Journal: Cancer Medicine

Article Title: In Vivo Immunoscoring Technique to Monitor Anti‐ CD47 Immunotherapy Efficacy in Triple‐Negative Breast Cancer

doi: 10.1002/cam4.72232

Figure Lengend Snippet: Real‐time monitoring of tumor progression and anti‐CD47 therapeutic efficiency. (A) Female BALB/c mice aged 8–10 weeks were implanted with dorsal skinfold window chambers. 4T1_PB3R‐RFP TNBC cells were transplanted into the subcutaneous tissue, and tumors were monitored using intravital imaging. (B) Detailed visualization of angiogenesis and tumor nodule formation in TNBC skinfold models compared with the isotype control group. (C, D) Conventional whole‐body bioluminescence imaging of mice bearing 4T1_PB3R‐RFP‐luc tumors revealed considerably decreased signals following anti‐CD47 immunotherapy administered over 5 days, beginning on day 9 for 5 days ( n = 7, p < 0.0001, 10 mg/kg, intravenous injection). Data are presented as the mean ± standard deviation (SD).

Article Snippet: To evaluate the performance of the in vivo immunoscoring technique for the monitoring of anti‐CD47 immunotherapy efficacy, anti‐mouse CD47 monoclonal antibodies (specifically, anti‐CD47 mAbs, clone MIAP301, Rat IgG2a, κ, BioXcell, USA) and an IgG2a mAbs isotype control (clone 2A3, Rat IgG2a, κ, BioXcell, USA) were systemically and intravenously (IV) injected at a dosage of 10 mg/kg once a day for a duration of five consecutive days.

Techniques: Real-time Monitoring, Tissue, Imaging, Control, Injection, Standard Deviation

Journal: Cancer Medicine

Article Title: In Vivo Immunoscoring Technique to Monitor Anti‐ CD47 Immunotherapy Efficacy in Triple‐Negative Breast Cancer

doi: 10.1002/cam4.72232

Figure Lengend Snippet: In vivo and in vitro validation of TNBC targeting by Dextran‐AF680. (A–C) The signals emitted by Dextran‐AF680 at various time intervals following intravenous injections exhibited significant differences between 10 and 180 min and between 180 and 360 min. (D, E) Dextran‐AF680 can be administrated for TNBC imaging with photoacoustic tomography, and the optimal imaging time is 3 h after dextran injection, indicating its potential as an imaging probe for enhancing TNBC visualization ( n = 7, p = 0.024, p = 0.004). (F, G) Longitudinal conventional fluorescence imaging of the anti‐CD47 immunotherapeutic response, in comparison with the isotype control treatment, demonstrated a significant change in the average radiant efficiency of Dextran AF680 following 5 days of treatment ( n = 12, p = 0.0022). (H, I) An evident shift in the population of Dextran‐AF680+ and RFP+ cells was observed after anti‐CD47 mAb immunotherapies ( n = 10, p = 0.0079). Data are presented as the mean ± standard deviation (SD).

Article Snippet: To evaluate the performance of the in vivo immunoscoring technique for the monitoring of anti‐CD47 immunotherapy efficacy, anti‐mouse CD47 monoclonal antibodies (specifically, anti‐CD47 mAbs, clone MIAP301, Rat IgG2a, κ, BioXcell, USA) and an IgG2a mAbs isotype control (clone 2A3, Rat IgG2a, κ, BioXcell, USA) were systemically and intravenously (IV) injected at a dosage of 10 mg/kg once a day for a duration of five consecutive days.

