Journal: Molecular Cancer Therapeutics

Article Title: TPP-45142—an Anti-HER2 T-cell Engager—Designed for Selective HER2-Low Cancer Immunotherapy

doi: 10.1158/1535-7163.MCT-25-0654

Figure Lengend Snippet: TPP-45142 is a bispecific molecule that binds with a novel epitope of HER2. A, Schematic representation of TPP-45142. Green, two HER2-binding NANOBODY domains; orange, anti-TCRαβ NANOBODY domain; and gray, Fc domain with effectorless function. B, Cryo-EM structure of the complex HER2–29E09–Fab was obtained at 2.78 Å resolution. Left, colored electron density map. Right, full model. C, Cryo-EM structure of the 27A05–HER2–47D05–Fab complex was obtained at 2.66 Å resolution. Left, colored electron density map. Center, full model. Right, 27A05–HER2 interface. D, Structural superposition showing the relative location of pertuzumab and trastuzumab (based on PDB 6OGE) versus 27A05 and 29E09 as observed using cryo-EM. E, Structural superposition of 29E09 and 27A05. [ A, Created in BioRender. Vintem, A.P. (2026) https://BioRender.com/lk4spzo .]

Article Snippet: Human, cyno, and mouse HER2 Fc proteins were immobilized on a ProteOn GLC sensor chip (BioRad Laboratories, Inc. cat. #176-5011; 20 μg/mL, 10 mmol/L acetate pH 4.0, 120 seconds, 30 μL/minute).

Techniques: Binding Assay, Cryo-EM Sample Prep

Journal: Molecular Cancer Therapeutics

Article Title: TPP-45142—an Anti-HER2 T-cell Engager—Designed for Selective HER2-Low Cancer Immunotherapy

doi: 10.1158/1535-7163.MCT-25-0654

Figure Lengend Snippet: Cytotoxicity of TPP-45142 and its mechanism of action toward HER2-low breast cancer cell lines. A–D, TDCC of TPP-45142 for three T-cell donors compared with that of non-HER2 negative control (TPP-45161); co-cultures of human T cells with HCC1954, ZR-75-1, BT-20, or BT-549 cells were used at an E:T ratio of 5:1. E, TDCC of TPP-45142 for three T-cell donors compared with that of TPP-45161 in a co-culture of human T cells with BT20 3D spheroids at an E:T ratio of 1:5. F, T-cell activation induced by TPP-45142 as measured by expression of CD25 and CD69 expression on both CD4 + and CD8 + T cells as per FC analysis of ZR-75-1 and BT20 cells. G, Production of IFN-γ, IL2, IL6, IL8, IL10, and TNF-α cytokines in the culture supernatants obtained in the T-cell activation assay was measured using electrochemiluminescence assays.

Article Snippet: Human, cyno, and mouse HER2 Fc proteins were immobilized on a ProteOn GLC sensor chip (BioRad Laboratories, Inc. cat. #176-5011; 20 μg/mL, 10 mmol/L acetate pH 4.0, 120 seconds, 30 μL/minute).

Techniques: Negative Control, Co-Culture Assay, Activation Assay, Expressing, Electrochemiluminescence

Journal: Molecular Cancer Therapeutics

Article Title: TPP-45142—an Anti-HER2 T-cell Engager—Designed for Selective HER2-Low Cancer Immunotherapy

doi: 10.1158/1535-7163.MCT-25-0654

Figure Lengend Snippet: PK profiles and antitumor efficacy of TPP-45142 in the ZR-75-1 HER2-low breast cancer mouse model. A, TPP-45142 PK behavior in the ZR-75-1 xenograft model. Human T cells were administered to female NGS mice bearing intramammary ZR-75-1 tumors, and they were treated once with 89 Zr-TPP-45142 or the non-HER2 negative control 89 Zr-TPP-45161 ( n = 3). Microsamples (5 µL/time point) of blood were collected, and radioactivity was measured extemporaneously using a gamma counter (time: after radiolabeled-compound injection). B, Tumor accumulation of 89 Zr-TPP-45142 or non-HER2 negative control 89 Zr-TPP-45161 as measured by PET/CT imaging ( n = 3; time: after radiolabeled-compound injection). C, Antitumor activity of TPP-45142 in the ZR-75-1 xenograft model. Human T cells (10 × 10 6 ) were administered to female NSG mice bearing ZR-75-1 tumors, and they were treated on days 22 and 29 with TPP-45142 (500, 100, 50, and 10 μg/kg) and non-HER2 negative control TPP-45161 (500 μg/kg; n = 10 per group). ID, injected dose; MAD, median absolute deviation.

Article Snippet: Human, cyno, and mouse HER2 Fc proteins were immobilized on a ProteOn GLC sensor chip (BioRad Laboratories, Inc. cat. #176-5011; 20 μg/mL, 10 mmol/L acetate pH 4.0, 120 seconds, 30 μL/minute).

Techniques: Negative Control, Radioactivity, Injection, Positron Emission Tomography-Computed Tomography, Imaging, Activity Assay

Journal: Molecular Cancer Therapeutics

Article Title: TPP-45142—an Anti-HER2 T-cell Engager—Designed for Selective HER2-Low Cancer Immunotherapy

doi: 10.1158/1535-7163.MCT-25-0654

Figure Lengend Snippet: Safety profile of TPP-45142. A, T cell–mediated lysis of human cardiomyocytes was measured by impedance using xCELLigence. For donor 3, nine doses were tested in the range of 3 × 10 −11 to 5 × 10 −7 mol/L, whereas for donors 1 and 2, only the highest three doses were tested. B and C, HER2 distribution on the cell surface of BT-549 and HCM, respectively, via immunofluorescence. Arrowheads point to large HER2-enriched areas. D, Comparison of HER2 distribution between BT-549 ( B ) and HCM ( C ) cells via frequency distribution analysis of objects sorted by area. E, Heatmap of cytokine concentration as measured using the Luminex multiplex array after MIMIC CRA.

