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Comprehensive Analysis of Drug Response using the FLICK Assay
Gavin A Birdsall1, Michael J Lee2
1Department of Systems Biology, UMass Chan Medical School.
Abstract:
For understanding drug efficacy, a critical need is to characterize the extent of drug-induced cell death. Efforts to quantify the level of drug-induced cell death are challenged by the existence of more than a dozen molecularly distinct forms of regulated death, each with its own activation timing and biochemical hallmark features. Furthermore, for some necrotic death subtypes, hallmark features are only observed transiently and are rapidly lost due to cell rupture. Thus, even when using a combination of death pathway-specific assays, it is challenging to accurately quantify the total amount of cell death or the relative contributions of each death subtype. Another issue is that many death-specific assays ignore how drugs affect cell proliferation, making it challenging to interpret if a drug-treated population is expanding or shrinking. The FLICK assay allows for quantification of the total level of cell death following stimulation in a manner that is specific to death but also largely agnostic to the type(s) of death activated. Additionally, the FLICK assay retains information about the total population size and cell proliferation rate. In this manuscript, we describe the basic use of the FLICK assay, how to troubleshoot this assay when using different types of biological material, and how to use the FLICK assay to quantify the contributions of each type of cell death to an observed drug response.
Insights
Quantifying drug-induced cell death is complex due to diverse mechanisms. The FLICK assay accurately measures total cell death and proliferation, overcoming limitations of traditional methods for drug efficacy studies.
Area of Science:
- Cell Biology
- Pharmacology
- Biochemistry
Background:
- Accurate quantification of drug-induced cell death is crucial for understanding drug efficacy.
- Existing methods face challenges due to the diversity of regulated cell death pathways and transient hallmark features.
- Many assays neglect the impact of drugs on cell proliferation, complicating data interpretation.
Purpose of the Study:
- To introduce and detail the FLICK assay for quantifying drug-induced cell death.
- To provide guidance on troubleshooting the FLICK assay across various biological materials.
- To demonstrate the utility of the FLICK assay in assessing contributions of distinct cell death types to drug responses.
Main Methods:
- The FLICK assay measures total cell death independent of specific death pathway activation.
- This method simultaneously quantifies cell death levels and population size/proliferation rates.
- The manuscript details the assay's application and troubleshooting for diverse biological samples.
Main Results:
- The FLICK assay offers a robust method for measuring total cell death.
- It provides insights into cell proliferation dynamics alongside cell death quantification.
- The assay enables the deconvolution of contributions from different cell death subtypes.
Conclusions:
- The FLICK assay is a versatile tool for comprehensive analysis of drug-induced cell death.
- It addresses key limitations of conventional assays, improving the assessment of drug efficacy.
- This method enhances the understanding of cell death mechanisms and their role in drug response.
