ArticleGeroScience2026
Quantitative Cellular AGing Evaluation system (qCAGEs): a dual-parameter platform for high-throughput senescence screening.
Article in GeroScience, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Accurate identification and quantification of senescence-modulating compounds require screening platforms that can distinguish between phenotypically distinct drug response profiles. Conventional approaches relying on single-parameter measurements-either cell viability or senescence markers alone-cannot differentiate senolytic-like or anti-aging-like response profiles from non-specific cytotoxicity or proliferative effects. Here, we present the quantitative Cellular AGing Evaluation system (qCAGEs), a dual-parameter high-throughput screening platform that simultaneously measures aging-associated α-L-fucosidase activity (4ME) and cell viability. By establishing the Cell count-Aging activity Reference Line (CARL), which defines the expected relationship between cell number and 4ME activity under baseline conditions, qCAGEs enables the systematic phenotypic classification of drug responses into four operational phenotypic response classes-anti-aging-like, pro-aging-like, senolytic-like, and cytotoxic-defined by their position relative to baseline cell abundance and the reference 4ME-count relationship. The quantitative Cellular Aging Index (qCAI), calculated as the perpendicular distance from each response coordinate to the CARL, provides a continuous measure of the magnitude of the drug effect within each classification category. Using human dermal fibroblast senescence models, we validated that the 4ME/cell count ratio robustly discriminated senescent from young cells (> sixfold difference), with assay precision meeting the high-throughput screening quality criteria (coefficient of variation (CV) < 15%). The platform was successfully adapted to three-dimensional culture formats using 384-well micropillar plates with ATP-based viability quantification, demonstrating strong concordance with two-dimensional results (cosine similarity > 0.5 for > 75% of compounds). The qCAGEs framework addresses a critical unmet need in senescence research by providing a standardized, quantitative approach for evaluating drug-induced changes in cellular aging status, with broad applications in senolytic and anti-aging therapeutic development.
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