Article Snippet: Human, cyno, and mouse HER2 Fc proteins were immobilized on a ProteOn GLC sensor chip (BioRad Laboratories, Inc. cat. #176-5011; 20 μg/mL, 10 mmol/L acetate pH 4.0, 120 seconds, 30 μL/minute).

Techniques: Lysis, Immunofluorescence, Comparison, Concentration Assay, Luminex, Multiplex Assay

Journal: Cancer Research

Article Title: A Double-Negative Prostate Cancer Subtype Is Vulnerable to SWI/SNF-Targeting Degrader Molecules

doi: 10.1158/0008-5472.CAN-25-2928

Figure Lengend Snippet: TCF7L2 is a dependency in CRPC-WNT. A, Growth data measured by live-cell imaging (IncuCyte S3) upon transfection of indicated siRNA or plasmid (EV or rescue). Immunoblot of indicated proteins at 72 hours after transfection. GAPDH served as loading control. Data are presented as mean values ± SEM and analyzed using two-way ANOVA. ***, P < 0.001. Data are representative of n = 2 independent experiments. B, Spheroid formation measured by live-cell imaging (IncuCyte SX5) upon transfection of indicated siRNA. Data are presented as mean values ± SEM and analyzed using two-way ANOVA. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Data are representative of n = 2 independent experiments. C, IHC and staining intensity of TCF7L2 on indicated organoids upon treatment with 1 µmol/L A858 or 1 µmol/L A947 for 24 hours. Violin plot from TCF7L2 staining intensity analyzed using two-way ANOVA. ****, P < 0.0001. Scale bars, 100 µm (MSK-PCa16) and 50 µm (WCM1078). OD, optical density. D, TOPFlash TCF/LEF reporter assay measured after 48 hours upon treatment with indicated drugs in WCM1078 sublines. FOPFlash served as a negative reporter control. Data are presented as mean values ± SEM after normalization to internal Renilla control and analyzed using a paired Student t test. ***, P < 0.001; ****, P < 0.0001. Data are representative of n = 2 independent experiments.

Article Snippet: Chromatin was prepared from two biological replicates of WCM1078 treated with A858 or A947 (1 μmol/L) for 4 hours, and chromatin immunoprecipitation sequencing (ChIP-seq) assays were then performed using an antibody against TCF7L2 (Cell Signaling Technology, cat #2569, RRID: AB_2199816).

Techniques: Live Cell Imaging, Transfection, Plasmid Preparation, Western Blot, Control, Staining, Reporter Assay

Journal: Cancer Research

Article Title: A Double-Negative Prostate Cancer Subtype Is Vulnerable to SWI/SNF-Targeting Degrader Molecules

doi: 10.1158/0008-5472.CAN-25-2928

Figure Lengend Snippet: TCF7L2 regulates proproliferative signatures in CRPC-WNT. A, ChIP-seq read density tornado plots from WCM1078 organoids treated with 1 µmol/L A858 or 1 µmol/L A947 for 4 hours ( n = 2 biological replicates). B, Venn diagram indicating A947 treatment–lost regions from ChIP-seq, ATAC-seq, and RNA-seq data in WCM1078. GSEA was performed from 350 overlapping genes. C, Immunoblot of indicated proteins at indicated time upon treatment with 1 µmol/L A947. GAPDH served as loading control. Data are representative of n = 2 independent experiments. D, Dose–response curves with indicated drugs after measurement of proliferation with CellTiter-Glo 2.0 after 7-day treatment ( n = 2 independent experiments).

Article Snippet: Chromatin was prepared from two biological replicates of WCM1078 treated with A858 or A947 (1 μmol/L) for 4 hours, and chromatin immunoprecipitation sequencing (ChIP-seq) assays were then performed using an antibody against TCF7L2 (Cell Signaling Technology, cat #2569, RRID: AB_2199816).

Techniques: ChIP-sequencing, RNA Sequencing, Western Blot, Control

Journal: Redox Biology

Article Title: NMRK2–YAP–NADK axis preserves redox protection against myocardial ischemia/reperfusion injury

doi: 10.1016/j.redox.2026.104100

Figure Lengend Snippet: YAP directly binds to the NADK promoter and activates its transcription. (A) Schematic illustration of the human NADK promoter region (−2000 to −0 bp relative to the transcription start site) showing putative YAP-binding sites and the design of a series of truncated NADK promoter luciferase reporter constructs. (B) ChIP-qPCR analysis demonstrating significant enrichment of the NADK promoter region in anti-YAP immunoprecipitates compared with IgG controls ( n = 3). (C) Luciferase reporter assays showing that YAP overexpression significantly enhances transcriptional activity driven by the −2000/0 and −1500/0 NADK promoter fragments, whereas further truncation to −1000/0 or −500/0 markedly attenuates YAP-induced activation, identifying the YAP-responsive region within the −1500 to −1000 bp interval ( n = 3). Data are presented as mean ± SEM. Unpaired Student's t -test or one-way ANOVA followed by Tukey's post hoc test for comparisons. *** P < 0.001. ChIP, Chromatin immunoprecipitation; IgG, immunoglobulin G; LUC, luciferase; Rluc, Renilla luciferase; NC, negative control.

Article Snippet: ChIP assays were performed using a commercial ChIP kit (CST 9005S) to assess the binding of YAP to the NADK promoter.

Techniques: Binding Assay, Luciferase, Construct, ChIP-qPCR, Over Expression, Activity Assay, Activation Assay, Chromatin Immunoprecipitation, Negative Control