Rapid and synchronous chemical induction of replicative-like senescence via a small molecule inhibitor [microC]

Cellular senescence is now acknowledged as a key contributor to organismal ageing and late-life disease. Although popular, the study of senescence in vitro can be complicated by the prolonged and asynchronous timing of cells committing to it and its paracrine effects. To address these issues, we repurposed the small molecule inhibitor inflachromene (ICM) to induce senescence to human primary cells. Within six days of treatment with ICM, senescence hallmarks, including the nuclear eviction of HMGB1 and -B2, are uniformly induced across the cell population. By comparing various high throughput datasets from ICM-induced and replicative senescence, we uncovered significant similarities between the two states. Notably, ICM suppresses the proinflammatory secretome associated with senescence, thus alleviating most paracrine effects. In summary, ICM rapidly and synchronously induces a senescence-like phenotype that allows us to study its core regulatory program without any confounding heterogeneity. Micro-C is applied to 4 low passage IMR90 wild type samples and 4 low passage ICM-treated samples. RNA-seq and Ribo-Seq are applied to 4 low passage IMR90 wild type samples, 4 high passage IMR90 senescent samples and to 2 low passage ICM-treated samples. scRNA-seq is applied to 1 low passage IMR90 wild type samples, 1 high passage IMR90 senescent sample and to 1 low passage ICM-treated sample. FactoryRNA-seq is applied to 2 low passage IMR90 wild type samples, 2 low passage 3 days ICM-treated samples, 2 low passage 6 days ICM-treated samples and 2 low passage metaICM-treated samples. Cut&tag for CTCF and SMC1 are applied to 1 low passage IMR90 wild type sample, 1 high passage IMR90 senescent sample and to 1 low passage ICM-treated sample.