Characterisation of anticancer peptides at the single-cell level

Area of Science:

• Biomedical Engineering
• Pharmacology
• Cancer Research

Background:

• Chemotherapy efficacy is limited by diverse cellular responses.
• Anticancer peptides (ACPs) show promise but their effects on tumor heterogeneity are understudied.
• Addressing single-cell heterogeneity is crucial for developing effective cancer therapeutics.

Purpose of the Study:

• To develop and validate a microfluidic platform for analyzing single-cell drug responses.
• To investigate the effects of membranolytic ACPs on MCF-7 breast cancer cells at the single-cell level.
• To provide a tool for studying drug response heterogeneity in cancer.

Main Methods:

• A microfluidic device with hydrodynamic traps and microchambers was designed for single-cell isolation.
• Pneumatic valves were used to expose individual cells to different anticancer peptides (melittin, aurein 1.2, aurein 2.2).
• Calcein dye efflux was monitored to assess membrane integrity and cell viability.

Main Results:

• The microfluidic platform successfully captured and isolated hundreds of single MCF-7 cells.
• The study monitored the membrane integrity loss in single cells upon exposure to three different ACPs.
• Two imaging strategies were employed for in-depth mechanistic studies and high-throughput parallel analysis.

Conclusions:

• The developed microfluidic device enables the analysis of drug effects on large populations of single cells.
• This platform is adaptable for various cell types and drug screening applications.
• Future integration with other on-chip methods will elucidate the origins of heterogeneous drug responses.