Modulating nuclear stiffness and envelope barrier facilitates AAV nuclear entry and reduces immunogenicity

Recombinant adeno-associated viruses (rAAVs) have shown promise for orthopedic diseases. However, conventional AAV vectors face major challenges, where immune responses reduce transgene efficacy and inadequate vector potency requires higher doses, leading to increased costs and safety concerns. Here, we show that Pluronic F-127 (PF127) micelles encapsulating rapamycin (PF127-RAPA), co-administered with AAV vectors, significantly enhanced AAV transduction efficiency and reduce its immunogenicity. Mechanistically, PF127-RAPA modulates the morphology and mechanical properties of the nuclear membrane, which is associated with enhanced passive transport mediated by nuclear pore complexes (NPCs) and thereby increased AAV nuclear entry. Concurrently, PF127-RAPA reduced AAV particle accumulation in the cytoplasm, minimizing MHC class I presentation of capsid proteins and attenuating the cellular immune response. Notably, in postmenopausal osteoporosis and rheumatoid arthritis mouse models, PF127-RAPA enabled AAV6-SEMA3A to achieve a 75% reduction in dosage while maintaining potent efficacy. This work presents a clinically translatable, cost-effective nanotechnology approach to enhance AAV-based therapies for osteoimmunological disorders.