AAVrh32.33 capsid demonstrates unexpected dermal tropism regardless of immunodominant epitope

Cutaneous gene therapy has the potential to treat a wide range of skin disorders, but effective delivery remains limited by the barrier properties and immune surveillance of the skin. Here, we identify AAVrh32.33 as a potent vector for targeting dermal stromal compartments. Following systemic administration in mice, AAVrh32.33 mediated robust and durable transgene expression, with preferential targeting of dermal fibroblasts and hair follicle bulge cells. Expression peaked at 1 month and persisted for up to 2 years, highlighting its suitability for chronic conditions. To reduce immunogenicity, a dominant CD8⁺ T cell epitope was disrupted, generating the IDPΔ variant. This modification attenuated peptide-specific T cell responses while preserving stromal transduction. In human skin explants, IDPΔ achieved high levels of gene expression, primarily in dermal fibroblasts and precursors, confirming translational relevance. Finally, vectors encoding CCL17, CCL20, and CCL22 demonstrated localized targeted therapeutic gene delivery in both healthy and inflamed skin, underscoring the feasibility of using this platform to reshape local immune responses. Together, these findings establish AAVrh32.33 and IDPΔ as promising platforms for durable cutaneous gene therapy, with direct applications in diseases such as vitiligo, where long-term modulation of the dermal microenvironment is essential.