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Microfluidic Co-Culture Models for Dissecting the Immune Response in in vitro Tumor Microenvironments
Published on: April 30, 2021
Machine learning-aided quantification of antibody-based cancer immunotherapy by natural killer cells in microfluidic
Saheli Sarkar1, Wenjing Kang, Songyao Jiang
1Department of Pharmaceutical Sciences, Northeastern University, 360 Huntington Avenue, Boston, MA, USA. t.konry@neu.edu.
Abstract:
Natural killer (NK) cells have emerged as an effective alternative option to T cell-based immunotherapies, particularly against liquid (hematologic) tumors. However, the effectiveness of NK cell therapy has been less than optimal for solid tumors, partly due to the heterogeneity in target interaction leading to variable anti-tumor cytotoxicity. This paper describes a microfluidic droplet-based cytotoxicity assay for quantitative comparison of immunotherapeutic NK-92 cell interaction with various types of target cells. Machine learning algorithms were developed to assess the dynamics of individual effector-target cell pair conjugation and target death in droplets in a semi-automated manner. Our results showed that while short contacts were sufficient to induce potent killing of hematological cancer cells, long-lasting stable conjugation with NK-92 cells was unable to kill HER2+ solid tumor cells (SKOV3, SKBR3) significantly. NK-92 cells that were engineered to express FcγRIII (CD16) mediated antibody-dependent cellular cytotoxicity (ADCC) selectively against HER2+ cells upon addition of Herceptin (trastuzumab). The requirement of CD16, Herceptin and specific pre-incubation temperature served as three inputs to generate a molecular logic function with HER2+ cell death as the output. Mass proteomic analysis of the two effector cell lines suggested differential changes in adhesion, exocytosis, metabolism, transport and activation of upstream regulators and cytotoxicity mediators, which can be utilized to regulate specific functionalities of NK-92 cells in future. These results suggest that this semi-automated single cell assay can reveal the variability and functional potency of NK cells and may be used to optimize immunotherapeutic efficacy for preclinical analyses.
Insights
This study introduces a microfluidic assay to evaluate natural killer (NK) cell therapy for solid tumors. The assay reveals NK-92 cell limitations against HER2+ tumors, but shows potential for engineered cells via antibody-dependent cellular cytotoxicity.
Area of Science:
- Immunology
- Biotechnology
- Cancer Research
Background:
- Natural killer (NK) cells show promise in immunotherapy, especially for hematologic cancers.
- NK cell therapy efficacy is limited in solid tumors due to variable anti-tumor cytotoxicity.
- Target interaction heterogeneity affects NK cell effectiveness against diverse cancer types.
Purpose of the Study:
- To develop and validate a microfluidic droplet-based assay for quantitative assessment of NK-92 cell interactions with target cells.
- To investigate the dynamics of effector-target cell conjugation and cytotoxicity in individual cell pairs.
- To compare NK-92 cell efficacy against hematologic and solid tumor cells, and explore strategies for enhancing solid tumor targeting.
Main Methods:
- A microfluidic droplet-based assay was employed for single-cell analysis of NK-92 cell interactions.
- Machine learning algorithms were utilized for semi-automated assessment of cell conjugation and target cell death.
- NK-92 cells were engineered to express FcγRIII (CD16) to investigate antibody-dependent cellular cytotoxicity (ADCC).
- Mass proteomic analysis was performed on effector cell lines.
Main Results:
- Short NK-92 cell contact sufficed for potent killing of hematologic cancer cells.
- Prolonged NK-92 cell conjugation did not significantly kill HER2+ solid tumor cells (SKOV3, SKBR3).
- Engineered NK-92 cells expressing CD16 mediated selective ADCC against HER2+ cells with Herceptin.
- A molecular logic function was established linking CD16, Herceptin, and temperature to HER2+ cell death.
- Proteomic analysis revealed differential changes in effector cell lines related to adhesion, metabolism, and cytotoxicity mediators.
Conclusions:
- The developed microfluidic assay enables quantitative, single-cell analysis of NK cell potency and variability.
- NK-92 cells demonstrate limitations against HER2+ solid tumors, highlighting the need for enhanced therapeutic strategies.
- Engineered NK-92 cells utilizing ADCC show potential for targeted HER2+ solid tumor therapy.
- Understanding effector cell proteomic profiles can guide optimization of NK cell-based immunotherapies for improved efficacy in preclinical settings.
