A TFEB-TGFß axis systemically regulates diapause, stem cell resilience and protects against senescence.

Diapause is a long-lived state of resilience that allows organisms to outlast adversity. C. elegans can endure months in a fasting-induced adult reproductive diapause (ARD) and, upon refeeding, regenerate and reproduce. Here, we find that mutants of ARD master regulator hlh-30/TFEB arrest in a novel senescence-like state during ARD and refeeding, in which germline stem cells are characterized by DNA damage, nucleolar expansion, cell cycle arrest, and mitochondrial dysfunction, alongside dysregulated immune and growth metabolic signatures, elevated senescence-associated ß-galactosidase and premature aging at the organismal level. Forward genetic screens reveal a TFEB-TGFß signaling axis that systemically controls diapause, stem cell longevity, and senescence, aligning nutrient supply to proper metabolism and growth signaling. hlh-30 mutation misaligns nutrient cues and growth signaling, triggering senescence and abrogating stem cell and organismal longevity, which can be restored by downregulating TGFß signaling. Notably, TFEB's vital role is conserved in mouse embryonic and human cancer diapause. Thus, ARD offers a powerful model to rapidly characterize diapause-related regulatory pathways and study stem cell longevity and senescence in vivo, directly relevant to mammals. Overall design: RNA-seq profiling of wildtype C. elegans, hlh-30(tm1978), hlh-30(tm1978) daf-1(m40), daf-1(m40), at 48 hours ARD and at 12 hours refeeding after 48 hours of ARD