Quinacrine-mediated detection of intracellular ATP

Area of Science:

• Immunology
• Oncology
• Cell Biology

Background:

• Antineoplastic agents can induce immunogenic cell death (ICD), a process crucial for initiating anti-cancer immune responses.
• ICD involves the release of danger-associated molecular patterns, such as adenosine triphosphate (ATP), into the tumor microenvironment.
• Extracellular ATP signals through purinergic P2R receptors, promoting the recruitment and activation of immune cells to fight cancer.

Purpose of the Study:

• To develop and validate a fluorescence microscopy-based protocol for quantifying ATP secretion associated with ICD.
• To establish a high-throughput screening method for identifying novel ICD-inducing antineoplastic agents.
• To offer a cost-efficient alternative to traditional ATP detection methods.

Main Methods:

• Development of a fluorescence microscopy assay to measure extracellular ATP release from cells undergoing ICD.
• Adaptation of the assay for high-throughput screening applications.
• Comparison of the fluorescence microscopy method with conventional luciferase-based ATP detection.

Main Results:

• The developed protocol enables accurate quantification of ICD-associated ATP secretion.
• The fluorescence microscopy method is amenable to high-throughput screening, facilitating the evaluation of multiple compounds.
• This approach is more cost-efficient than traditional luciferase-based assays and allows for simultaneous assessment of cellular morphology.

Conclusions:

• A novel, cost-effective fluorescence microscopy protocol for quantifying ICD-associated ATP secretion has been established.
• This method supports high-throughput screening for immunogenic cell death inducers, aiding in the discovery of new cancer therapies.
• The protocol's ability to assess cellular morphology alongside ATP release provides a more comprehensive understanding of ICD induction.