Journal: Bioengineering & Translational Medicine

Article Title: Combined HER2 ‐targeted mRNA ‐ LNP vaccine and radiotherapy suppress Lewis lung carcinoma growth in mice: Efficacy and mechanistic insights

doi: 10.1002/btm2.70160

Figure Lengend Snippet: (a) Plasmid map of the HER2 extracellular domain (ECD). (b) Flowchart of HER2‐mRNA‐LNP preparation. (c) Agarose gel electrophoresis of HER2 ECD mRNA (double‐site digested product). (d) Agarose gel electrophoresis of HER2 ECD total mRNA (single‐site digested product). (e) Particle size and zeta potential of blank LNPs and HER2‐mRNA‐LNPs. (f) Representative cryo‐TEM micrographs of HER2‐mRNA‐LNPs at magnifications of 28,000×, 45,000×, and 92,000×. (g) Encapsulation efficiency of HER2‐mRNA‐LNPs. (h) Western blot analysis of HER2 ECD protein expression, with relative grayscale values quantified and statistically analyzed by Student's t ‐test ( n = 3 independent experiments). (i) Serum anti‐HER2 antibody titers in BALB/c and C57BL/6 mice immunized with LNP or HER2‐mRNA‐LNP after three doses. Data are presented as mean ± SEM. n = 3 mice per group. Statistical significance was determined by two‐way repeated‐measures ANOVA followed by Tukey's multiple comparisons test. **** p < 0.0001.

Article Snippet: Human HER2 ECD (Sino Biological; 0.25 mg/mL) was coated onto an ELISA plate overnight at 20 ng/well.

Techniques: Plasmid Preparation, Agarose Gel Electrophoresis, Zeta Potential Analyzer, Cryo-TEM, Encapsulation, Western Blot, Analysis, Expressing

Journal: Bioengineering & Translational Medicine

Article Title: Combined HER2 ‐targeted mRNA ‐ LNP vaccine and radiotherapy suppress Lewis lung carcinoma growth in mice: Efficacy and mechanistic insights

doi: 10.1002/btm2.70160

Figure Lengend Snippet: (a) Gross images of liver, lung, heart, and spleen from mice treated with LNP or HER2‐mRNA‐LNP. (b) Body weight and organ weights (liver, lung, heart, and spleen) of mice in the two groups. (c) H&E staining of liver, lung, heart and spleen tissue sections (magnification: 200× and 400×). (d) Peripheral blood routine analysis: white blood cell (WBC), red blood cell (RBC), and platelet (PLT) counts. (e) Serum liver function parameters: alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), γ‐glutamyl transpeptidase (γ‐GT), albumin (ALB), direct bilirubin (DBIL), total bilirubin (TBIL), and total bile acid (TBA). (f) Serum renal function parameters: Urea (UREA), creatinine (CREA), and uric acid (UA). (g) Serum myocardial enzyme profile: creatine kinase (CK (U/L)), creatine kinase‐MB (CK‐MB), lactate dehydrogenase (LDH), and lactate dehydrogenase isoenzyme 1 (LDH1). All data are presented as mean ± SEM, with n = 5 mice per group. Statistical analysis was performed using unpaired two‐tailed Student's t ‐test. ** p < 0.01; ns, not significant ( p > 0.05).

Article Snippet: Human HER2 ECD (Sino Biological; 0.25 mg/mL) was coated onto an ELISA plate overnight at 20 ng/well.

Techniques: Staining, Tissue, Analysis, Two Tailed Test

Journal: Bioengineering & Translational Medicine

Article Title: Combined HER2 ‐targeted mRNA ‐ LNP vaccine and radiotherapy suppress Lewis lung carcinoma growth in mice: Efficacy and mechanistic insights

