Journal: Scientific Reports

Article Title: EGFR and HER3 expression in circulating tumor cells and tumor tissue from non-small cell lung cancer patients

doi: 10.1038/s41598-019-43678-6

Figure Lengend Snippet: Protein expression and frequency distribution in primary and metastatic lung cancer tissue. EGFR protein immunohistochemistry ( A ) and HER3 protein immunohistochemistry ( B ) was performed on tissue microarray from primary NSCLC tumors and brain metastases. Representative strong (left), intermediate (center) and negative (right) staining are shown in 200x magnification (Zeiss Axiovision).

Article Snippet: For HER3 we tested two different antibodies; the biotinylated human anti-HER3 antibody (clone REA508; Miltenyi Biotec, Bergisch Gladbach, Germany) and the biotinylated human anti-HER-3 antibody (clone 1B4C3; BioLegend, San Diego, California).

Techniques: Expressing, Immunohistochemistry, Microarray, Staining

Journal: Scientific Reports

Article Title: EGFR and HER3 expression in circulating tumor cells and tumor tissue from non-small cell lung cancer patients

doi: 10.1038/s41598-019-43678-6

Figure Lengend Snippet: EGFR ( A ) and HER3 ( B ) expression distribution in primary lung, lymph node and brain metastatic tissue and association between HER3 protein expression in primary and clinicopathological parameters ( C ). HER3 is more frequently expressed in brain metastasis of oligo-brain metastatic patients (p = 0.028) compared to patients with other metastatic sites. HER3 expression in primary NSCLC tumors is significantly associated with a decreased time to metastatic progression (p = 0.006; log-rank test) ( D ) and decreased relapse-free survival time (p = 0.013; log-rank test) ( E ).

Article Snippet: For HER3 we tested two different antibodies; the biotinylated human anti-HER3 antibody (clone REA508; Miltenyi Biotec, Bergisch Gladbach, Germany) and the biotinylated human anti-HER-3 antibody (clone 1B4C3; BioLegend, San Diego, California).

Techniques: Expressing

Journal: Scientific Reports

Article Title: EGFR and HER3 expression in circulating tumor cells and tumor tissue from non-small cell lung cancer patients

doi: 10.1038/s41598-019-43678-6

Figure Lengend Snippet: CTC positivity rates in NSCLC patients (n = 45). ( A ) Significantly higher detection rates when combining the methods in comparison to either single use CellSearch (p = 0.0023) or magnetic cell separation with EGFR/HER3 (p = 0.0109). ( B ) CTC enumeration detected by CellSearch and MACS.

Article Snippet: For HER3 we tested two different antibodies; the biotinylated human anti-HER3 antibody (clone REA508; Miltenyi Biotec, Bergisch Gladbach, Germany) and the biotinylated human anti-HER-3 antibody (clone 1B4C3; BioLegend, San Diego, California).

Techniques: Comparison, Magnetic Cell Separation

Journal: Scientific Reports

Article Title: EGFR and HER3 expression in circulating tumor cells and tumor tissue from non-small cell lung cancer patients

doi: 10.1038/s41598-019-43678-6

Figure Lengend Snippet: Representative images of two Circulating Tumor Cell cluster (DAPI + /CD45 − /CK + ) isolated via magnetic cell separation with HER3-protein antibody in one NSCLC patient. Heterogeneous MET expression was found within the cluster as well as CD45-positive leukocytes.

Article Snippet: For HER3 we tested two different antibodies; the biotinylated human anti-HER3 antibody (clone REA508; Miltenyi Biotec, Bergisch Gladbach, Germany) and the biotinylated human anti-HER-3 antibody (clone 1B4C3; BioLegend, San Diego, California).

Techniques: Isolation, Magnetic Cell Separation, Expressing

Journal: American Journal of Cancer Research

Article Title: CAN017, a novel anti-HER3 antibody, exerted great potency in mouse avatars of esophageal squamous cell carcinoma with NRG1 as a biomarker

doi:

Figure Lengend Snippet: Tolerance and efficacy of CAN017 in ES0199 and ES0195 PDX models. A. Body weights of mice during treatment. Data were presented as means ± SDs. B. Tumor growth curves showed the in vivo activity of CAN017 in ES0199 and ES0195 PDX models treated with vehicle control, hIgG control (20 mg/kg) and CAN017 (20 mg/kg) every 3 days. Data were indicated as means ± SDs. The anti-tumor activity was depicted by TGI with 105.92% and 44.86% in ES0199 and ES0195 PDX models, respectively. C. Kaplan-Meier graph showed the probability of tumor progression-free mice in ES0199 and ES0195 PDX models during the treatment. **P < 0.01; ns, not significant by log-rank test.

