Related Experiment Video
Updated: Jan 14, 2026
Assessment of Human Natural Killer Cell Events Driven by FcγRIIIa Engagement in the Presence of Therapeutic Antibodies
Published on: May 22, 2020
Overcoming NK cell resistance in triple-negative breast cancer via adcc with a humanized anti-CD147 antibody
Thanathat Pamonsupornwichit1, Kanyarat Thongheang1,2, Nuchjira Takheaw2,3
1Center of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand.
Abstract:
Triple-negative breast cancer (TNBC) is an aggressive and clinically challenging subtype defined by the absence of estrogen receptor, progesterone receptor, and HER2 amplification, resulting in poor prognosis and limited therapeutic options. Targeting alternative molecular pathways is urgently needed to overcome resistance and improve patient outcomes. CD147 has emerged as surface marker associated with tumor progression and immune evasion. In this study, CD147 and MHC class I-a key inhibitory ligand for natural killer cells-were analyzed in breast cancer cell lines (MCF7, MDA-MB-453, MDA-MB-231, and HCC38) using flow cytometry. The therapeutic efficacy of a humanized anti-CD147 monoclonal antibody (HuM6-1B9) was evaluated for its capacity to potentiate antibody-dependent cellular cytotoxicity (ADCC). HuM6-1B9 demonstrated the strong binding to MDA-MB-231 (KD = 4.982 nM) and HCC38 (KD = 4.523 nM), which are representative TNBC cell lines. In 3D spheroid models, HuM6-1B9 significantly enhanced PBMC-mediated ADCC, leading to a marked reduction in TNBC spheroid viability. Co-culture of CFSE-labeled MDA-MB-231 and HCC38 cells with primary NK cells confirmed robust ADCC, achieving 50% and 70% cytotoxicity, respectively, despite high MHC class I expression. Live-cell imaging demonstrated caspase-3/7 activation consistent with apoptosis in NK-targeted tumor cells, while CD107a degranulation and IFN-γ secretion confirmed the functional contribution of HuM6-1B9 to ADCC enhancement. Importantly, HuM6-1B9 did not promote migration or invasion in MDA-MB-231 cells, supporting its safety profile regarding metastasis. Collectively, these findings establish HuM6-1B9 as a promising immunotherapeutic candidate that overcomes immune resistance and selectively eliminates TNBC cells through ADCC without enhancing metastatic potential. By integrating mechanistic assays of NK cytotoxicity, apoptosis, and 3D tumor spheroids, this study provides clinically relevant insights underscoring the translational potential of HuM6-1B9 in TNBC immunotherapy.
Insights
A new antibody, HuM6-1B9, effectively targets triple-negative breast cancer (TNBC) by enhancing natural killer cell activity. This immunotherapy shows promise in eliminating TNBC cells without increasing metastasis, offering a potential new treatment option.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Triple-negative breast cancer (TNBC) presents a significant clinical challenge due to its aggressive nature and limited therapeutic options.
- CD147 is identified as a surface marker linked to tumor progression and immune evasion in TNBC.
- Overcoming resistance and improving patient outcomes necessitate the exploration of novel molecular targets and therapeutic strategies.
Purpose of the Study:
- To investigate the expression of CD147 and MHC class I in TNBC cell lines.
- To evaluate the therapeutic efficacy of a humanized anti-CD147 monoclonal antibody (HuM6-1B9) in potentiating antibody-dependent cellular cytotoxicity (ADCC).
- To assess the safety profile of HuM6-1B9 concerning tumor cell migration and invasion.
Main Methods:
- Flow cytometry was used to analyze CD147 and MHC class I expression in breast cancer cell lines.
- The binding affinity of HuM6-1B9 to TNBC cell lines (MDA-MB-231 and HCC38) was determined.
- 3D spheroid models and co-cultures with primary NK cells were employed to assess HuM6-1B9-mediated ADCC, cytotoxicity, apoptosis, and cytokine secretion.
- Cell migration and invasion assays were performed to evaluate the antibody's effect on metastatic potential.
Main Results:
- HuM6-1B9 exhibited strong binding affinity to representative TNBC cell lines, MDA-MB-231 and HCC38.
- The antibody significantly enhanced PBMC-mediated ADCC in 3D spheroid models, reducing TNBC spheroid viability.
- Robust ADCC was observed in co-cultures with NK cells, leading to significant tumor cell killing despite high MHC class I expression.
- HuM6-1B9 induced apoptosis in targeted tumor cells and promoted NK cell degranulation and IFN-γ secretion, confirming ADCC enhancement.
- Importantly, HuM6-1B9 did not increase the migration or invasion of MDA-MB-231 cells.
Conclusions:
- HuM6-1B9 is a promising immunotherapeutic candidate for TNBC, effectively overcoming immune resistance.
- The antibody selectively eliminates TNBC cells through enhanced ADCC mediated by NK cells.
- HuM6-1B9 demonstrates a favorable safety profile by not promoting metastasis, highlighting its translational potential in TNBC immunotherapy.
More Related Videos
13:19Quantifying Antibody-Dependent Cellular Cytotoxicity in a Tumor Spheroid Model: Application for Drug Discovery
Published on: April 26, 2024
13:59High-Content Screening Assay for the Identification of Antibody-Dependent Cellular Cytotoxicity Modifying Compounds
Published on: August 18, 2023