doi: 10.1002/btm2.70160

Figure Lengend Snippet: (a) HER2 mRNA expression in LLC and HER2‐LLC cells detected by Q‐PCR; HER2 protein expression in LLC, HER2‐LLC, SKBR3, and A549 cells analyzed by Western blot. (b) Anti‐HER2 antibody titers in mice after three doses of HER2‐mRNA‐LNP. (c) Tumor growth curve of HER2‐LLC tumor‐bearing mice in each group. (d) Treatment schedule for each group of mice. (e) Representative tumor images from each group. (f) Tumor volume and weight in each group. (g) CCK‐8 assay‐based inhibition rates of HER2‐LLC cells by splenic lymphocytes from tumor‐bearing mice. (h) Correlation analysis of tumor volume and antibody titer in HER2‐mRNA‐LNP group mice. (i) Quantitative immunohistochemical analysis of HER2 and Ki67 expression in tumor tissues from each group of mice. Three fields of view were selected per mouse, yielding 15 data points per group. (j) Representative immunohistochemical images of HER2 and Ki67 expression in mouse tumor tissues (magnification: 200× and 400×). (k) Kinetics of antibody titers over time in the PBS group, 1‐dose vaccine group, and 3‐dose vaccine group. All data are presented as mean ± SEM, with n = 5 mice per group. One‐way ANOVA with Tukey's test or Kruskal–Wallis test with Dunn's test was used for multiple group comparisons, as appropriate; statistical analysis was performed using two‐way repeated‐measures ANOVA followed by Tukey's multiple comparisons test for antibody titers, tumor growth curves and CCK‐8 inhibition rates; Pearson correlation analysis for correlation analyses ( R = −0.8889, p = 0.0437). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant ( p > 0.05).

Article Snippet: Human HER2 ECD (Sino Biological; 0.25 mg/mL) was coated onto an ELISA plate overnight at 20 ng/well.

Techniques: Expressing, Western Blot, CCK-8 Assay, Inhibition, Analysis, Titer Analysis, Immunohistochemical staining

Journal: Bioengineering & Translational Medicine

Article Title: Combined HER2 ‐targeted mRNA ‐ LNP vaccine and radiotherapy suppress Lewis lung carcinoma growth in mice: Efficacy and mechanistic insights

doi: 10.1002/btm2.70160

Figure Lengend Snippet: Results of two‐way repeated‐measures ANOVA for serum HER2 antibody titer levels.

Article Snippet: Human HER2 ECD (Sino Biological; 0.25 mg/mL) was coated onto an ELISA plate overnight at 20 ng/well.

Techniques: Titer Analysis

Journal: Bioengineering & Translational Medicine

Article Title: Combined HER2 ‐targeted mRNA ‐ LNP vaccine and radiotherapy suppress Lewis lung carcinoma growth in mice: Efficacy and mechanistic insights

doi: 10.1002/btm2.70160

Figure Lengend Snippet: (a) Experimental timeline for each group. (b) Tumor growth curves of HER2‐LLC tumor‐bearing mice. (c) Representative tumor images. (d) Quantification of tumor volume and weight ( n = 5). (e) CCK‐8 assay‐based cytotoxicity inhibition rate at 48 h ( n = 3). (f) Representative immunohistochemical images of HER2 and Ki67 expression in mouse tumor tissues. (g) Quantitative analysis of HER2 and Ki67 immunohistochemical staining across groups (Three random fields per mouse were selected for analysis. n = 5). All data are presented as mean ± SEM. Two‐way repeated‐measures ANOVA with Tukey's test was used for tumor growth curves ( n = 5) (b); one‐way ANOVA with Tukey's test was used for multiple group comparisons (d,e,g).

Article Snippet: Human HER2 ECD (Sino Biological; 0.25 mg/mL) was coated onto an ELISA plate overnight at 20 ng/well.

Techniques: CCK-8 Assay, Inhibition, Immunohistochemical staining, Expressing, Analysis, Staining

Journal: Scientific Reports

Article Title: Machine learning-driven drug repurposing for HER2-positive breast cancer

doi: 10.1038/s41598-026-45361-z

Figure Lengend Snippet: Mechanism of HER2 in breast cancer.

Article Snippet: Fig. 7 Molecular Dynamics of Timolol maleate complex with wild-type HER2 protein for 1000ns simulation ( a ) RMSD, ( b ) RMSF, ( c ) Protein-Ligand Contacts, and ( d ) Ligand-Protein Contacts.

Techniques:

Journal: Scientific Reports

Article Title: Machine learning-driven drug repurposing for HER2-positive breast cancer

doi: 10.1038/s41598-026-45361-z

Figure Lengend Snippet: 2D and 3D amino acid interactions of the top 5 selected ligands complexed with the wild HER2 protein (7PCD).

