Journal: Blood Cancer Journal

Article Title: Bruton’s tyrosine kinase inhibition re-sensitizes multidrug-resistant DLBCL tumors driven by BCL10 gain-of-function mutants to venetoclax

doi: 10.1038/s41408-025-01214-y

Figure Lengend Snippet: A Heatmap of differentially expressed genes in RIVA cells containing vector or BCL10 mutants, (FC ≥ 1.5x up or down) . B Venn diagram of the number of differentially expressed genes in BCL10 mutants compared to vector. C Top 10 upregulated (adjusted p value ≤ 0.001) Jensen compartment complexes using genes upregulated with a FC ≥ 2. D The top overlapping Hallmark gene set pathways upregulated (FDR q-value ≤ 0.25) in BCL10 mutants. E Western blot of RIVA and HBL1 cells induced with doxycycline for 24 h and probed as indicated. F TNFα ELISA assay on cell supernatant from RIVA, HBL1, and U2932 cells containing vector or BCL10 mutants. G Boxplot of gene expression levels of TNFRSF13B and USP2 of untreated DLBCL cases comparing mutant versus wildtype BCL10 cases.

Article Snippet: To more fully assess impact of BCL10 mutations on treatment resistance, we interrogated the 960-compound TargetMol Epigenetic Library.

Techniques: Plasmid Preparation, Western Blot, Enzyme-linked Immunosorbent Assay, Gene Expression, Mutagenesis

Journal: Blood Cancer Journal

Article Title: Bruton’s tyrosine kinase inhibition re-sensitizes multidrug-resistant DLBCL tumors driven by BCL10 gain-of-function mutants to venetoclax

doi: 10.1038/s41408-025-01214-y

Figure Lengend Snippet: A Gene ontology analysis of genes upregulated in BCL10 mutant RIVA cells showing top 10 of 49 significantly upregulated ontologies (adjusted p < 0.001). Ontologies that were completely overlapping in genes were combined. B GSEA “BIOCARTA_CYTOKINE_PATHWAY” for BCL10 mutants (q = 0.082 S136X, q = 0.176 for R58Q). C Venn diagram of cytokine genes upregulated in BCL10 mutants with array validations indicated. D Boxplot of gene expression levels of CCL22 and IL7 of untreated DLBCL cases comparing mutant versus wildtype BCL10 cases. E ELISA assays of IL6, IL7 and TNFβ on cells supernatant of RIVA and HBL1 cells induced for 24 (IL7 and TNFβ) or 72 h (IL6) with doxycycline, ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: To more fully assess impact of BCL10 mutations on treatment resistance, we interrogated the 960-compound TargetMol Epigenetic Library.

Techniques: Mutagenesis, Gene Expression, Enzyme-linked Immunosorbent Assay

Journal: Blood Cancer Journal

Article Title: Bruton’s tyrosine kinase inhibition re-sensitizes multidrug-resistant DLBCL tumors driven by BCL10 gain-of-function mutants to venetoclax

doi: 10.1038/s41408-025-01214-y

Figure Lengend Snippet: A Genes that were upregulated (FC ≥ 2) in both mutants were analyzed for enrichment in regulation by transcription factors from the ENCODE project through the online transcription factor analysis tool ChIP-X Enrichment Analysis Version 3 (ChEA3). Sixteen transcription factors were significantly upregulated in BCL10 mutants (FDR ≤ 0.05). B Schematic of transcription factors identified to be upregulated in ( A ). C Boxplot of gene expression levels of BATF and IRF4 of untreated DLBCL cases comparing mutant versus wildtype BCL10 cases. D Western blot of RIVA and HBL1 cells induced with doxycycline for 24 h and probed as indicated. E Western blot of RIVA and U2932 cells induced with doxycycline for 24 h and probed as indicated. F ELISA assays in RIVA AND U2932 cell supernatant probing for CXCL10 (RIVA - S136X: p = 0.0046, R58Q: p = 0.0003, U2932- S136X: p = 0.0003, R58Q: p = 0.0028).

Article Snippet: To more fully assess impact of BCL10 mutations on treatment resistance, we interrogated the 960-compound TargetMol Epigenetic Library.

Techniques: Gene Expression, Mutagenesis, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Blood Cancer Journal

Article Title: Bruton’s tyrosine kinase inhibition re-sensitizes multidrug-resistant DLBCL tumors driven by BCL10 gain-of-function mutants to venetoclax

doi: 10.1038/s41408-025-01214-y

Figure Lengend Snippet: A Dose response viability assays of RIVA cells treated with pirtobrutinib (S136X: p = 0.033, R58Q: p = 0.019), acalabrutinib (S136X: p = 0.07, R58Q: p = 0.07), and ibrutinib (S136X: p = 0.023, R58Q: p = < 0.0001). B Epigenetic Library (TargetMol) on doxycycline induced RIVA cells. Hits are compounds that significantly inhibited cell viability and compounds that had an increase or decrease in viability two standard deviations from the mean of the difference between BCL10 S136X and vector after 72 h of 10 μM drug treatment. Targets in red highlight mutant-driven resistance, mutant-sensitive targets are in blue, and compounds that sensitized both vector and BCL10 S136X are in purple. C Schematic of hits from 4B. D Dose response viability assays of RIVA cells treated with selcidemstat (p=ns), tovorafenib (p = ns), ulixertinib (S136X: p = 0.0125, R58Q: p = 0.0103), JNKi (p = ns), duvelisib (S136X: p = 0.126, R58Q: p = 0.0264), idelalisib (S136X: p = ns, R58Q: p = 0.004), capivasertib (S136X: p = 0.0002, R58Q: p = 0.005), venetoclax (S136X: p = 0.008, R58Q: p = 0. 0.046), and AZD1208 (PIMi) (S136X: p = 0.019, R58Q: p = 0.040).

Article Snippet: To more fully assess impact of BCL10 mutations on treatment resistance, we interrogated the 960-compound TargetMol Epigenetic Library.

Techniques: Plasmid Preparation, Mutagenesis

Journal: Blood Cancer Journal

Article Title: Bruton’s tyrosine kinase inhibition re-sensitizes multidrug-resistant DLBCL tumors driven by BCL10 gain-of-function mutants to venetoclax

doi: 10.1038/s41408-025-01214-y

Figure Lengend Snippet: A Normalized counts from gene expression data for BCL2 family genes BCL2, BCL2L1 , and BCL2A1 in the BCL10 mutants. B Western blot of proteins implicated in venetoclax resistance including BCL-xL, BFL1, and BCL2 in doxycycline induced RIVA cells. C Transcript levels of doxycycline induced RIVA cells treated with DMSO or 5uM pirtobrutinib for 24 h: BCL2 (vector: p = 0.0325, R58Q: p = 0.005, S136X: p = 0.0006), BCL2L1 (vector: p=ns, R58Q: p = 0.0484, S136X: p = 0.0007), BCL2A1 (vector: p = 0.0087, R58Q: p = 0.0121, S136X: p = 0.0005). D Mitochondrial membrane potential flow cytometry assay in RIVA vector compared to RIVA R58Q and S136X at baseline. E Time course for BCL-xL, BFL1, and BCL2 in 5 μM pirtobrutinib treated RIVA cells. F Transcript levels of VDACs and TOMM22 after 24-h treatment with pirtobrutinib in RIVA cells. G Mitochondrial membrane potential flow cytometry assay in RIVA vector compared to RIVA R58Q and S136X after treatment with DMSO, pirtobrutinib, venetoclax or the combination for 2, 6 or 20 h. H Western blot of doxycycline induced RIVA cells treated with DMSO, venetoclax, pirtobrutinib or the combination for 24 h.

Article Snippet: To more fully assess impact of BCL10 mutations on treatment resistance, we interrogated the 960-compound TargetMol Epigenetic Library.

Techniques: Gene Expression, Western Blot, Plasmid Preparation, Membrane, Flow Cytometry

Journal: Blood Cancer Journal

Article Title: Bruton’s tyrosine kinase inhibition re-sensitizes multidrug-resistant DLBCL tumors driven by BCL10 gain-of-function mutants to venetoclax

doi: 10.1038/s41408-025-01214-y

Figure Lengend Snippet: A Upregulated transcription factor protein-protein interactions in genes upregulated by the BCL10 mutants probed through the Enrichr website ( p < 0.05). B Western blot showing phosphorylated STAT3 and total STAT3 in HBL1 and RIVA cells containing BCL10 mutants. C IL6 transcript levels of RIVA vector and mutants treated with pirtobrutinib for 24 h ( p < 0.0001). D IL6 cytokine levels in RIVA vector and mutants supernatant treated/untreated with 10uM pirtobrutinib for 24 h. E , F Western blot of doxycycline induced RIVA cells treated with DMSO, venetoclax, pirtobrutinib or the combination for 24 h. G Western Blot analysis of RIVA S136X cells treated with increasing concentrations of ruxolitinib with or without tocilizumab (IL6 inhibitor). H Western Blot analysis of RIVA S136X cells treated with increasing concentrations of ruxolitinib with or without the addition of TNFa. I IL6 transcript levels of RIVA S136X cells treated with TNFa alone or with ruxolitinib. J Schematic of TNFa / IL6/JAK/STAT signaling in BCL10 mutant cells. K Dose response viability assays of RIVA and HBL1 cells treated with pirtobrutinib (HBL1 clone1: p = 0.0027, HBL1 clone2: p = 0.0013, RIVA clone1: p = 0.0029, RIVA clone2: p < 0001) and ibrutinib (HBL1 clone1: p = 0.0401, HBL1 clone2: p = 0.0028, RIVA clone1: p = 0.3089, RIVA clone2: p < 0001), clones1 and 2 are the mutation (S136X) engineered at BCL10 gene locus. L Synergy assessments of venetoclax plus pirtobrutinib in endogenous BCL10 mutants (S136X) with reported synergy score.

