anti tumor traditional chinese medicines  (TargetMol)

Journal: Journal for Immunotherapy of Cancer

Article Title: HDAC and MEK inhibition synergistically suppresses HOXC6 and enhances PD-1 blockade efficacy in BRAFV600E-mutant microsatellite stable colorectal cancer

doi: 10.1136/jitc-2024-010460

Figure Lengend Snippet: Drug screening results in BRAF V600E MSS CRC based on PDOs and cell lines. ( A ) Graphical overview of the study. ( B ) Heatmap of drug inhibition for the two PDOs. ( C ) Venn diagram showing the intersection of drugs with over 60% inhibition on the two PDOs. ( D ) Bar graph detailing specific inhibition rates of common drugs for the two PDOs. ( E ) Heatmap of drug responses on the three cell lines. ( F ) Venn diagram showing the intersection of drugs with over 60% inhibition on the three cells. ( G ) Bar graph showing specific inhibition rates of common drugs across the three cells. ( H ) Venn diagram showing the intersection of common drugs between the two PDOs and the three cell lines. BRAF, B-Raf proto-oncogene, serine/threonine kinase; CRC, colorectal cancer; HDAC, histone deacetylase; IHC, immunohistochemistry; MEK, mitogen-activated protein kinase kinase; MSS, microsatellite stable; PDO, patient-derived organoid; PD-1, programmed death 1; RNA-seq, RNA sequencing.

Article Snippet: To identify potential therapeutic agents for BRAF V600E MSS CRC, we intersected the Anti-cancer Compound Library (L3000, Selleck) and the Food and Drug Administration-approved Drug Library (L1300, Selleck), resulting in a custom subset of 768 drugs for screening.

Techniques: Drug discovery, Inhibition, Histone Deacetylase Assay, Immunohistochemistry, Derivative Assay, RNA Sequencing

Journal: Journal for Immunotherapy of Cancer

Article Title: HDAC and MEK inhibition synergistically suppresses HOXC6 and enhances PD-1 blockade efficacy in BRAFV600E-mutant microsatellite stable colorectal cancer

doi: 10.1136/jitc-2024-010460

Figure Lengend Snippet: HOXC6 contributes to treatment resistance in BRAF V600E MSS CRC and regulates the MAPK and AKT pathways and the MYC gene. ( A ) Left: HOXC6 expression in BRAF V600E and BRAF wild-type groups in TCGA COAD. ****p<0.0001 (Mann-Whitney U test). Right: HOXC6 expression in BRAF V600E dMMR, BRAF wild-type dMMR, BRAF V600E pMMR, and BRAF wild-type pMMR groups in TCGA COAD. ns, not significant; ***p<0.001 (Mann-Whitney U test). ( B ) HOXC6 expression in BRAF V600E MSS and BRAF wild-type MSS CRC cell lines from the CCLE database. **p<0.01 (Student’s t-test). ( C ) HOXC6 mRNA expression in HT-29 and HT-29_EnR cells. *p<0.05 (Student’s t-test). ( D ) IC50 values of encorafenib in HOXC6-knockdown HT-29_EnR cells and control cells. ***p<0.001 (Student’s t-test). ( E ) IC50 values of dabrafenib, encorafenib, and trametinib in HOXC6-overexpressing HT29 cells (HT29-HOXC6) and control cells (HT29-Control), respectively. Results represent means±SD (n=4). *p<0.05 (Student’s t-test). ( F ) Top 10 enriched pathways in HOXC6-overexpressing cells revealed by KEGG analysis. ( G ) Western blotting for HOXC6-overexpressing cells and control cells. ( H ) Western blotting for HOXC6-knockdown cells and control cells. ( I ) Cell viability profiles of four BRAF V600E MSS CRC cell lines treated with dabrafenib, encorafenib, and trametinib, respectively. Results represent means±SD (n=4). ( J ) Western blotting for the four BRAF V600E MSS CRC cell lines. ( K ) Peak annotation analysis indicating the distribution of binding sites. ( L ) Chromatin immunoprecipitation-quantitative results on the percentage of Input for MYC promoter. **p<0.01 (Student’s t-test). ( M ) Relative luciferase activity analyzed in the dual luciferase reporter assay to examine HOXC6 binding to the MYC promoter. MYC_WT, MYC_wild-type; MYC_Mut, MYC_mutant. Results represent means±SD (n=3). **p<0.01; ns, not significant (Student’s t-test). ( N ) Schematic of HOXC6 binding to the MYC gene. AKT, protein kinase B; BRAF, B-Raf proto-oncogene, serine/threonine kinase; COAD, colon adenocarcinoma; CRC, colorectal cancer; dMMR, deficient mismatch repair; ERK, extracellular signal-regulated kinase; HOXC6, homeobox C6; IC50, half-maximal inhibitory concentration; KEGG, Kyoto Encyclopedia of Genes and Genomes; MAPK, mitogen-activated protein kinase; MSS, mismatch repair; p-AKT, phosphorylated-AKT; p-ERK, phosphorylated-ERK; PI3K, phosphoinositide 3-kinase; pMMR, proficient mismatch repair; TCGA,The Cancer Genome Atlas; TPM, transcripts per kilobase million.

Article Snippet: To identify potential therapeutic agents for BRAF V600E MSS CRC, we intersected the Anti-cancer Compound Library (L3000, Selleck) and the Food and Drug Administration-approved Drug Library (L1300, Selleck), resulting in a custom subset of 768 drugs for screening.

