Netrin-1 Acts as a Guardian of Naive Pluripotency in Human Embryonic Stem Cells

De Neufville, A.; Masfaraud, E.; Alfeghaly, C.; Stoeckl, J. B.; Doerflinger, N.; Marcy, G.; Rognard, C.; OSTEIL, P.; Lantelme, M.; Lavial, F.; Chazaud, C.; Frohlich, T.; Savatier, P.; Aksoy, I.

doi:10.64898/2026.07.16.738622

bioRxiv · 10.64898/2026.07.16.738622

Netrin-1 Acts as a Guardian of Naive Pluripotency in Human Embryonic Stem Cells

De Neufville, A.·Masfaraud, E.·Alfeghaly, C.·Stoeckl, J. B.·Doerflinger, N.·Marcy, G.·Rognard, C.·OSTEIL, P.·Lantelme, M.·Lavial, F.·Chazaud, C.·Frohlich, T.·Savatier, P.·Aksoy, I.

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Abstract

We investigated the role of Netrin-1 (NTN1) in human naive pluripotency using complementary loss- and gain-of-function approaches. In primate embryos and human embryonic stem cells (hESCs), Netrin-1 expression is associated with the naive pluripotent state. Disruption of NTN1 had no detectable effect on hESCs maintained on murine embryonic fibroblasts. However, under sub-optimal culture conditions, NTN1-knockout cells exhibited compromised naive pluripotency, which was rescued with feeder cells overexpressing Netrin-1. Netrin-1 overexpression in hESCs accelerated acquisition of the naive state and markedly increased resistance to differentiation. These effects were accompanied by extensive epigenetic remodeling, including H3K27ac and H2K27me3. Proteomic and phospho-proteomic analyses further revealed rapid Netrin-1-dependent alterations in pathways controlling cell adhesion, signaling, and chromatin regulation. Together, these findings extend the role of Netrin-1 beyond its established functions and identify it as a coordinator of extracellular cues, intracellular signaling, and nuclear regulatory mechanisms that support human naive pluripotency.

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De Neufville, A., Masfaraud, E., Alfeghaly, C., Stoeckl, J. B., Doerflinger, N., Marcy, G., Rognard, C., OSTEIL, P., Lantelme, M., Lavial, F., Chazaud, C., Frohlich, T., Savatier, P., Aksoy, I.. 2026-07-16. Netrin-1 Acts as a Guardian of Naive Pluripotency in Human Embryonic Stem Cells. https://doi.org/10.64898/2026.07.16.738622

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