Article Snippet: Reagents CAN017 antibody (Anti-HER3-hIgG1/κ) was kindly provided by CANBridge Life Sciences Ltd. (Beijing, China) and dissolved in saline at a stock concentration of 50 mg/mL, and then stored at 2-8°C until used.

Techniques: In Vivo, Activity Assay, Control

Journal: American Journal of Cancer Research

Article Title: CAN017, a novel anti-HER3 antibody, exerted great potency in mouse avatars of esophageal squamous cell carcinoma with NRG1 as a biomarker

doi:

Figure Lengend Snippet: Efficacy of CAN017 in large ESCC PDX models in cohort 1. A. Tumor growth curves showed the in vivo activity of 22 ESCC PDX models treated with vehicle control, hIgG control (20 mg/kg) and CAN017 (20 mg/kg) every 3 days. Data were expressed as means ± SDs for at less five mice in each group. The anti-tumor activity was depicted by TGI described in each picture. B. Waterfall plot displayed the different response to CAN017 of 24 PDX models in cohort 1. Dotted line indicated the TGI of 70%. C. Comparison of tumor growth inhibition between hIgG group and CAN017 group. ***P < 0.001 analyzed by unpaired two-tailed t test.

Article Snippet: Reagents CAN017 antibody (Anti-HER3-hIgG1/κ) was kindly provided by CANBridge Life Sciences Ltd. (Beijing, China) and dissolved in saline at a stock concentration of 50 mg/mL, and then stored at 2-8°C until used.

Techniques: In Vivo, Activity Assay, Control, Comparison, Inhibition, Two Tailed Test

Journal: American Journal of Cancer Research

Article Title: CAN017, a novel anti-HER3 antibody, exerted great potency in mouse avatars of esophageal squamous cell carcinoma with NRG1 as a biomarker

doi:

Figure Lengend Snippet: Correlation between HER3 and NRG1 expression and the efficacy of CAN017 in 24 PDX models in cohort 1. A. Pearson correlation plots of the HER3 and NRG1 protein expressions evaluated by IHC in FFPE tumors and the efficacy of CAN017 in PDX models (n = 24). The linear regression was shown by a solid line, and 95% confidence interval (CI) of the values fitted by linear regression was shown by dark dotted lines. R2, the coefficient of determination. B. Pearson correlation plots of the HER3 and NRG1 mRNA expression quantified by RNAseq in tumors and the efficacy of CAN017 in PDX models (n = 24). The linear regression was shown by a solid line, and 95% CI of the values fitted by linear regression was shown by dark dotted lines. C. Pearson correlation plots of the NRG1 mRNA expression quantified by RT-PCR in FFPE tumors and the efficacy of CAN017 in PDX models (n = 24). The linear regression was shown by a solid line, and 95% CI of the values fitted by linear regression was shown by dark dotted lines. D. ROC curve analysis for the NRG1 ΔCt threshold to predict the response of CAN017. AUC, area under curve. E. Comparison of the efficacy of CAN017 between NRG1-high and NRG1-low PDX models. ****P < 0.0001 analyzed by unpaired two-tailed t test.

Article Snippet: Reagents CAN017 antibody (Anti-HER3-hIgG1/κ) was kindly provided by CANBridge Life Sciences Ltd. (Beijing, China) and dissolved in saline at a stock concentration of 50 mg/mL, and then stored at 2-8°C until used.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Comparison, Two Tailed Test

Journal: American Journal of Cancer Research

Article Title: CAN017, a novel anti-HER3 antibody, exerted great potency in mouse avatars of esophageal squamous cell carcinoma with NRG1 as a biomarker

doi:

Figure Lengend Snippet: Efficacy of CAN017 prospectively guided by NRG1 expression in cohort 2. A. Tumor growth curves showed the in vivo activity of 5 ESCC PDX models treated with vehicle control and CAN017 (20 mg/kg). Data were expressed as means ± SDs for at less five mice in each group. The anti-tumor activity was depicted by TGI as described in each picture. B. NRG1 mRNA expression levels of 5 ESCC tumors and the efficacy of CAN017 on PDX models. Blue dotted lines showed the threshold of NRG1 ΔCt and the TGI of 70%, respectively.

