A Versatile Adeno-Associated Viral Vector Cross-Linking Platform Capable of Tuning Cellular Tropisms and Simultaneously Inducing Solid-Phase Gene Delivery.

Abstract

Developing gene carriers with improved affinities for target cells and the simultaneous diversification of their delivery modes will be pivotal for upgrading gene therapy technologies. In this study, a simple and versatile adeno-associated virus (AAV) conjugation platform using the cross-linker 3,3'-dithiobis(sulfosuccinimidyl propionate) (DTSSP) is proposed. Depending on the quantity of the DTSSP molecules, the AAV-DTSSP complexes could either be linked with the relevant biomolecules for altering cellular tropisms or further form a self-assembled AAV-DTSSP pellet capable of mimicking a polymeric gene delivery system. At lower quantities of DTSSP, the AAV-DTSSP complexes were conjugated with aminated l-fucose molecules, whose levels are typically upregulated in pancreatic cancer cells, resulting in enhanced gene delivery efficiencies in pancreatic cancer cells. At higher concentrations of DTSSP, visible solid forms of the AAV-DTSSP pellets were formed, and the AAV pellets demonstrated the capability to induce a localized, sustained gene expression pattern comparable to that of conventional biomaterial-based approaches. Thus, a multipurpose AAV cross-linking platform, which can enable AAV vector systems that are capable of altering cellular tropisms and simultaneously inducing solid-phase delivery, will provide crucial insights into vector design for further upgrading of gene delivery technologies.