cells after 16 h (left; n = 3 independent biological experiments; p = 0.0017) and 24 h (right; n = 3 independent biological experiments; p = 0.0017) of indicated treatments, measured by flow cytometry. i, j Representative cytometry profiles ( i ) of the DNA content (2n, S, 4n) of alive cells after 24 h, with Propidium Iodide and ( j ) quantification among alive cells ( n = 5 independent biological experiments; p < 0.0001). k Median fluorescence intensity (MFI) of γH2AX among live cells after 24 h by flow cytometry ( n = 6 independent biological experiments; p < 0.0001). l Western blots of reported signalling proteins (quantifications are shown in Figs. – , with the corresponding statistics, n = 3 independent biological experiments). Statistical analysis was performed with one-way ANOVA ( a, g ) or two-way ANOVA ( d ), followed by Tukey multiple comparison ( a, d, g ). Paired two-tailed Friedman followed by Dunn’s multiple comparison ( h, k ). Unpaired two-tailed two-way ANOVA followed by Dunnet’s multiple comparison ( j ). In ( g, i, j ) PLK1i condition was used as positive control of mitotic arrest. Levels of significance: * ,$,# p ≤ 0.05; ** ,$$,## p ≤ 0.01; *** ,$$$,### p ≤ 0.001. Bars represent SEM and notches in dot-plots represent mean levels. *indicates the significance between treated versus non-treated cells; $ is versus romidepsin; # is versus cabozantinib. Source data are provided as a Source Data file.

Article Snippet: Alb-R26 Met mice carrying liver tumours were treated every two days by oral gavage with cabozantinib (10 mg/kg; TargetMol) and every four days by intraperitoneal injection with romidepsin (1 mg/kg; TargetMol).

Techniques: Immunostaining, Flow Cytometry, Cytometry, Fluorescence, Western Blot, Comparison, Two Tailed Test, Positive Control

Journal: Nature Communications

Article Title: The HDAC inhibitor romidepsin renders liver cancer vulnerable to RTK targeting and immunologically active

doi: 10.1038/s41467-025-62934-0

Figure Lengend Snippet: a Viability of patient-derived tumoroids and liver organoids in romidepsin and RomiCabo treatments. The IC 50 of RomiCabo is shown, all others are included in Supplementary Table (p values in Supplementary Data ). Unpaired t-test corrected with Benjamini-Hochberg, stars compare Romi versus RomiCabo (red and blue: 0.68 µM and 1.3 µM cabozantinib, respectively; n ≥ 4 independent biological experiments). Tumoroid viability upon cabozantinib alone is reported in Fig. . b Protocol followed for in vivo study in Alb-R26 Met mice. ∅ indicate lesions of a mouse that died during “drug holiday”. The scheme was performed with free open source (pngegg.com). c Examples of PC-CT imaged tumours, with 3D reconstruction, before and after treatment: responding (left) and non-responding (right). Red squares: high magnification tumours. d, e Left: percentage of tumours according to their behaviour (evolutive, quiescent, regressive). Right: ratio between final versus initial tumour volume (logarithmic scale). NT: non-treated tumours. Tumours are considered regressive if this ratio is less than 0.85, evolutive if greater than 1.15, and quiescent between these two thresholds. Unpaired two-tailed Mann-Whitney ( d : p = 0.0013; e : p = 0.0041); number of independent samples is indicated. f Left: Sankey diagram represents lesions’ longitudinal behaviour at the end of the first session, of the “drug holiday”, and of the second session. Right: Ratio between final versus initial tumour volume (logarithmic scale) at the same time points as Sankey diagram. Unpaired two-tailed Mann-Whitney, p = 0.0061 (break) and p = 0.0025 (2 nd session), number of independent samples indicated in b, d, e . g–l Expression levels of class-I Hdacs ( g ), Rad23b ( h ), Mki-67 , Ccnd1 ( CyclinD1 ), Xiap , Survivin ( i ), Srebf1, Mlxipl ( j ), Fasn , Scd1 , Cd36 ( k ), Pd1, Pdl1 ( l ) in Alb-R26 Met non-treated (NT) versus RomiCabo treated (R + C) tumours. RT-qPCR (as RQ) relative to NT tumours. g–l Unpaired two-tailed Mann-Whitney, exact p values are indicated ( n ≥ 4 independent samples). Levels of significance: * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001. Bars represent SEM and notches in dot-plots represent mean levels. Source data are provided as a Source Data file.

Article Snippet: Alb-R26 Met mice carrying liver tumours were treated every two days by oral gavage with cabozantinib (10 mg/kg; TargetMol) and every four days by intraperitoneal injection with romidepsin (1 mg/kg; TargetMol).

Techniques: Derivative Assay, In Vivo, Two Tailed Test, MANN-WHITNEY, Expressing, Quantitative RT-PCR

Journal: Nature Communications

Article Title: The HDAC inhibitor romidepsin renders liver cancer vulnerable to RTK targeting and immunologically active

doi: 10.1038/s41467-025-62934-0

Figure Lengend Snippet: Romidepsin (Romi): 1 perturbs cell cycle signals and reduces cell survival regulators, 2 alters lipid metabolism modulators, and 3 causes defects in the mitotic machinery. Collectively, these events lead to a cytostatic effect and confer HCC cells dependency on RTK signalling support, which is an exploitable vulnerability. Consequently, the combination of romidepsin with RTKi (indicated here with cabozantinib; Cabo) leads to cytotoxic effects. Moreover, RomiCabo induces an immune infiltration and a switch towards an immune stimulatory profile, contributing to tumours regression. NT: non-treated cells. Scheme was performed with public open source Servier Medical Art, licensed under CC BY 4.0. Source data are provided as a Source Data file.

Article Snippet: Alb-R26 Met mice carrying liver tumours were treated every two days by oral gavage with cabozantinib (10 mg/kg; TargetMol) and every four days by intraperitoneal injection with romidepsin (1 mg/kg; TargetMol).

Techniques: