Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Ibrutinib and PD-1 blockade potentiate mesothelin-targeting CAR-T cell therapy in preclinical models of pancreatic cancer

doi: 10.1158/1078-0432.CCR-25-2907

Figure Lengend Snippet: A) BTK expression as assessed by western blot analysis in the indicated pancreatic cancer cell lines or in the control Raji B cell lymphoma. B) Schematic of the experimental design. AsPC-1 tumor-bearing mice received ibrutinib treatment by oral gavage (25mg/kg) starting at the time of CAR-T cell transfer (1.5e6 cells) and up to day 35. C) Tumor growth kinetic (tumor volume, mean ± SEM) in AsPC-1 tumor-bearing mice treated with UTD T cells or 1.5e6 meso-CAR-T cells either as monotherapy or in combination with ibrutinib (treatment window is indicated by the green square; n=5–6 mice per group). Two-way ANOVA. HD207 was used to generate meso-CAR-T cells (transduction: 43%, viability at infusion: 84%). D) Tumor volume in each individual mouse from C. E) Absolute numbers (mean ± SEM) of meso-CAR-T cells in the blood of AsPC-1 tumor-bearing mice from C, treated as indicated and as shown in B. Two-way ANOVA. F) Expression of PD-1, TIM3 and CD39 markers (frequency, mean ± SEM) overtime on the surface of CD4-positive and CD8-positive CAR-T cells in the blood of tumor-bearing mice treated with CAR-T cells alone or in combination with ibrutinib treatment. G) Phenotype of CD4-positive and CD8-positive CAR-T cells in the peripheral blood of tumor-bearing mice at the indicated days post adoptive transfer of CAR-T cells alone or in combination with ibrutinib treatment. T stem cell memory cells (TSCM, CD45RA+CCR7+CD95+), central memory (CM, CD45RA-CCR7+), effector memory (CD45RA-CCR7-) and terminally differentiated effector memory (TEMRA, CD45RA+CCR7-) T cells. Unpaired Student’s t test. H) Tumor growth kinetic (tumor volume, mean ± SEM) in AsPC-1 tumor-bearing mice treated with UTD T cells or 1.5e6 meso-CAR-T cells either as monotherapy or in combination with ibrutinib (treatment window is indicated by the green square; n=5–6 mice per group). Two-way ANOVA. HD53 was used to generate meso-CAR-T cells (transduction: 72%, viability at infusion: 85%). I) Tumor volume in each individual mouse from H. I) Absolute numbers (mean ± SEM) of meso-CAR-T cells in the blood of AsPC-1 tumor-bearing mice from H, treated as indicated and as shown in B. Two-way ANOVA. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: All Q-PCR analyses were done using TaqMan probes from Applied Biosystems: Btk (Hs00975865_m1; FAM-MGB); Beta-actin (Hs99999903_m1, VIC-MGB).

Techniques: Expressing, Western Blot, Control, Transduction, Adoptive Transfer Assay

Journal: Frontiers in Oncology

Article Title: Combined cellular and biochemical profiling of Bruton’s tyrosine kinase inhibitor nemtabrutinib reveals potential application in MAPK-driven cancers

doi: 10.3389/fonc.2025.1667291

Figure Lengend Snippet: Biochemical kinase profiling. (a) Biochemical IC 50 values of nemtabrutinib and four approved BTK inhibitors on BTK. (b) Phylogenetic tree of human protein kinases highlighting 256 wild-type kinases that were examined for inhibition by nemtabrutinib in profiling experiments. Enzyme assays were performed at K M,bin ATP and 1 µmol/L nemtabrutinib. (c) Radar chart of the percentage inhibition of 254 wild-type kinases in the presence of 1 µmol/L nemtabrutinib or one of the four approved BTK inhibitors. Each dot represents a kinase and the kinases are ordered based on percentage inhibition per inhibitor.

Article Snippet: Nemtabrutinib and the approved BTK inhibitors were profiled on a panel of 254 wild-type kinases in mobility shift assays (MSA) at Carna Biosciences, Inc. (Kobe, Japan).

Techniques: Inhibition

Journal: Frontiers in Oncology

Article Title: Combined cellular and biochemical profiling of Bruton’s tyrosine kinase inhibitor nemtabrutinib reveals potential application in MAPK-driven cancers

doi: 10.3389/fonc.2025.1667291

Figure Lengend Snippet: Cancer cell panel profiling of nemtabrutinib. (a) Waterfall plot of IC 50 values of nemtabrutinib in cell viability assays with 160 cancer cell lines. (b) Dose-response curves of nemtabrutinib and the four approved BTK inhibitors in viability assays with SU-DHL-6 and REC-1 cells.

Article Snippet: Nemtabrutinib and the approved BTK inhibitors were profiled on a panel of 254 wild-type kinases in mobility shift assays (MSA) at Carna Biosciences, Inc. (Kobe, Japan).

Techniques:

Journal: Frontiers in Oncology

Article Title: Combined cellular and biochemical profiling of Bruton’s tyrosine kinase inhibitor nemtabrutinib reveals potential application in MAPK-driven cancers

doi: 10.3389/fonc.2025.1667291

Figure Lengend Snippet: Cancer gene mutation analysis and comparative profiling. (a) Volcano plot showing the correlation of nemtabrutinib response in cell viability assays with the mutation status of 23 oncogenic kinases in the cell lines. Each circle represents a kinase gene that is mutated in at least three cell lines. (b) Network tree connecting inhibitors with a similar profile. In case the Pearson correlation coefficient of the IC 50 fingerprint of two compounds is 0.5 or higher, a connection is drawn in the network. The length of the line has no meaning. Nemtabrutinib and the BTK inhibitors are colored in red.

Article Snippet: Nemtabrutinib and the approved BTK inhibitors were profiled on a panel of 254 wild-type kinases in mobility shift assays (MSA) at Carna Biosciences, Inc. (Kobe, Japan).

Techniques: Mutagenesis

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: BTK autoinhibition analyzed by high-throughput swaps of SH2 domains

doi: 10.1073/pnas.2502688122

Figure Lengend Snippet: SH2-domain substitutions increase fitness in BTK. Fitness scores for the 188 SH2-domain chimeras corresponding to Tec kinases or ancestral SH2 domains are shown. SH2 domains are ordered by their sequence relationships. The fitness bars, measured using the Jurkat assay, are colored by the human Tec kinase to which they are most closely related, requiring at least 75% sequence identity to display that color (sequences without 75% sequence identity to a human Tec kinase are colored purple). Error bars represent SEM and points are the individual replicate values. Sequence numbering corresponds to SI Appendix , Table S1 .

Article Snippet: The assembled BTK sequence was digested with BamHI HF (New England Biolabs) and XmaI (New England Biolabs) in a 30 μL reaction containing 3 μL of CutSmart buffer (New England Biolabs).

Techniques: Sequencing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: BTK autoinhibition analyzed by high-throughput swaps of SH2 domains

doi: 10.1073/pnas.2502688122

Figure Lengend Snippet: Screening αI kinase substitutions. The 118 αI kinase sequences are shown along with a multiple sequence alignment and fitness scores. The fitness scores are shown for the human BTK SH2 genetic background, the BMX-H SH2 genetic background, and the difference between these two backgrounds (BTK values subtracted from the BMX-H values). Error bars represent SEM and points represent the individual replicate values. For the difference values, error bars are determined using the SE propagation formula.

Article Snippet: The assembled BTK sequence was digested with BamHI HF (New England Biolabs) and XmaI (New England Biolabs) in a 30 μL reaction containing 3 μL of CutSmart buffer (New England Biolabs).

Techniques: Sequencing