Journal: Nature Communications

Article Title: The HER2-directed antibody-drug conjugate DHES0815A in advanced and/or metastatic breast cancer: preclinical characterization and phase 1 trial results

doi: 10.1038/s41467-023-44533-z

Figure Lengend Snippet: a Structures of PBD, PBD-monoamine and PBD-monoamide, and summary table for DNA binding and drug activity (free drugs and ADCs in vitro and in vivo). In the left panel, circles denote reactive imine moiety and changes thereof to generate monoamine and monoamide derivatives. For the summary table: large cell line screens were performed to assess potency of the PBD dimer, PBD-monoamine and -monoamide. The PBD dimer was tested in 7 different cell line screens; the PBD-monoamide in 3 screens; and the PBD-monoamine in 2 screens ( n = 3 wells per treatment group for all). The number of cell lines tested per screen ranged from 50-643 for the PBD; 73-146 for the PBD-monoamide and 72-147 for the PBD-monoamine. Data shown are the pooled mean IC 50 values ± standard error from the respective screens. ADC IC 50 was determined in SK-BR-3 cells (data are mean IC 50 for 3 independent experiments, n = 4 wells per treatment group for each; see Fig. 1c for graph). DNA alkylation percent is derived from data shown in part 1b. For determining MED, multiple xenograft models were used, with n = 5 to n = 10 mice per group. MED = minimum efficacious dose, defined as a single injection dose that results in tumor stasis at day 21 in mouse xenograft models. b DNA binding of PBD dimer, PBD-monoamine and PBD-monoamide as assessed by alkylation of double-stranded DNA oligonucleotides (each point represents oligonucleotide reactions, from 2 separate experiments). c Activity of DHES0815A compared to conjugated PBD dimer or PBD-monoamine (same antibody and linker) in SK-BR-3 cells in vitro and in MMTV-HER2 Fo5 model in vivo (PBD and PBD-monoamide conjugates only, n = 8 mice per group). d Structure of DHES0815A, comprised of 7C2 THIOMAB (MHES0448A), methyl-disulfide linker and PBD-monoamide. Source data are provided as a Source Data file.

Article Snippet: PBD-containing ADCs were conjugated to linker-drugs through engineered cysteines at light chain K149 using a nitropyridyl methyl-disulfide linker at Genentech.

Techniques: Binding Assay, Activity Assay, In Vitro, In Vivo, Derivative Assay, Injection

Journal: Nature Communications

Article Title: The HER2-directed antibody-drug conjugate DHES0815A in advanced and/or metastatic breast cancer: preclinical characterization and phase 1 trial results

doi: 10.1038/s41467-023-44533-z

Figure Lengend Snippet: a Kinetics of caspase 3 activation for DHES0815A compared to T-DM1, MHES0448A and trastuzumab ( n = 6 individual wells for each treatment group, the study was repeated 4 times). b Time-dependent PARP cleavage after treatment with antibodies (trastuzumab, MHES0488 A) or ADCs (T-DM1 or DHES0815A). c Induction of DNA damage markers (phospho-H2AX, phospho-/total p53 and phospho-CHK2) with free PBD-monoamide and DHES0815A. d Time-dependent induction of DNA damage and mitotic (phospho-histone H3) markers after treatment with unconjugated antibodies and ADCs. Western blot studies in ( b )–( d ), were performed 2 times with the same results. Source data are provided in Source Data file.

Article Snippet: PBD-containing ADCs were conjugated to linker-drugs through engineered cysteines at light chain K149 using a nitropyridyl methyl-disulfide linker at Genentech.

Techniques: Activation Assay, Western Blot

Journal: Materials Today Bio

Article Title: Ingenious designed a HER2-Specific macrophage biomimetic multifunctional nanoplatform for enhanced bio-photothermal synergistic therapy in HER2 positive breast cancer

doi: 10.1016/j.mtbio.2024.101095

Figure Lengend Snippet: Illustration of antiHER2 engineered macrophage biomimetic photothermal (AMBP) systems for photothermal/bio therapy of cancer. A) Schematic illustration of anti-HER2 expressed macrophages. The Fabrication of B) AMBP NPs and C) the mechanisms of AMBP NPs-mediated photothermal/bio therapy of cancer.