Techniques: In Vivo, In Vitro, Biomarker Discovery, Imaging, Micro-PAT, Injection, Fluorescence, Comparison, Control, Standard Deviation

Journal: Cancer Medicine

Article Title: In Vivo Immunoscoring Technique to Monitor Anti‐ CD47 Immunotherapy Efficacy in Triple‐Negative Breast Cancer

doi: 10.1002/cam4.72232

Figure Lengend Snippet: Intravital imaging of M2 macrophage targeting by Dextran‐AF680. (A) Dynamic in vivo distribution of intravenously injected Dextran‐AF680 in representative blood vessels of a skinfold TNBC model. The Dextran‐AF680 signal peaked at 3 h and dissipated from the vessels within 24 h. (B) Substantial colocalization was observed between Dextran‐AF680 targeted cells (red) and CD206‐FITC+ cells (green). (C) Antibody‐dependent cellular phagocytosis (ADCP) occured in rapid, brief waves between Dextran‐AF680‐labeled macrophages and CD47 inhibitor‐treated tumor cells in a skinfold TNBC model. (D) CD47 inhibitor‐treated cancer cells (green) were engulfed by macrophages labeled with the pan‐macrophage marker F4/80‐PE (red), underscoring the highly phagocytic nature of macrophages (scale bars, 25 μm). (E) Intravital imaging of the robust phagocytic capacity of F4/80+ macrophages (red) toward CD47 inhibitor‐treated tumor cells (green).

Article Snippet: To evaluate the performance of the in vivo immunoscoring technique for the monitoring of anti‐CD47 immunotherapy efficacy, anti‐mouse CD47 monoclonal antibodies (specifically, anti‐CD47 mAbs, clone MIAP301, Rat IgG2a, κ, BioXcell, USA) and an IgG2a mAbs isotype control (clone 2A3, Rat IgG2a, κ, BioXcell, USA) were systemically and intravenously (IV) injected at a dosage of 10 mg/kg once a day for a duration of five consecutive days.

Techniques: Imaging, In Vivo, Injection, Labeling, Marker

Journal: Cancer Medicine

Article Title: In Vivo Immunoscoring Technique to Monitor Anti‐ CD47 Immunotherapy Efficacy in Triple‐Negative Breast Cancer

doi: 10.1002/cam4.72232

Figure Lengend Snippet: In vivo and in vitro validation of TNBC targeting by MCP1‐FITC. (A, B) Following intravenous injection of MCP1‐FITC, fluorescence signals in TNBC tissue peaked at 4 min and rapidly declined within 2 h ( n = 6, p = 0.0313). (C, D) Ex vivo imaging of dissected organs from subcutaneous xenograft models revealed a noticeable difference in fluorescence signal intensity between TNBC tumors and normal organs at 4 min after intravenous injection of MCP1‐FITC ( n = 6, p = 0.0095). (E, F) The average radiant efficiency of MCP1‐FITC in resected TNBC tissues was quantitively compared between the anti‐CD47 monoclonal antibody (mAb) and isotype control (ISO) treatment groups ( n = 16, p = 0.0499). (G) A significant difference in the M1/M2 ratio, based on the average radiant efficiency, was observed between anti‐CD47 mAb‐treated and ISO‐treated mice ( n = 12, p = 0.0022). (H) A pronounced shift in the population of MCP1‐FITC targeted cells was observed following repeated immunotherapy. (I, J) Significant differences in the M1/M2 ratio, based on the MCP‐1 FITC+ cells, is observed between anti‐CD47 mAb‐treated and ISO‐treated mice ( n = 16, p = 0.0104), ( n = 10, p = 0.0079). (K) A correlation analysis indicated that there is a strong correlation between the flow cytometry‐derived M1/M2 ratio and the imaging‐derived M1/M2 ratio. Data are presented as the mean ± standard deviation (SD).

Article Snippet: To evaluate the performance of the in vivo immunoscoring technique for the monitoring of anti‐CD47 immunotherapy efficacy, anti‐mouse CD47 monoclonal antibodies (specifically, anti‐CD47 mAbs, clone MIAP301, Rat IgG2a, κ, BioXcell, USA) and an IgG2a mAbs isotype control (clone 2A3, Rat IgG2a, κ, BioXcell, USA) were systemically and intravenously (IV) injected at a dosage of 10 mg/kg once a day for a duration of five consecutive days.

Techniques: In Vivo, In Vitro, Biomarker Discovery, Injection, Fluorescence, Tissue, Ex Vivo, Imaging, Xenograft Assay, Control, Analysis, Flow Cytometry, Derivative Assay, Standard Deviation