Article Snippet: Reagents CAN017 antibody (Anti-HER3-hIgG1/κ) was kindly provided by CANBridge Life Sciences Ltd. (Beijing, China) and dissolved in saline at a stock concentration of 50 mg/mL, and then stored at 2-8°C until used.

Techniques: Expressing, In Vivo, Activity Assay, Control

Journal: BMC Cancer

Article Title: The prognostic significance of human epidermal growth factor receptor family protein expression in operable pancreatic cancer

doi: 10.1186/s12885-016-2889-6

Figure Lengend Snippet: pHER3 expression in AsPC-1 cells and the inhibitory effect of anti-HER3 monoclonal antibody (3D4). a . pHER3 was overexpressed in the AsPC-1 cells. Both 3D4 and trastuzumab significantly down-regulated the expression of pHER3, and two drug combination showed synergistic inhibitory effect. b . 3D4 significantly inhibited cellular proliferation at concentrations equal to 20 ug/ml or 50 μg/ml ( P < 0.05)

Article Snippet: Cells were then treated with serially diluted anti-HER3 monoclonal antibody (clone 3D4, Beijing Cotimes Biotech Co., Ltd.) for 48 h, after which 10 μl of a 5 mg/ml stock of MTT (Sigma) was added to the wells.

Techniques: Expressing

Journal: BMC Cancer

Article Title: The prognostic significance of human epidermal growth factor receptor family protein expression in operable pancreatic cancer

doi: 10.1186/s12885-016-2889-6

Figure Lengend Snippet: The expression of HER1 - HER4 in different subgroups

Article Snippet: Cells were then treated with serially diluted anti-HER3 monoclonal antibody (clone 3D4, Beijing Cotimes Biotech Co., Ltd.) for 48 h, after which 10 μl of a 5 mg/ml stock of MTT (Sigma) was added to the wells.

Techniques: Expressing

Journal: BMC Cancer

Article Title: The prognostic significance of human epidermal growth factor receptor family protein expression in operable pancreatic cancer

doi: 10.1186/s12885-016-2889-6

Figure Lengend Snippet: The cancer-specific survival and HER1 - HER4 expression

Article Snippet: Cells were then treated with serially diluted anti-HER3 monoclonal antibody (clone 3D4, Beijing Cotimes Biotech Co., Ltd.) for 48 h, after which 10 μl of a 5 mg/ml stock of MTT (Sigma) was added to the wells.

Techniques:

Journal: BMC Cancer

Article Title: The prognostic significance of human epidermal growth factor receptor family protein expression in operable pancreatic cancer

doi: 10.1186/s12885-016-2889-6

Figure Lengend Snippet: Cox’s multiple regression analysis for survival time

Article Snippet: Cells were then treated with serially diluted anti-HER3 monoclonal antibody (clone 3D4, Beijing Cotimes Biotech Co., Ltd.) for 48 h, after which 10 μl of a 5 mg/ml stock of MTT (Sigma) was added to the wells.

Techniques: Histopathology

Journal: Cells

Article Title: Inhibition of HER Receptors Reveals Distinct Mechanisms of Compensatory Upregulation of Other HER Family Members: Basis for Acquired Resistance and for Combination Therapy

doi: 10.3390/cells10020272

Figure Lengend Snippet: HER single knockdown leads to the counter-upregulation of the respective other HER family member, and combined HER2/HER3 knockdown leads to enhanced antitumor effects. ( A ) SKOV3 cells were transfected with siRNAs as indicated in the figures and analyzed for mRNA levels of HER2 (left) and HER3 (right). ( B ) Alterations of HER2 and HER3 protein levels, as determined by flow cytometry. ( C ) Anti-proliferative and ( D ) pro-apoptotic effects upon siRNA-mediated single or double knockdown of HER2 and/or HER3.

Article Snippet: The HER3 antibody was obtained from GeneTex (Irvine, CA, USA) and diluted 1:500 in the same buffer.