Article Snippet: To more fully assess impact of BCL10 mutations on treatment resistance, we interrogated the 960-compound TargetMol Epigenetic Library.

Techniques: Protein-Protein interactions, Western Blot, Plasmid Preparation, Mutagenesis

Journal:

Article Title: Toll-like Receptor 4 Mediates Induction of the Bcl10-NF?B-Interleukin-8 Inflammatory Pathway by Carrageenan in Human Intestinal Epithelial Cells *

doi: 10.1074/jbc.M708833200

Figure Lengend Snippet: Effects of blocking antibody for TLR4 on CGN-induced increases in IL-8 and Bcl10. a, IL-8 was measured following exposure of NCM460 cells to λCGN (1 and 10 μg/ml for 24 h) following pretreatment with TLR4 blocking antibody (HTA-125) at concentrations of 10 and 20 μg/ml for 2 h. A dose-response effect is seen, with a reduced concentration of IL-8 in relation to a higher concentration of TLR4 blocking antibody. IL-8 in spent media was measured by ELISA and is expressed in pg/mg cell protein. Declines in IL-8 are statistically significant (*, p < 0.001). b, Bcl10 in the cell lysate was measured under conditions similar to those in A, with λCGN (1 μg/ml) and TLR4 antibody (10 μg/ml). An 80% decline in the Bcl10 protein occurred (***, p < 0.001).

Article Snippet: Immobilized Bcl10 molecules were detected by a mouse monoclonal antibody to Bcl10 (Novus Biologicals, Littleton, CO) and goat anti-mouse IgG-horseradish peroxidase complex (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Blocking Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Toll-like Receptor 4 Mediates Induction of the Bcl10-NF?B-Interleukin-8 Inflammatory Pathway by Carrageenan in Human Intestinal Epithelial Cells *

doi: 10.1074/jbc.M708833200

Figure Lengend Snippet: CGN-induced increase in Bcl10 and IL-8 is MyD88-dependent. a, MyD88 was silenced by insertion of a dominant-negative plasmid for MyD88. CGN-induced IL-8 increase declined 78% from 1274 ± 117 to 776 ± 41 pg/ml. This finding suggests that there is an alternative, minor pathway of CGN-induced IL-8 activation that is not mediated by TLR-4-Bcl10. b, similarly, following knockdown of MyD88, CGN-induced increase in Bl10, as determined by ELISA, was completely inhibited, declining from a peak of 3.83 ± 0.17 to a base line of 1.21 ± 0.08 (***, p < 0.001, one-way ANOVA with Tukey-Kramer post-test for multiple comparisons). Cn, control.

Article Snippet: Immobilized Bcl10 molecules were detected by a mouse monoclonal antibody to Bcl10 (Novus Biologicals, Littleton, CO) and goat anti-mouse IgG-horseradish peroxidase complex (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Dominant Negative Mutation, Plasmid Preparation, Activation Assay, Knockdown, Enzyme-linked Immunosorbent Assay, Control

Journal:

Article Title: Toll-like Receptor 4 Mediates Induction of the Bcl10-NF?B-Interleukin-8 Inflammatory Pathway by Carrageenan in Human Intestinal Epithelial Cells *

doi: 10.1074/jbc.M708833200

Figure Lengend Snippet: Inhibition of IRAK reduces CGN-associated increases in Bcl10 and IL-8. a, marked decline in IL-8 of the spent media follows treatment with IRAK1/4 inhibitor. b, similarly, a marked decline in Bcl10 protein content follows pretreatment with IRAK 1/4 inhibitor, placing Bcl10 downstream of IRAK in the cascade induced by CGN exposure. Cn, control; CGN, λCGN (1 μg/ml for 24 h); I, IRAK inhibitor. ***, p < 0.001 (one-way ANOVA with Tukey-Kramer post-test for multiple comparisons).

Article Snippet: Immobilized Bcl10 molecules were detected by a mouse monoclonal antibody to Bcl10 (Novus Biologicals, Littleton, CO) and goat anti-mouse IgG-horseradish peroxidase complex (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Inhibition, Control