Article Snippet: Fig. 7 Molecular Dynamics of Timolol maleate complex with wild-type HER2 protein for 1000ns simulation ( a ) RMSD, ( b ) RMSF, ( c ) Protein-Ligand Contacts, and ( d ) Ligand-Protein Contacts.

Techniques:

Journal: Scientific Reports

Article Title: Machine learning-driven drug repurposing for HER2-positive breast cancer

doi: 10.1038/s41598-026-45361-z

Figure Lengend Snippet: 2D and 3D amino acid interactions of the top 5 selected ligands complexed with the mutated HER2 protein (8U8X).

Article Snippet: Fig. 7 Molecular Dynamics of Timolol maleate complex with wild-type HER2 protein for 1000ns simulation ( a ) RMSD, ( b ) RMSF, ( c ) Protein-Ligand Contacts, and ( d ) Ligand-Protein Contacts.

Techniques:

Journal: Scientific Reports

Article Title: Machine learning-driven drug repurposing for HER2-positive breast cancer

doi: 10.1038/s41598-026-45361-z

Figure Lengend Snippet: Molecular Dynamics of Timolol maleate complex with wild-type HER2 protein for 1000ns simulation ( a ) RMSD, ( b ) RMSF, ( c ) Protein-Ligand Contacts, and ( d ) Ligand-Protein Contacts.

Article Snippet: Fig. 7 Molecular Dynamics of Timolol maleate complex with wild-type HER2 protein for 1000ns simulation ( a ) RMSD, ( b ) RMSF, ( c ) Protein-Ligand Contacts, and ( d ) Ligand-Protein Contacts.

Techniques:

Journal: Scientific Reports

Article Title: Machine learning-driven drug repurposing for HER2-positive breast cancer

doi: 10.1038/s41598-026-45361-z

Figure Lengend Snippet: Molecular Dynamics of Timolol maleate complex with mutated type HER2 protein for 1000ns simulation ( a ) RMSD, ( b ) RMSF, ( c ) Protein-Ligand Contacts & ( d ) Ligand-Protein Contacts.

Article Snippet: Fig. 7 Molecular Dynamics of Timolol maleate complex with wild-type HER2 protein for 1000ns simulation ( a ) RMSD, ( b ) RMSF, ( c ) Protein-Ligand Contacts, and ( d ) Ligand-Protein Contacts.

Techniques:

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: Frontiers in Immunology

Article Title: Robust CD8 + T cell responses induced by an mRNA-LNP vaccine encoding rat HER2 extracellular domain confer prophylactic tumor protection

doi: 10.3389/fimmu.2026.1737558

Figure Lengend Snippet: mRNA-LNP vaccine development via integrated mechanism, preparation, and characterization. (A) Schematic of the mRNA-LNP vaccine mechanism. Intramuscularly injected mRNA-LNPs are delivered to DCs. The encoded antigen is expressed and processed through two major pathways: the immunoproteasome/MHC class I pathway activates CD8 + T cells, while the endosomal/MHC class II pathway activates CD4 + T cells. Activated CD4 + T cells provide help to B cells for antibody production, orchestrating a coordinated adaptive immune response. (B) Schematic microfluidic process of mRNA-LNPs preparation. (C) Plasmid map of rat HER2 ECD and rat HER2 ECD-IFNγ. (D) Agarose gel electrophoresis of plasmid and linearized plasmid DNA. (E) Agarose gel electrophoresis of transcribed mRNA and total mRNA for rat HER2 ECD and rat HER2 ECD-IFNγ. M, 1kb ladder; 1, Plasmids with rat HER2 ECD sequence and 2, rat HER2 ECD-IFNγ sequence; 3, Linearized plasmid DNA with rat HER2 ECD sequence and 4, rat HER2 ECD-IFNγ sequence; 5, Agarose gel electrophoresis images of transcribed mRNA and 6, total mRNA of rat HER2 ECD; 7, Agarose gel electrophoresis images of transcribed mRNA and 8, total mRNA of rat HER2 ECD-IFNγ.

Article Snippet: Splenocytes (2 × 10 5 cells/well) were stimulated in complete RPMI 1640 medium with or without 2 μg/mL rat HER2 protein (#80079-R08H, Sino Biological) for 12 hours at 37 °C.

