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

Area of Science:

• Nanotechnology
• Gene Therapy
• Immunology

Background:

• Recombinant adeno-associated viruses (rAAVs) show potential for orthopedic diseases.
• Conventional AAV vectors face challenges including immunogenicity and low potency, requiring high doses.
• High doses increase costs and safety concerns for AAV-based therapies.

Purpose of the Study:

• To investigate the use of Pluronic F-127 (PF127) micelles encapsulating rapamycin (PF127-RAPA) to enhance rAAV efficacy.
• To determine the mechanisms by which PF127-RAPA improves AAV transduction and reduces immunogenicity.
• To evaluate the therapeutic potential of PF127-RAPA in preclinical models of osteoimmunological disorders.

Main Methods:

• Co-administration of AAV vectors with PF127-RAPA.
• Analysis of AAV nuclear entry and cytoplasmic accumulation.
• Assessment of MHC class I presentation of capsid proteins.
• Evaluation of therapeutic efficacy in mouse models of postmenopausal osteoporosis and rheumatoid arthritis.

Main Results:

• PF127-RAPA significantly enhanced AAV transduction efficiency and reduced immunogenicity.
• PF127-RAPA modulated nuclear membrane properties, increasing passive transport via nuclear pore complexes (NPCs) for enhanced AAV nuclear entry.
• PF127-RAPA decreased cytoplasmic AAV accumulation, minimizing immune presentation of capsid proteins.
• A 75% dosage reduction of AAV6-SEMA3A was achieved with PF127-RAPA while maintaining potent efficacy in disease models.

Conclusions:

• PF127-RAPA is a promising nanotechnology approach to improve AAV-based gene therapies for osteoimmunological disorders.
• This method enhances AAV delivery and reduces immune responses, allowing for lower, safer, and more cost-effective therapeutic doses.
• The findings support the clinical translatability of PF127-RAPA for enhancing AAV therapies.