Reprogramming Factors Activate a Non-Canonical Oxidative Resilience Pathway That Can Rejuvenate RPEs and Restore Vision

Lu, Y. R.; Cameron, J. C.; Hu, Y.; Shen, H.; Shirahama, S.; Tyshkovskiy, A.; Chen, Z.; Ai, J.; Zhu, D. Y.; Karg, M. M.; Chew, L. A.; Bell, G. W.; Jena, S. G.; He, Y.; Seifert, P.; Shu, D. Y.; El-Brolosy, M. A.; Lou, Q.; Zhang, B.; Puszynska, A. M.; Qiu, X.; Tian, X.; Gregory-Ksander, M.; Gladyshev, V. N.; Sinclair, D. A.; Saint-Geniez, M.; Buenrostro, J. D.; Rickman, C. B.; Ksander, B. R.; Weissman, J. S.

doi:10.1101/2025.08.30.673239

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bioRxiv · 10.1101/2025.08.30.673239

Reprogramming Factors Activate a Non-Canonical Oxidative Resilience Pathway That Can Rejuvenate RPEs and Restore Vision

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Abstract

Oct4, Sox2, and Klf4 (OSK) Yamanaka factors induce pluripotency and reverse age-related epigenetic changes, yet the mechanisms by which they promote rejuvenation remain poorly explored. Oxidative stress contributes to CNS aging and retinal pigmented epithelium (RPE) degeneration in age-related macular degeneration. We find that OSK expression in RPE restores retinal structure and visual function in aged mice and promotes oxidative resilience through a non-canonical, Tet2-independent pathway. Integrative functional genomics identifies GSTA4, a detoxifying enzyme that clears the lipid peroxidation byproduct 4-HNE, as a necessary and sufficient OSK effector. Dynamic GSTA4 regulation by OSK recapitulates a stem cell derived stress resilience program. GSTA4 overexpression alone enhances mitochondrial resilience, rejuvenates the aged RPE transcriptome, and reverses visual decline. GSTA4 is consistently upregulated across diverse lifespan-extending interventions suggesting a broader pro-longevity role. These findings uncover a previously unrecognized protective axis driven by Yamanaka factors that circumvents reprogramming, providing therapeutic insights for age-related diseases. HIGHLIGHTSO_LIOSK-GSTA4 provides a dynamic, Tet2-independent stress-resilience axis. C_LIO_LIFunctional genomics pinpoints GSTA4 as a direct downstream effector activated by OSK. C_LIO_LIRPE aging involves progressive accumulation of 4-HNE that can be detoxified by GSTA4. C_LIO_LIEnhancing GSTA4 rejuvenates RPE cells, restores vision and is associated with lifespan-extending interventions. C_LI

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Lu, Y. R., Cameron, J. C., Hu, Y., Shen, H., Shirahama, S., Tyshkovskiy, A., Chen, Z., Ai, J., Zhu, D. Y., Karg, M. M., Chew, L. A., Bell, G. W., Jena, S. G., He, Y., Seifert, P., Shu, D. Y., El-Brolosy, M. A., Lou, Q., Zhang, B., Puszynska, A. M., Qiu, X., Tian, X., Gregory-Ksander, M., Gladyshev, V. N., Sinclair, D. A., Saint-Geniez, M., Buenrostro, J. D., Rickman, C. B., Ksander, B. R., Weissman, J. S.. 2025-09-01. Reprogramming Factors Activate a Non-Canonical Oxidative Resilience Pathway That Can Rejuvenate RPEs and Restore Vision. https://doi.org/10.1101/2025.08.30.673239

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