Ageing impairs the ability of neural stem cells (NSCs) to transition from quiescence to proliferation in the adult mammalian brain. Functional decline

CRISPR–Cas9 screens reveal regulators of ageing in neural stem cells

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2024-10-04 01:30:03

Ageing impairs the ability of neural stem cells (NSCs) to transition from quiescence to proliferation in the adult mammalian brain. Functional decline of NSCs results in the decreased production of new neurons and defective regeneration following injury during ageing1,2,3,4. Several genetic interventions have been found to ameliorate old brain function5,6,7,8, but systematic functional testing of genes in old NSCs—and more generally in old cells—has not been done. Here we develop in vitro and in vivo high-throughput CRISPR–Cas9 screening platforms to systematically uncover gene knockouts that boost NSC activation in old mice. Our genome-wide screens in primary cultures of young and old NSCs uncovered more than 300 gene knockouts that specifically restore the activation of old NSCs. The top gene knockouts are involved in cilium organization and glucose import. We also establish a scalable CRISPR–Cas9 screening platform in vivo, which identified 24 gene knockouts that boost NSC activation and the production of new neurons in old brains. Notably, the knockout of Slc2a4, which encodes the GLUT4 glucose transporter, is a top intervention that improves the function of old NSCs. Glucose uptake increases in NSCs during ageing, and transient glucose starvation restores the ability of old NSCs to activate. Thus, an increase in glucose uptake may contribute to the decline in NSC activation with age. Our work provides scalable platforms to systematically identify genetic interventions that boost the function of old NSCs, including in vivo, with important implications for countering regenerative decline during ageing.

The adult mammalian brain contains several NSC regions that give rise to newborn neurons and can repair tissue damaged by stroke or brain injuries1,4,9,10,11,12,13,14. The most active NSC niche is located in the subventricular zone (SVZ) that lines the lateral ventricles of the brain1,2,4,10,13,14,15,16,17,18. NSCs from the SVZ region can generate thousands of newborn neurons each day in a young adult mouse10. The SVZ region comprises a pool of quiescent NSCs (qNSCs) that can give rise to activated (proliferating) NSCs (aNSCs), which in turn generate more committed progenitors that migrate out of the niche towards the olfactory bulb, where they differentiate into neurons. The ability of NSCs to activate and form newborn neurons is severely impaired in the ageing brain, and this can contribute to deficits in sensory and cognitive function1,2,3,15,19,20,21,22,23.