Spontaneously slow-cycling subpopulations of human cells originate from activation of stress-response pathways

Slow-cycling subpopulations exist in bacteria, yeast, and mammalian systems. In the case of cancer, slow-cycling subpopulations have been proposed to give rise to drug resistance. However, the origin of slow-cycling human cells is poorly studied, in large part due to lack of markers to identify these rare cells. Slow-cycling cells pass through a noncycling period marked by low CDK2 activity and high p21 levels. Here, we use this knowledge to isolate these naturally slow-cycling cells from a heterogeneous population and perform RNA sequencing to delineate the transcriptome underlying the slow-cycling state. We show that cellular stress responses—the p53 transcriptional response and the integrated stress response (ISR)—are the most salient causes of spontaneous entry into the slow-cycling state. Finally, we show that cells’ ability to enter the slow-cycling state enhances their survival in stressful conditions. Thus, the slow-cycling state is hardwired to stress responses to promote cellular survival in unpredictable environments.
Single-cell time-lapse imaging and transcriptomic analysis reveal why some human cells proliferate more slowly than the majority of the population, with cellular stress responses triggering entry into a slow-cycling state to promote cellular survival in unpredictable environments.
Author summary Even within a genetically identical population, some cells proliferate more slowly than others. Slow-cycling cells have been implicated in resistance to antibiotics, antifungals, and cancer therapies, yet the origin of the slow-cycling state remains poorly understood. Here, we isolate a naturally slow-cycling subpopulation of human cells and find that the slow-cycling state is induced by moderate activation of stress responses. We further show that the ability to enter this slow-cycling state protects cells from further stress, consistent with its association with drug resistance. We propose that the existence of the slow-cycling state thereby promotes long-term survival of populations that occasionally experience mildly stressful environments.