Receptor-Guided AAV Tropism Engineering via MATCH.

Area of Science:

• Biotechnology
• Molecular Biology
• Gene Therapy

Background:

• Controlling viral tropism is crucial for effective gene delivery.
• Adeno-associated viruses (AAVs) are promising gene therapy vectors but lack precise targeting.
• Current methods for modifying AAV tropism are limited.

Purpose of the Study:

• To develop a modular method for programmable retargeting of AAVs.
• To enable receptor-guided modification of AAV tropism for enhanced gene delivery.
• To create a scalable and versatile platform for custom AAV vector design.

Main Methods:

• Developed MATCH (Modulation of AAV Tropism through Conjugation to Homing proteins), a site-specific covalent protein conjugation technique.
• Incorporated SpyTag peptide into AAV capsid loops for one-step attachment of SpyCatcher-linked targeting proteins.
• Utilized mosaic AAV-DJ and AAV9 capsids for tunable ligand display and efficient assembly.
• Established a streamlined one-pot "Mix-and-MATCH" production strategy for coexpression of capsid and targeting ligands.

Main Results:

• MATCH-AAVs conjugated to anti-CD3 antibodies efficiently transduced human T cells in vitro (up to 58% of PBMCs).
• TfR1-targeted MATCH-AAV9 vectors showed up to 88-fold increased brain expression in mice.
• Human TfR1-targeted vectors demonstrated robust, receptor-dependent transduction in vitro and in humanized mouse models.
• TfR1-targeted vectors facilitated widespread parenchymal transduction, indicating blood-brain barrier crossing.

Conclusions:

• MATCH provides a versatile synthetic-biology toolkit for rational AAV tropism engineering.
• This method enables programmable, receptor-guided retargeting of AAVs for research and therapeutic applications.
• The "Mix-and-MATCH" strategy offers a scalable route to custom AAV vector production.