Journal: Molecular Biomedicine

Article Title: Curcumin analogue FLLL32 induces ferroptosis by targeting the STAT3-GPX4/SLC40A1 axis in colorectal cancer

doi: 10.1186/s43556-026-00575-5

Figure Lengend Snippet: FLLL32 suppresses CRC xenograft growth and inhibits the STAT3-GPX4/SLC40A1 axis in vivo . a Images of xenograft tumors from vehicle- and FLLL32-treated mice ( n = 6). b Tumor growth curves of HCT116 xenografts treated with vehicle or FLLL32 ( n = 6). c Statistics of tumor weight ( n = 6). d - f Ki67, pSTAT3 and GPX4 immunohistochemical staining images and H-score quantification in xenograft tumors ( n = 3). Scale bars = 25 μm. g Western blot analysis and quantification of STAT3, pSTAT3, GPX4, and SLC40A1 in xenograft tumor tissues. β-Tubulin was used as the loading control ( n = 3). h Body weight changes during vehicle or FLLL32 treatment. i Representative H&E staining images of major organs, including heart, spleen, liver, kidney, lung from vehicle- and FLLL32-treated mice. Scale bars = 50 μm. Data are presented as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: The sections were then incubated with primary antibodies against Ki67 (Proteintech, 27309-1-AP), Phospho-STAT3 (Tyr705) (CST, 9145), or GPX4 (HUABIO, ET1706-45) at 4 °C overnight.

Techniques: Xenograft Assay, In Vivo, Immunohistochemical staining, Staining, Western Blot, Analysis, Control

Journal: Journal for Immunotherapy of Cancer

Article Title: Engineering T cell-engaging bispecific antibodies targeting KRAS G12V neoantigen across HLA-A*11:01 and HLA-A*03:01

doi: 10.1136/jitc-2026-016411

Figure Lengend Snippet: Characterization of KRAS G12V-BsAb. ( A–B ) Selective binding of the purified scDb2 to KRAS G12V/HLA-A*11:01 ( A ) and CD3 ( B ) by ELISA. Biotinylated KRAS G12V/HLA-A*11:01 or recombinant CD3ε/δ was coated on a streptavidin plate and then incubated with scDb2 at 0.1, 1 or 10 nM, followed by detection with an anti-His antibody. Optical density at 450 nm was measured on a spectrophotometric plate reader. Data represent mean±SD of three technical replicates. ( C ) Surface plasmon resonance analysis of scDb2 binding to KRAS G12V/HLA-A*11:01 and KRAS WT/HLA-A*11:01. Binding kinetics were determined by fitting to a 1:1 Langmuir model. ( D ) Dose-dependent T-cell activation by scDb2. C1R-A11 cells pulsed with KRAS G12V or WT peptides were co-cultured with CD8 + T cells in the presence of increasing concentrations (0, 0.1, 0.3, 1 and 10 nM) of scDb2. IFN-γ secretion was quantified by ELISpot assay. Data represent mean±SD of three technical replicates. The EC 50 for IFN-γ secretion was 0.59 nM for C1R-A11 cells. p=0.0023 at 0.3 nM and p<0.0001 at 1 and 10 nM versus KRAS WT (Tukey’s test). ( E ) KRAS peptide-specific T-cell activation by scDb2. C1R-A11 cells were pulsed with different KRAS G12 variant peptides (WT, G12A, G12C, G12D, G12R and G12S), then co-cultured with CD8 + T cells in the presence of 1 nM KRAS G12V/A11:01-specific scDb2. IFN-γ secretion was measured by ELISpot. Data represent mean±SD of three technical replicates. p<0.0001 versus no peptide (Dunnett’s test). ( F ) HLA-restricted T-cell activation by scDb2. C1R cells expressing a panel of prevalent HLA-A alleles (HLA-A*02:01, HLA-A*02:06, HLA-A*03:01, HLA-A*11:01, HLA-A*24:02, HLA-A*31:01 and HLA-A*33:03) were pulsed with KRAS G12V peptide and co-cultured with CD8 + T cells in the presence of 1 nM KRAS G12V/A11:01-specific scDb2. Data represent mean±SD of three technical replicates. p<0.0001 for HLA-A*11:01 and HLA-A*03:01 versus the no-peptide control; no significant differences were observed for the other HLA alleles versus the no-peptide control (Dunnett’s test). BsAb, bispecific antibody; EC 50 , half-maximal effective concentrations; ELISpot, enzyme-linked immunosorbent spot; HLA, human leukocyte antigen; IFN, interferon; scDb, single-chain diabody; WT, wild-type.

Article Snippet: Sections were then incubated with primary antibodies: anti-CD8 mouse monoclonal antibody (Proteintech, Rosemont, Illinois, USA) and anti-Ki67 rabbit monoclonal antibody (Cell Signaling Technology, Danvers, Massachusetts, USA).