Techniques: Bioprocessing, Injection, Plasmid Preparation, Agarose Gel Electrophoresis, Sequencing

Journal: Frontiers in Immunology

Article Title: Robust CD8 + T cell responses induced by an mRNA-LNP vaccine encoding rat HER2 extracellular domain confer prophylactic tumor protection

doi: 10.3389/fimmu.2026.1737558

Figure Lengend Snippet: Therapeutic efficacy of HER2 ECD mRNA-LNP vaccines in 4T1-HER2 tumor-bearing mice. (A) Treatment schedules and dosing intervals of the 4T1-HER2 tumor model. (B) Serum anti-HER2 antibody kinetics after 5 μg or 10 μg rHER2 ECD mRNA-LNP vaccination. (C) Changes in serum HER2 antibody concentration over time after administration after rHER2 ECD mRNA-LNP or rHER2 ECD-IFNγ mRNA-LNP vaccination. (D) Growth curve of 4T1-HER2 tumors. (E) Representative images of tumor volume. N = 3. Data are presented as mean ± standard error of the mean (SEM); two-sided t-test. (* P < 0.05 and *** P < 0.001).

Article Snippet: Splenocytes (2 × 10 5 cells/well) were stimulated in complete RPMI 1640 medium with or without 2 μg/mL rat HER2 protein (#80079-R08H, Sino Biological) for 12 hours at 37 °C.

Techniques: Drug discovery, Vaccines, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Robust CD8 + T cell responses induced by an mRNA-LNP vaccine encoding rat HER2 extracellular domain confer prophylactic tumor protection

doi: 10.3389/fimmu.2026.1737558

Figure Lengend Snippet: Safety profiles of HER2 ECD mRNA-LNP vaccines in 4T1-HER2 tumor-bearing mice. (A) Body, heart, liver and lung weights of mice. (B) Hepatic function, renal function and cardiac function on day 68. N = 3. Data are presented as mean ± SEM; two-sided t-test. No significant intergroup differences were observed.

Article Snippet: Splenocytes (2 × 10 5 cells/well) were stimulated in complete RPMI 1640 medium with or without 2 μg/mL rat HER2 protein (#80079-R08H, Sino Biological) for 12 hours at 37 °C.

Techniques: Vaccines

Journal: Frontiers in Immunology

Article Title: Robust CD8 + T cell responses induced by an mRNA-LNP vaccine encoding rat HER2 extracellular domain confer prophylactic tumor protection

doi: 10.3389/fimmu.2026.1737558

Figure Lengend Snippet: Memory T cell subsets and antitumor activity. (A) Representative flow cytometry plots of T CM cells (CD44 + CD62L + ), T EM cells (CD44 + ) and naive cells (CD44 - CD62L + ) within CD4 + and (B) CD8 + T cells. (C) Rate of inhibition of 4T1-HER2 cells by vaccinated mouse splenic lymphocytes. (D) Concentration of IFNγ antibody in serum after administration. N = 3. Data are presented as mean ± SEM; two-sided t-test. (* P < 0.05, ** P < 0.01, and *** P < 0.001).

Article Snippet: Splenocytes (2 × 10 5 cells/well) were stimulated in complete RPMI 1640 medium with or without 2 μg/mL rat HER2 protein (#80079-R08H, Sino Biological) for 12 hours at 37 °C.

Techniques: Activity Assay, Flow Cytometry, Inhibition, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Robust CD8 + T cell responses induced by an mRNA-LNP vaccine encoding rat HER2 extracellular domain confer prophylactic tumor protection

doi: 10.3389/fimmu.2026.1737558

Figure Lengend Snippet: Antitumor efficacy of combination therapy. (A) Treatment schedules and dosing intervals of the 4T1-HER2 tumor model. (B) Changes in serum HER2 antibody concentration over time after administration after rHER2 ECD mRNA-LNP vaccination, with and without anti-PD-1. (C) Growth curve of 4T1-HER2 tumors. (D) Representative images of tumor tissue. (E) Tumor volume. N = 6;Data are presented as mean ± SEM. Two-sided t-test. (* P < 0.05 and *** P < 0.001).

Article Snippet: Splenocytes (2 × 10 5 cells/well) were stimulated in complete RPMI 1640 medium with or without 2 μg/mL rat HER2 protein (#80079-R08H, Sino Biological) for 12 hours at 37 °C.

Techniques: Concentration Assay