Single-Cell Profiling Reveals Functional Heterogeneity and Serial Killing in Human Peripheral and Ex Vivo-Generated
Nikita Subedi1,2, Liesbeth Petronella Verhagen1,2, Paul de Jonge3
1Laboratory of Immunoengineering, Department of Biomedical Engineering, Eindhoven University of Technology, Groene Loper 5, Eindhoven, 5600 MB, The Netherlands.
Abstract:
Increasing evidence suggests that natural killer (NK) cells are composed of distinct functional subsets. This multifunctional role has made them an attractive choice for anticancer immunotherapy. A functional NK cell repertoire is generated through cellular education, resulting in a heterogeneous NK cell population with distinct capabilities responding to different stimuli. The application of a high-throughput droplet-based microfluidic platform allows monitoring of NK cell-target cell interactions at the single-cell level and in real-time. A variable response of single NK cells toward different target cells is observed, and a distinct population of NK cells (serial killers) capable of inducing multiple target lysis is identified. By assessing the cytotoxic dynamics, it is shown that single umbilical cord blood-derived CD34+ hematopoietic progenitor (HPC)-NK cells display superior antitumor cytotoxicity. With an integrated analysis of cytotoxicity and cytokine secretion, it is shown that target cell interactions augment cytotoxic as well as secretory behavior of NK cells. By providing an integrated assessment of NK cell functions by microfluidics, this study paves the way to further functionally characterize NK cells ultimately aimed to improve cancer immunotherapy.
Insights
Natural killer (NK) cells show varied responses to cancer cells. A microfluidic platform identified "serial killer" NK cells with superior tumor-killing ability, enhancing immunotherapy potential.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Natural killer (NK) cells are crucial for innate immunity and cancer surveillance.
- NK cell heterogeneity suggests distinct functional subsets, impacting their therapeutic potential.
- Understanding NK cell function at a single-cell level is key for optimizing immunotherapy.
Purpose of the Study:
- To investigate the functional heterogeneity of NK cells using a microfluidic platform.
- To identify specific NK cell subsets with enhanced cytotoxic capabilities against cancer.
- To analyze the integrated cytotoxic and secretory functions of NK cells in response to target cells.
Main Methods:
- Utilized a high-throughput droplet-based microfluidic platform for real-time, single-cell analysis of NK cell-target cell interactions.
- Assessed cytotoxic dynamics and quantified target cell lysis by individual NK cells.
- Integrated analysis of cytotoxicity and cytokine secretion from NK cells.
Main Results:
- Observed variable responses of single NK cells towards different target cells.
- Identified a distinct population of NK cells, termed 'serial killers', capable of multiple target lyses.
- Demonstrated superior antitumor cytotoxicity of umbilical cord blood-derived CD34+ hematopoietic progenitor (HPC)-NK cells.
- Showed that target cell interactions enhance both cytotoxic and secretory functions of NK cells.
Conclusions:
- Microfluidic analysis provides an integrated assessment of NK cell functions.
- Functional characterization of NK cell subsets can significantly improve anticancer immunotherapy strategies.
- HPC-NK cells represent a promising candidate for enhanced cancer immunotherapy due to their potent cytotoxic activity.
