A ligand/receptor trafficking clock governs self-renewal and abscission dynamics in pluripotent stem cells

Libetti, D.; Combemorel, N.; Fraimbault, E.; Lantelme, M.; Gaume, X.; Zakiev, E.; Tapponnier, Y.; Huyghe, A.; Furlan, G.; Coudurier, M.; Claret, L.; rognard, c.; Akguel, S.; Hernandez-vargas, H.; Gadot, N.; Forest, F.; Koch, M.; Aksoy, I.; Allegre, N.; Chazaud, C.; Ramalho-santos, M.; Savatier, P.; Bachelard-cascales, E.; Lavial, F.

doi:10.64898/2025.12.22.695902

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bioRxiv · 10.64898/2025.12.22.695902

A ligand/receptor trafficking clock governs self-renewal and abscission dynamics in pluripotent stem cells

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Abstract

Summary/AbstractHow extracellular cues are temporally integrated to regulate self-renewal and differentiation propensities across the cell cycle remains largely unresolved. We identify a ligand/receptor trafficking clock in rodent and human pluripotent stem cells (PSCs) in which the cyclic turnover of Netrin-1 and its receptors Neo1 and Unc5b (NNU) governs self-renewal capacity and abscission dynamics. In G1, NNU complexes undergo Clathrin-mediated internalization and lysosomal degradation, a process required for timely post-mitotic bridge abscission. At later stages of the cycle, NNU activate Src within early endosomes, inducing a genome-wide redistribution of the transcriptional co-activator Yap1. This reshapes gene regulatory networks by activating stemness- and ectoderm-associated transcriptional programs enriched for Sox2/Nanog binding and by repressing mesodermal- and cell cycle-related targets enriched for Sox2 and Tcf3. Functionally, recombinant Netrin-1 reduces functional heterogeneity and enhances clonogenicity in G1, uncovering a tractable strategy to canalize stem cell behavior. Collectively, our results reveal cell cycle-dependent ligand/receptor trafficking as a temporal clock that directly links membrane dynamics to epigenetic regulation and stem cell fate, opening new avenues for regenerative medicine.

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Libetti, D., Combemorel, N., Fraimbault, E., Lantelme, M., Gaume, X., Zakiev, E., Tapponnier, Y., Huyghe, A., Furlan, G., Coudurier, M., Claret, L., rognard, c., Akguel, S., Hernandez-vargas, H., Gadot, N., Forest, F., Koch, M., Aksoy, I., Allegre, N., Chazaud, C., Ramalho-santos, M., Savatier, P., Bachelard-cascales, E., Lavial, F.. 2025-12-23. A ligand/receptor trafficking clock governs self-renewal and abscission dynamics in pluripotent stem cells. https://doi.org/10.64898/2025.12.22.695902

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