Techniques: Expressing, MANN-WHITNEY, Knockdown, Control, Western Blot, Binding Assay, Chromatin Immunoprecipitation, Luciferase, Activity Assay, Reporter Assay, Mutagenesis, Concentration Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: HDAC and MEK inhibition synergistically suppresses HOXC6 and enhances PD-1 blockade efficacy in BRAFV600E-mutant microsatellite stable colorectal cancer

doi: 10.1136/jitc-2024-010460

Figure Lengend Snippet: Immune microenvironment characteristics and immune cell infiltration differ between BRAF V600E MSS CRC and BRAF wild-type MSS CRC. ( A ) Volcano plot demonstrating differentially expressed genes between patients with BRAF V600E MSS CRC and patients with BRAF wild-type MSS CRC. ( B ) Top 10 significantly enriched pathways in BRAF V600E MSS CRC identified by GO enrichment analysis. ( C )-( F ) GSEA analysis revealing significant enrichment in pathways such as immunoregulatory interactions between a lymphoid and a non-lymphoid cell ( C ), MHC class II antigen presentation ( D ), Hallmark_IL-2_STAT5_signaling ( E ), and Hallmark_IL-6_JAK_STAT3_signaling ( F ) in BRAF V600E MSS CRC. ( G )-( H ) GSEA analysis illustrating significant enrichment of IL-17 signaling pathway ( G ) and IL-10 signaling pathway ( H ) in BRAF V600E MSS CRC cell lines. ( I )- ( J ) Representative images of multiplex fluorescent immunohistochemistry for CD8 + cells, PD-1 + cells, and DAPI in BRAF wild ( I ) and BRAF V600E ( J ) MSS CRC tissues. Scale bars, 200 µm. ( K ) Quantification of PD-1 + cells in BRAF V600E MSS CRC and BRAF wild-type MSS CRC groups. *p<0.05 (Student’s t-test). ( L ) Quantification of CD8 + cells in BRAF V600E MSS CRC and BRAF wild-type MSS CRC groups. ns, not significant (Student’s t-test). ( M ) ssGSEA analysis showing the scores of immune cell infiltration in BRAF V600E MSS CRC and BRAF wild-type MSS CRC groups. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001 (Mann-Whitney U test). BRAF, B-Raf proto-oncogene, serine/threonine kinase; CRC, colorectal cancer; GO, Gene Ontology; GSEA, gene set enrichment analysis; IL, interleukin; KEGG, Kyoto Encyclopedia of Genes and Genomes; MHC, major histocompatibility complex; MSS, microsatellite stable; PD-1, programmed death 1; ssGSEA, single sample gene set enrichment analysis.

Article Snippet: To identify potential therapeutic agents for BRAF V600E MSS CRC, we intersected the Anti-cancer Compound Library (L3000, Selleck) and the Food and Drug Administration-approved Drug Library (L1300, Selleck), resulting in a custom subset of 768 drugs for screening.

Techniques: Immunopeptidomics, Multiplex Assay, Immunohistochemistry, MANN-WHITNEY

Journal: Journal for Immunotherapy of Cancer

Article Title: HDAC and MEK inhibition synergistically suppresses HOXC6 and enhances PD-1 blockade efficacy in BRAFV600E-mutant microsatellite stable colorectal cancer

doi: 10.1136/jitc-2024-010460

Figure Lengend Snippet: The characteristics of CT26 BRAF V637E cell line. ( A ) Western blotting for CT26 cell, CT26-Control cell, and CT26 BRAF V637E cell. ( B ) Tumor formation rates of subcutaneous inoculation of CT26-Control cells and CT26 BRAF V637E cells. ( C ) Inhibition rates of CT26 BRAF V637E cells under different treatments (trametinib, 0.1 µM; panobinostat, 0.3 µM). Results represent means±SD (n=3). **p<0.01 (Student’s t-test). ( D )-( F ) Gene expression changes of Ifng ( D ), Cxcl9 ( E ) and H2-T3 ( F ) in CT26 BRAF V637E cells under different treatments (trametinib, 0.1 µM; panobinostat, 0.3 µM), respectively. Results represent means±SD (n=3). *p<0.05; **p<0.01; ***p<0.001 (Student’s t-test). ( G ) Western blotting for the cGAS/STING pathway in CT26 BRAF V637E cells with different treatments (trametinib, 0.1 µM; panobinostat, 0.3 µM). ( H ) Western blotting for MMR proteins in CT26 BRAF V637E cells with different treatments (trametinib, 0.1 µM; panobinostat, 0.3 µM). ( I ) Cytokine arrays evaluating changes in the levels of cytokines secreted by CT26 BRAF V637E cells before and after the combination treatment (trametinib, 0.1 µM; panobinostat, 0.3 µM). BRAF, B-Raf proto-oncogene, serine/threonine kinase; cGAS/STING, cyclic guanosine monophosphate–adenosine monophosphate synthase/stimulator of interferon genes; ERK, extracellular signal-regulated kinase; Pano, panobinostat; p-ERK, phosphorylated-ERK; Tra, trametinib.

Article Snippet: To identify potential therapeutic agents for BRAF V600E MSS CRC, we intersected the Anti-cancer Compound Library (L3000, Selleck) and the Food and Drug Administration-approved Drug Library (L1300, Selleck), resulting in a custom subset of 768 drugs for screening.

Techniques: Western Blot, Control, Inhibition, Gene Expression