Microfluidic single-cell drug screening: toward personalized precision therapy in chronic myeloid leukemia
Hwisu Jeon1, Yukyung Park2, Soo-Hyun Kim3,4
1Department of Mechanical Engineering, Ulsan National Institute of Science and Technology (UNIST), 50 UNIST-Gil, Ulsan 44919, Republic of Korea. tskim@unist.ac.kr.
Abstract:
Chronic myeloid leukemia (CML) treatment employs several FDA-approved BCR::ABL1 tyrosine kinase inhibitors (TKIs) with distinct efficacy and side effects influenced by patient-specific factors. This study introduces a microfluidic cell culture array for the comparative analysis of six BCR::ABL1 TKIs, namely imatinib, nilotinib, bosutinib, ponatinib, dasatinib, and asciminib, using CML-related cell lines. The device provides a continuous, chemostat-like microfluidic environment that enables quantitative drug sensitivity scoring. The microchambers for cell culture notably offer advantages for single-cell imaging of suspension leukemia cells, which tend to aggregate in conventional culture platforms. This system supports the detailed characterization of cell viability across various TKI types and concentrations, yielding comprehensive mathematical metrics to assess relative drug efficacy. In this study, we compared drug responses in K562 and Ba/F3 BCR::ABL1 cell lines, including the T315I mutant variant, and specifically demonstrated that Ba/F3 cells harboring the T315I mutation exhibit resistance to the first- and second-generation TKIs, responding only to ponatinib and asciminib. We further validated the device with a CML patient-derived bone marrow sample, requiring only minimal adjustments to the experimental conditions. The proposed microfluidic single-cell-based screening array could refine treatment regimens and advance personalized medicine in CML.
Insights
A new microfluidic device enables precise testing of multiple BCR::ABL1 tyrosine kinase inhibitors (TKIs) for chronic myeloid leukemia (CML). This technology aids in selecting the most effective TKI for individual patients, advancing personalized CML treatment.
Area of Science:
- Oncology
- Biotechnology
- Microfluidics
Background:
- Chronic myeloid leukemia (CML) treatment relies on BCR::ABL1 tyrosine kinase inhibitors (TKIs), but efficacy varies due to patient-specific factors.
- Conventional cell culture methods struggle with leukemia cell aggregation, hindering detailed analysis.
- Personalized medicine approaches are needed to optimize TKI selection for CML patients.
Purpose of the Study:
- To develop and validate a microfluidic cell culture array for comparative analysis of six BCR::ABL1 TKIs.
- To enable quantitative drug sensitivity scoring and single-cell imaging for CML cell lines and patient samples.
- To assess the efficacy of different TKIs, including against resistant mutations like T315I.
Main Methods:
- A microfluidic device creating a chemostat-like environment for continuous cell culture.
- Comparative drug sensitivity analysis of imatinib, nilotinib, bosutinib, ponatinib, dasatinib, and asciminib.
- Single-cell imaging and viability assessment in K562, Ba/F3 BCR::ABL1 (including T315I mutant), and patient-derived CML cells.
Main Results:
- The microfluidic system successfully quantified drug sensitivity and enabled single-cell analysis of leukemia cells.
- Ba/F3 cells with the T315I mutation showed resistance to first- and second-generation TKIs, responding only to ponatinib and asciminib.
- The device was validated using a CML patient-derived bone marrow sample with minimal experimental adjustments.
Conclusions:
- The microfluidic single-cell screening array offers a robust platform for evaluating BCR::ABL1 TKIs in CML.
- This technology can refine treatment strategies by enabling precise TKI efficacy assessment.
- The system holds significant potential for advancing personalized medicine in CML treatment.
