A genetically encoded system to purify senescent cells.

Hourieh Tousian, Rachael A Protzman, Timothy J Sargeant, Julian M Carosi

Lysosomal Health in Ageing, South Australian Health and Medical Research Institute (SAHMRI), Adelaide, SA, 5000, Australia; School of Pharmacy and Biomedical Science, College of Health, Adelaide University, Adelaide, SA, 5000, Australia. · pubmed

Paper

Abstract

Cellular senescence is a state of irreversible cell-cycle arrest that prevents cancer but promotes aging and inflammation. Conflicting molecular profiles of senescence may arise from contamination by neighboring proliferating cells. Here, we developed SenLect (senescence selection), a genetically encoded system for purifying senescent cells in culture. SenLect expresses mCherry-2A-PuroR under the senescence-activated miR-146a promoter, providing senescent cells a selective advantage over proliferating cells during puromycin treatment. We validated SenLect in primary and immortal cells undergoing replicative senescence or various modes of stress-induced senescence. Selected cells were enriched for cell-cycle arrest, increased cell size, and lysosomal β-galactosidase activity. Removing proliferating cells revealed proteome rewiring during senescence induced by cyclin-dependent kinase 4/6 inhibition, improving detection of senescence-associated secretory phenotype (SASP) signatures, metabolic shifts, and organelle remodeling. SenLect provides a reliable method for isolating senescent cells for downstream applications.

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Research summary

The paper demonstrates that a genetically encoded system (SenLect) can selectively purify senescent cells from mixed populations, revealing previously obscured proteomic and metabolic changes associated with senescence. This is relevant to longevity research because cellular senescence is a fundamental hallmark of aging, and the ability to isolate these cells without contamination from proliferating neighbors is critical for accurately defining the senescence-associated secretory phenotype (SASP) and developing effective senolytic or senomorphic therapies.

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