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Visualization of natural killer cell-mediated killing of cancer cells at single-cell resolution in live zebrafish
Hongmei Yang1, Hao Jia1, Qi Zhao2
1Faculty of Health Sciences, University of Macau, Taipa, Macao SAR, China.
Abstract:
Tumor immunotherapy has been an important advancement in cancer treatment in recent years. Compared with T cell-based therapy, natural killer (NK) cell-based therapy does not require human leukocyte antigen matching and has fewer side effects; thus, NK cell therapy has gradually attracted the attention of researchers and clinicians. Reliable and effective animal models are essential for evaluating the effects of NK cell therapy. NK cells kill cancer cells mainly through apoptosis. In this study, we first established a 3D coculture model using fluorescence resonance energy transfer (FRET)-based lung or breast cancer cells and tdTomato-labeled NK cells. We observed that cancer cells changed from green to blue when undergoing apoptosis induced by red NK cells. We then coinjected these green cancer cells with red NK cells into zebrafish to visualize the interaction between them and the killing process of NK cells against cancer cells in real-time and at single-cell resolution in circulation. Using this model, we found that NK cells can quickly kill cancer cells in zebrafish circulation in 40 min and the caspase-3 can be activated in 5-10 min. This FRET-based zebrafish tumor model can serve as a powerful in vivo tool that can facilitate the development of NK cell-based therapy. More importantly, cancer cells from cancer patients can be labeled with our apoptotic biosensor and then transplanted into zebrafish to evaluate the sensitivity of the cancer cells to NK cells to help clinicians make treatment plans that can benefit patients.
Insights
Natural killer (NK) cell therapy shows promise for cancer treatment. Researchers developed a novel zebrafish model to visualize NK cells killing cancer cells in real-time, aiding therapy development.
Area of Science:
- Oncology
- Immunology
- Biotechnology
Background:
- Tumor immunotherapy, particularly natural killer (NK) cell therapy, is a rapidly advancing field in cancer treatment.
- NK cell therapy offers advantages over T cell therapy, including no requirement for human leukocyte antigen matching and fewer side effects.
- Effective animal models are crucial for assessing NK cell therapy efficacy.
Purpose of the Study:
- To establish and validate a novel in vivo model for real-time visualization and analysis of NK cell-mediated cancer cell apoptosis.
- To evaluate the dynamics of NK cell interaction with cancer cells and the kinetics of apoptosis induction.
- To create a tool for assessing patient-derived cancer cell sensitivity to NK cell therapy for personalized treatment planning.
Main Methods:
- Development of a 3D co-culture system using fluorescence resonance energy transfer (FRET)-based cancer cells and tdTomato-labeled NK cells.
- Establishment of a zebrafish model by co-injecting FRET-based cancer cells and red NK cells for in vivo imaging.
- Real-time, single-cell resolution tracking of NK cell-cancer cell interactions and apoptosis in zebrafish circulation.
Main Results:
- Observed cancer cell apoptosis via a color change (green to blue) in the FRET-based co-culture system.
- Demonstrated rapid killing of cancer cells by NK cells in zebrafish circulation within 40 minutes.
- Showed caspase-3 activation occurring within 5-10 minutes of NK cell interaction.
Conclusions:
- The FRET-based zebrafish model provides a powerful in vivo platform for studying NK cell therapy.
- This model facilitates the real-time evaluation of NK cell activity and cancer cell apoptosis.
- The model can be adapted to assess patient-derived cancer cell sensitivity to NK cells, aiding clinical treatment decisions.