Article Snippet: However, with the combination of the photothermal effect and anti-HER2 induced biotherapeutics (G7), cell viability was reduced substantially to 25 %.

Techniques:

Journal: Materials Today Bio

Article Title: Ingenious designed a HER2-Specific macrophage biomimetic multifunctional nanoplatform for enhanced bio-photothermal synergistic therapy in HER2 positive breast cancer

doi: 10.1016/j.mtbio.2024.101095

Figure Lengend Snippet: Construction of anti-HER2 expressed macrophages. A) Schematic illustration of anti-HER2 expressed macrophages. B) The mRNA expression level of HER2-scFV-macrophages. C) CLSM results of HER2-scFV-GFP-macrophages with DiI to verify their transmembrane expression. HER-2-scFv-GFP were presented with green, macrophage was labeled with DiI (Red), and nuclei were stained with DAPI (Blue). D) The Structure of the anti-HER2 protein domain. E) The Binding areas determined from molecular docking of HER2 (red region) and anti-HER2 (gray region). F) The Binding results obtained by molecular dynamic simulation in this work. G) The Residue interaction between HER2 and anti-HER2 obtained by molecular dynamic simulation. H) Cross-correlation matrices of the coordinates' fluctuations for Cα atoms around their mean positions during the equilibrium simulation. Red color represents correlated motions and blue color represents anti-correlated motions. Results are presented as means ± SD. *** P < 0.001.

Article Snippet: However, with the combination of the photothermal effect and anti-HER2 induced biotherapeutics (G7), cell viability was reduced substantially to 25 %.

Techniques: Expressing, Labeling, Staining, Binding Assay, Residue

Journal: Materials Today Bio

Article Title: Ingenious designed a HER2-Specific macrophage biomimetic multifunctional nanoplatform for enhanced bio-photothermal synergistic therapy in HER2 positive breast cancer

doi: 10.1016/j.mtbio.2024.101095

Figure Lengend Snippet: Cellular uptake and immune escape of AMBP NPs. A) Cell Viability of AMBP NPs on NIH-3T3 cell lines. B) Hemolysis analysis of red blood cells after incubation with AMBP NPs at different concentrations. C) CLSM results of HER2 + -4T1 cells incubated with different format nanomaterials to verify their cellular uptake performance. NPs were labeled with Rh6G (Red) and nuclei were stained with DAPI (Blue). Flow cytometry assay of D) HER2 + -4T1 cells and E) SKOV3 cells incubated with different formats to verify their cellular uptake behavior. F) CLSM results and of macrophages incubated with different formats to demonstrate their immune escape ability of macrophages. NPs were labeled with Rh6G (Red) and nuclei were stained with DAPI (Blue). ZIF-8 enhanced the uptake of Rh6G by macrophages whereas the macrophage coating reduced the uptake of Rh6G@ZIF-8 by macrophages after each group co-culturing with macrophages, respectively. G) The Mean fluorescence intensity by flow cytometry analysis to show their immune escape behaviors. Results are presented as means ± SD. *** P < 0.001, n.s., not significant, P > 0.05.

Article Snippet: However, with the combination of the photothermal effect and anti-HER2 induced biotherapeutics (G7), cell viability was reduced substantially to 25 %.

Techniques: Incubation, Labeling, Staining, Flow Cytometry, Fluorescence

Journal: Materials Today Bio

Article Title: Ingenious designed a HER2-Specific macrophage biomimetic multifunctional nanoplatform for enhanced bio-photothermal synergistic therapy in HER2 positive breast cancer

