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The PICLS high-throughput screening method for agents extending cellular longevity identifies 2,5-anhydro-D-mannitol
Mohammad Alfatah1, Frank Eisenhaber2,3,4
1Bioinformatics Institute (BII), Singapore, 138671, A*STAR, Singapore. alfatahm@bii.a-star.edu.sg.
Abstract:
Although aging is the biggest risk factor for human chronic (cancer, diabetic, cardiovascular, and neurodegenerative) diseases, few interventions are known besides caloric restriction and a small number of drugs (with substantial side effects) that directly address aging. Thus, there is an urgent need for new options that can generally delay aging processes and prevent age-related diseases. Cellular aging is at the basis of aging processes. Chronological lifespan (CLS) of yeast Saccharomyces cerevisiae is the well-established model system for investigating the interventions of human post-mitotic cellular aging. CLS is defined as the number of days cells remain viable in a stationary phase. We developed a new, cheap, and fast quantitative method for measuring CLS in cell cultures incubated together with various chemical agents and controls on 96-well plates. Our PICLS protocol with (1) the use of propidium iodide for fluorescent-based cell survival reading in a microplate reader and (2) total cell count measurement via OD600nm absorption from the same plate provides real high-throughput capacity. Depending on logistics, large numbers of plates can be processed in parallel so that the screening of thousands of compounds becomes feasible in a short time. The method was validated by measuring the effect of rapamycin and calorie restriction on yeast CLS. We utilized this approach for chemical agent screening. We discovered the anti-aging/geroprotective potential of 2,5-anhydro-D-mannitol (2,5-AM) and suggest its usage individually or in combination with other anti-aging interventions.
Insights
Researchers developed a fast, high-throughput method to screen for aging interventions. They discovered 2,5-anhydro-D-mannitol (2,5-AM) shows anti-aging potential in yeast, a model for cellular aging.
Area of Science:
- Gerontology and cellular aging research.
- Yeast Saccharomyces cerevisiae as a model for aging studies.
Background:
- Aging is the primary risk factor for chronic diseases, yet effective interventions are limited.
- Existing anti-aging strategies like caloric restriction have drawbacks, necessitating new therapeutic options.
- Cellular aging underpins organismal aging, making it a key target for interventions.
Purpose of the Study:
- To develop a novel, high-throughput method for measuring chronological lifespan (CLS) in yeast.
- To screen chemical compounds for their potential to delay aging processes and extend lifespan.
- To identify new geroprotective agents for potential use in human aging.
Main Methods:
- Developed the Propium Iodide-based Cell Lifespan (PICLS) assay using 96-well plates for quantitative CLS measurement.
- Integrated fluorescent cell survival reading (propidium iodide) and total cell count (OD600nm) in a single assay.
- Validated the PICLS method using known aging modulators like rapamycin and caloric restriction.
Main Results:
- The PICLS assay enables rapid, cost-effective screening of thousands of compounds for anti-aging effects.
- Validated the method's reliability and high-throughput capacity for aging research.
- Identified 2,5-anhydro-D-mannitol (2,5-AM) as a novel compound with significant anti-aging and geroprotective potential in yeast.
Conclusions:
- The PICLS protocol is a powerful tool for accelerating the discovery of anti-aging interventions.
- 2,5-anhydro-D-mannitol (2,5-AM) demonstrates promise as a geroprotective agent.
- This discovery opens avenues for developing new strategies to combat age-related diseases.