Techniques: Binding Assay, Purification, Enzyme-linked Immunosorbent Assay, Recombinant, Incubation, SPR Assay, Analysis, Activation Assay, Cell Culture, Enzyme-linked Immunospot, Variant Assay, Expressing, Control, ELISpot Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Engineering T cell-engaging bispecific antibodies targeting KRAS G12V neoantigen across HLA-A*11:01 and HLA-A*03:01

doi: 10.1136/jitc-2026-016411

Figure Lengend Snippet: Assessment of KRAS G12V/A*11:01-specific scDb2 specificity and potential cross-reactivity. ( A ) Cross-reactivity of KRAS G12V variant peptides. C1R-A11 cells were pulsed with the parental KRAS G12V peptide or amino acid-substituted variants, and co-cultured with CD8 + T cells in the presence of 1 nM KRAS G12V/A*11:01-specific scDb2. IFN-γ release was measured by ELISpot assay, and the mean of duplicate wells was used to generate the heatmap. Black boxes indicate the amino acids present in the parental G12V peptide. ( B ) Sequence specificity of KRAS G12V/A*11:01 scDb2 visualized as a Seq2Logo plot. Values were calculated by normalizing IFN-γ signals to a constant and plotting using the R package Seq2Logo. ( C ) AlphaFold-predicted structure of the KRAS G12V-scFv2 in complex with KRAS G12V/HLA-A*11:01. HLA-A*11:01, scFv and β2 microglobulin (β2M) are colored in gray, wheat and cyan, respectively. The KRAS G12V peptide is shown in light red between helices α1 and α2 of the HLA. ( D ) Bird’s-eye view of surface representation of the HLA-A*11:01 shown in gray, KRAS G12V peptide shown in red, and the contacting residues colored according to complementarity-determining regions (CDRs) of the scFv. The CDRs are labeled and colored as: H3 (pink), H2 (purple), H1 (sky blue), L3 (green), L2 (limon), L1 (dark green). ELISpot, enzyme-linked immunosorbent spot; HLA, human leukocyte antigen; IFN, interferon; scDb, single-chain diabody; scFv, single-chain variable fragments; WT, wild-type.

Article Snippet: Sections were then incubated with primary antibodies: anti-CD8 mouse monoclonal antibody (Proteintech, Rosemont, Illinois, USA) and anti-Ki67 rabbit monoclonal antibody (Cell Signaling Technology, Danvers, Massachusetts, USA).

Techniques: Variant Assay, Cell Culture, Enzyme-linked Immunospot, Sequencing, Labeling, ELISpot Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Engineering T cell-engaging bispecific antibodies targeting KRAS G12V neoantigen across HLA-A*11:01 and HLA-A*03:01

doi: 10.1136/jitc-2026-016411

Figure Lengend Snippet: In vivo antitumor efficacy of KRAS G12V-specific scDb2. ( A ) NSG mice were engrafted through s.c. injection with 5×10 6 SNG-M cells on day −7. On day 0, mice were randomized based on tumor sizes, and 2×10 7 human PBMCs or PBS as a vehicle control, and KRAS G12V-specific scDb2 or isotype control scDb (10 µg/mouse) were injected i.p. scDbs were subsequently injected two times per week. ( B ) Tumor progression was monitored by measuring tumor size. Data represent mean±SEM (n=8 mice/group). *p=0.043 PBMC+KRAS scDb2 versus PBMC+isotype scDb at the last time point (Tukey’s test). ( C ) Tumor weight of each mouse at the end of experiment was measured. Boxes show the median and IQR with individual mice plotted (n=8 mice/group). p values were calculated by Tukey’s test. ( D ) Representative H&E and multiplex immunohistochemical staining for CD8 and Ki67. Scale bars, 50 µm. ( E ) Proportion of CD8 + T cells and Ki67 + proliferating CD8 + T cells in tumors from each group. Data represent mean±SD (n=8 mice/group). p values were calculated by Tukey’s test. Scale bars, 10 µm. ( F ) Body weights of the mice were serially monitored in the established SNG-M model. Data represent mean±SEM (n=8 mice/group). (Tukey’s test, p=0.88 at the last time point). ( G ) Representative H&E staining of kidney and liver tissues from mice across treatment groups. Scale bars, 50 µm. i.p., intraperitoneally; NS, not significant; PBMC, peripheral blood mononuclear cells; PBS, phosphate buffered saline; s.c., subcutaneous; scDb, single-chain diabody.

Article Snippet: Sections were then incubated with primary antibodies: anti-CD8 mouse monoclonal antibody (Proteintech, Rosemont, Illinois, USA) and anti-Ki67 rabbit monoclonal antibody (Cell Signaling Technology, Danvers, Massachusetts, USA).

Techniques: In Vivo, Injection, Control, Multiplex Assay, Immunohistochemical staining, Staining, Saline