doi: 10.1016/j.mtbio.2024.101095

Figure Lengend Snippet: In vitro Anti-tumor Effect of AMBP NPs. A) Cell viability of AMBP NPs on HER2 + -4T1 cell lines. B) Cell viability of AMBP NPs on SKOV3 cell lines. qRT-PCR assays of C) PI3K mRNA, D) AKT mRNA, E) p53 mRNA, F) Caspase 3 mRNA, G) Ki67 mRNA, and H) CCND-1 mRNA in HER2 + -4T1 cells with different treatments. I) Schematic illustration shows their bio-photothermal synergistic therapy mechanism. J) Cell viability of nanomaterials on HER2 + -4T1 cell lines to show their combined therapeutic effect. (G1: PBS, G2: Laser, G3: IR820, G4: ZIF-8@EM φ , G5: IR820@ZIF-8, G6: MBP NPs, G7: AMBP NPs). K) CLSM images of HER2 + -4T1 cells stained with calcein AM (green, live cells) and propidium iodide (red, dead cells) after different treatments. Scale bar = 50 μm. Data are expressed as the mean ± s.d. (* P < 0.05, ** P < 0.01, and *** P < 0.001).

Article Snippet: However, with the combination of the photothermal effect and anti-HER2 induced biotherapeutics (G7), cell viability was reduced substantially to 25 %.

Techniques: In Vitro, Quantitative RT-PCR, Staining

Journal: Materials Today Bio

Article Title: Ingenious designed a HER2-Specific macrophage biomimetic multifunctional nanoplatform for enhanced bio-photothermal synergistic therapy in HER2 positive breast cancer

doi: 10.1016/j.mtbio.2024.101095

Figure Lengend Snippet: Pharmacokinetics and biodistribution of AMBP NPs in vivo . A) Schematic illustration shows their protocols. B) Pharmacokinetic profiles of IR820@ZIF-8, MBP, and AMBP NPs in normal BALB/c mice (Ex: 710 nm, Em: 820 nm). C) In vivo fluorescence imaging of IR820@ZIF-8, MBP and AMBP NPs in HER2 + -4T1 tumor-bearing mice (Ex: 710 nm, Em: 820 nm). D) The Fluorescence intensity within the tumor region after different treatments. E) Ex vivo fluorescence images of major organs (heart, liver, spleen, lung, kidney) at 8 h post-injection. F) Biodistribution of IR820@ZIF-8, MBP and AMBP NPs in HER2 + -4T1 tumor-bearing mice at 8 h post-injection. G) Ex vivo fluorescence images of tumors under different treatments. H) Infrared thermal images of PBS, IR820@ZIF-8, MBP and AMBP NPs upon 808 nm laser (1W cm −2 ) irradiation for 5 min. I) The Temperature elevation curves of different treatments. Results are presented as means ± SD. *** P < 0.001, n.s., not significant, P > 0.05.

Article Snippet: However, with the combination of the photothermal effect and anti-HER2 induced biotherapeutics (G7), cell viability was reduced substantially to 25 %.

Techniques: Drug discovery, In Vivo, Fluorescence, Imaging, Ex Vivo, Injection, Irradiation

Journal: Materials Today Bio

Article Title: Ingenious designed a HER2-Specific macrophage biomimetic multifunctional nanoplatform for enhanced bio-photothermal synergistic therapy in HER2 positive breast cancer

doi: 10.1016/j.mtbio.2024.101095

Figure Lengend Snippet: Tumor therapy of AMBP NPs in vivo . A) Schematic illustration shows their protocols. B) Images of HER2 + -4T1 tumor-bearing mice under different treatments (i.e. PBS, control group, IR820@ZIF-8: phototherapeutic group, ZIF-8@EM φ : biotherapeutic group, MBP NPs, AMBP NPs, photothermal therapy and biotherapy). C) The Average tumor volume curves of mice after various treatments. D) Individual tumor growth curves of mice after various treatments. E) Images of HER2 + -4T1 tumor-bearing mice under different treatments. F) Tumor weight of HER2 + -4T1 tumor-bearing mice with different treatments. G) The Mice weight curves. qRT-PCR assays of H) PI3K mRNA, I) AKT mRNA in HER2 + -4T1 tumor-bearing mice. J) H&E staining of major mice organs. Data are expressed as the mean ± s.d. (** P < 0.01, and *** P < 0.001).

Article Snippet: However, with the combination of the photothermal effect and anti-HER2 induced biotherapeutics (G7), cell viability was reduced substantially to 25 %.

Techniques: In Vivo, Control, Quantitative RT-PCR, Staining