Enhancer-directed gene delivery for digit regeneration based on conserved epidermal factors.
Publication/Presentation Date
12-2-2025
Abstract
UNLABELLED: Limb loss remains a significant clinical challenge, but regenerative medicine approaches such as gene therapy offer a promising strategy to trigger endogenous regeneration programs. Optimal vector configurations and molecular targets for appendicular skeletal repair are not well defined. Here, we leveraged insights from species with a high endogenous capacity for appendage regeneration to design an enhancer-directed gene delivery platform that functions during mouse digit regeneration, a well characterized model for partial limb regeneration in mammals. Single-cell RNA sequencing of zebrafish caudal fin regeneration, combined with expression data in regenerating salamander limbs and mouse digit tips, implicated the SP family of transcription factors as conserved, epidermally-expressed mediators of appendage regrowth. Null mutants of
SIGNIFICANCE STATEMENT: Instructing regeneration of complex structures in mammals remains an unsolved problem. Gene therapy offers a compelling approach to foster endogenous regeneration by delivering therapeutic gene products to specific cells post injury. We identified a conserved regeneration-linked epidermal transcriptional program in mouse digit regeneration centered on the SP6 and SP8 transcription factors, involving inflammatory responses from osteoclasts. We engineered AAVs harboring a zebrafish tissue regeneration enhancer to direct FGF8 expression in the epidermis after amputation. This enhancer directed delivery partially rescued impaired digit regeneration in
ISSN
2692-8205
Published In/Presented At
Brown, D. A., Koll, K. K., Brush, E., Darner, G., Curtis, T., Dvergsten, T., Tran, M., Milligan, C., Wolfson, D., Gonzalez, T. J., Jeffs, S., Ehrhardt, A., Bitolas, R., Landau, M., Reitz, K., Salven, D. S., Slota-Burtt, L. A., Snee, I., Singer-Freeman, E., Bhatia, S., … Poss, K. D. (2025). Enhancer-directed gene delivery for digit regeneration based on conserved epidermal factors. bioRxiv : the preprint server for biology, 2025.12.01.691633. https://doi.org/10.64898/2025.12.01.691633
PubMedID
41573951
Department(s)
Medical Education
Document Type
Article