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bioRxiv · 10.1101/2025.06.05.657370

A single factor for safer cellular rejuvenation

Abstract

Ageing is a key driver of the major diseases afflicting the modern world. Slowing or reversing the ageing process would therefore drive significant and broad benefits to human health. Previously, the Yamanaka factors (OCT4, SOX2, KLF4, with or without c-MYC: OSK(M)) have been shown to rejuvenate cells based on accurate predictors of age known as epigenetic clocks. Unfortunately, OSK(M) induces dangerous pluripotency pathways, making it unsuitable for therapeutic use. To overcome this therapeutic barrier, we screened for novel factors by optimising directly for age reversal rather than for pluripotency. We trained a transcriptomic ageing clock, unhindered by the low throughput of bulk DNA methylation assays, to enable a screen of unprecedented scale and granularity. Our platform identified SB000, the first single gene intervention to rejuvenate cells from multiple germ layers with efficacy rivalling the Yamanaka factors. Cells rejuvenated by SB000 retain their somatic identity, without evidence of pluripotency or loss of function. These results reveal that decoupling pluripotency from cell rejuvenation does not remove the ability to rejuvenate multiple cell types. This discovery paves the way for cell rejuvenation therapeutics that can be broadly applied across age-driven diseases. HighlightsO_LISB000 drives multi-omic rejuvenation in human fibroblasts, as evidenced by substantial reversal of numerous epigenetic clocks, lowered single-cell transcriptomic age, and decreased senescence-associated gene expression. C_LIO_LIIn contrast to OSK(M), SB000 treatment maintains transcriptomic and functional measures of fibroblast identity without the activation of pluripotency. C_LIO_LISB000 rejuvenation generalises to keratinocytes, cells from another germ layer, with potency matching or surpassing OSK(M). C_LI

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BibTeXRIS

de Lima Camillo, L. P., Gam, R., Maskalenka, K., LeBlanc, F. J., Urrutia, G. A., Mejia, G. M., Miller, H. E., Wardlaw, C. P., Pickles, A., Everton, L., Zaksauskaite, R., Khan, R. B., Welsh, A., Gambo, S., Gallardo, S., Oliynyk, Z., Varankar, S. S., Epstein, A. E., Bendall, A., Mowatt, J., Ives, D., Swain, B. M.. 2025-06-06. A single factor for safer cellular rejuvenation. https://doi.org/10.1101/2025.06.05.657370

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