Spatial-Temporal Multiomics Profiling of Living Single Cells on Chip for Cancer Therapy

Single-cell molecular profiling has emerged as a cutting-edge technology in biological research, playing a pivotal role in elucidating fundamental life processes and cellular heterogeneity. However, traditional methods generally provide end point measurements and require cell disruption, posing challenges for long-term monitoring of living cell events, e.g., cancer therapy. Tumor Treating Fields (TTFields), a noninvasive therapeutic modality, exert inhibitory effects on tumor growth and invasion by applying low-intensity intermediate-frequency alternating electric fields to tumor regions, whose molecular mechanism on cancer physiology remains poorly understood. In this study, we propose a multifunctional microchip capable of continuous in situ monitoring of molecular signatures in living single cells. The platform integrates spatial cell positioning, single-cellular multimolecule tracking with surface-enhanced Raman scattering (SERS), and TTFields treatment functionalities on a microchip. Using multiplexed analysis of liver cancer cells treated with TTFields and chemotherapy drugs, we implement spatial-temporal multiomics profiling for cancer therapy. Temporal tracking of critical molecular events associated with glycolysis, genetic stability, and membrane integrity reveals significant differences between the groups with and without TTFields treatment. Causal network analysis of molecular profiles uncovers the underlying mechanisms through which TTFields modulate oncogenic pathways. Our work demonstrates the effectiveness of the proposed microchip platform for real-time monitoring of living cell molecular dynamics, offering a powerful tool for precision cancer therapy.