Techniques: Knockdown, Transfection, Flow Cytometry

Journal: Cells

Article Title: Inhibition of HER Receptors Reveals Distinct Mechanisms of Compensatory Upregulation of Other HER Family Members: Basis for Acquired Resistance and for Combination Therapy

doi: 10.3390/cells10020272

Figure Lengend Snippet: Systemic treatment of mice bearing s.c. SKOV3 tumor xenografts with PEI/siRNA complexes for HER knockdown. ( A ) Anti-tumor effects of HER single or double knockdown by systemic (i.p.) injection of PEI/siRNA complexes. ( B ) Immunohistochemical analysis of HER2 protein levels in tumors from different treatment groups. Representative examples of stained slides without counterstain are shown. ( C ) Analysis of tumor lysates for HER3 protein levels.

Article Snippet: The HER3 antibody was obtained from GeneTex (Irvine, CA, USA) and diluted 1:500 in the same buffer.

Techniques: Knockdown, Injection, Immunohistochemical staining, Staining

Journal: Cells

Article Title: Inhibition of HER Receptors Reveals Distinct Mechanisms of Compensatory Upregulation of Other HER Family Members: Basis for Acquired Resistance and for Combination Therapy

doi: 10.3390/cells10020272

Figure Lengend Snippet: Role of miR143 in the HER2/HER3 regulation. ( A ) Effect of miR-143 transfection on HER3 protein levels. ( B ) HER3 is a direct target of miR-143 as determined in a luciferase reporter assay. ( C ) Upon siRNA-mediated HER2 knockdown, a marked reduction of miR-143 levels is observed. ( D ) MiR-143 levels in the tumor xenografts from the PEI/siRNA therapy experiment.

Article Snippet: The HER3 antibody was obtained from GeneTex (Irvine, CA, USA) and diluted 1:500 in the same buffer.

Techniques: Transfection, Luciferase, Reporter Assay, Knockdown

Journal: Cells

Article Title: Inhibition of HER Receptors Reveals Distinct Mechanisms of Compensatory Upregulation of Other HER Family Members: Basis for Acquired Resistance and for Combination Therapy

doi: 10.3390/cells10020272

Figure Lengend Snippet: Effect of SATB1 in the HER3/HER2 regulation network. ( A ) Alterations in SATB1 mRNA levels upon knockdown of HER2 or HER3. ( B ) SiRNA-mediated SATB1 knockdown leads to increased HER2 mRNA levels. ( C ) SATB1 overexpression decreases HER2 mRNA levels. ( D ) SATB1 levels in the tumor xenografts from the PEI/siRNA therapy experiment, as determined by immunohistochemistry.

Article Snippet: The HER3 antibody was obtained from GeneTex (Irvine, CA, USA) and diluted 1:500 in the same buffer.

Techniques: Knockdown, Over Expression, Immunohistochemistry

Journal: Cells

Article Title: Inhibition of HER Receptors Reveals Distinct Mechanisms of Compensatory Upregulation of Other HER Family Members: Basis for Acquired Resistance and for Combination Therapy

doi: 10.3390/cells10020272

Figure Lengend Snippet: Effects of TX-1-85-1 or trastuzumab (Herceptin) on the HER regulation network. SKOV3 cells were analyzed for ( A ) HER2 levels upon treatment with the inhibitor of HER3 phosphorylation, TX-1-85-1, or treated with Herceptin and analyzed for ( B ) HER3 and ( C ) miR-143 levels.

Article Snippet: The HER3 antibody was obtained from GeneTex (Irvine, CA, USA) and diluted 1:500 in the same buffer.

Techniques: Phospho-proteomics

Journal: Cells

Article Title: Inhibition of HER Receptors Reveals Distinct Mechanisms of Compensatory Upregulation of Other HER Family Members: Basis for Acquired Resistance and for Combination Therapy

doi: 10.3390/cells10020272

Figure Lengend Snippet: Schematic overview of the network of HER counter-(up)regulation processes (details: see text). ( A ) Effects upon the PEI/siHER2-mediated knockdown or the trastuzumab (Herceptin)-mediated inhibition of HER2, leading to HER3 upregulation. ( B ) HER3 knockdown or TX1-85-1-mediated inhibition with subsequent HER2 upregulation. Red arrows indicate the effects upon the respective inhibition or knockdown.

Article Snippet: The HER3 antibody was obtained from GeneTex (Irvine, CA, USA) and diluted 1:500 in the same buffer.

Techniques: Knockdown, Inhibition