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Detecting the Senescence-Associated Secretory Phenotype (SASP) by High Content Microscopy Analysis
Priya Hari1, Juan Carlos Acosta2
1Edinburgh Cancer Research UK Centre, Institute of Genetics and Molecular Medicine, University of Edinburgh, Crewe Road South, Edinburgh, EH4 2XR, UK.
Methods in Molecular Biology (Clifton, N.J.)
|November 5, 2016
Summary
Senescent cells secrete proteins influencing aging and cancer. High-content analysis effectively characterizes these senescence-associated secretory phenotypes (SASP) and identifies regulators for potential therapeutic manipulation.
Area of Science:
- Cellular senescence
- Cancer biology
- Molecular biology
Background:
- Senescent cells secrete proteins (SASP) impacting aging and tumor microenvironments.
- Understanding SASP regulators is crucial for manipulating cellular senescence.
- High-throughput methods enable efficient characterization and screening of SASP.
Purpose of the Study:
- To characterize cellular phenotypes modulated by Oncogene-Induced Senescence (OIS) regulators.
- To utilize high-content analysis for measuring senescence markers, including SASP.
- To establish a screening method for identifying regulators of SASP during OIS.
Main Methods:
- Employing Widefield High-Content Analysis Systems for phenotypic characterization.
- Measuring multiple senescence markers, including SASP components.
- Applying the system for large-scale RNAi or compound screenings and candidate validation.
Main Results:
- Demonstrated the utility of high-content analysis for detailed SASP characterization.
- Enabled rapid identification and validation of potential OIS regulators.
- Showcased the system's capability for screening siRNA to modulate SASP expression.
Conclusions:
- Widefield High-Content Analysis is a powerful tool for studying senescence and SASP.
- This approach facilitates the discovery of novel regulators of cellular senescence.
- The methodology supports the development of therapeutic strategies targeting SASP in aging and cancer.
