Digoxigenin modification of adenovirus to spatially control gene delivery from chitosan surfaces

Abstract

To spatially control the delivery of multiple viral vectors from biomaterial scaffolds, digoxigenin (DIG) was conjugated to adenoviral capsid proteins as an antigenic determinant for antibody immobilization. The infectivity, toxicity, specificity and immobilization stability of DIG-modified adenovirus were examined to investigate the feasibility and effectiveness of this viral surface modification. Anti-DIG antibody conjugated on chitosan surfaces was able to immobilize DIGmodified adenovirus and could be stably bound on the material for at least two weeks, yet the modification was mild enough that viral infectivity was maintained. To immobilize two different adenoviruses, wax masking was applied to conjugate anti-DIG and anti-adenovirus antibodies in two discrete regions of a chitosan film, respectively. The distribution of these two viral vectors expressing different reporter genes was examined after cell culture. Fluorescent protein expression from transduced cells illustrated that the infection distribution could be controlled: one gene was delivered to the entire region of the biomaterial, and another was only delivered to defined regions. Compared to three other cardiac glycosides, ATPase inhibition was undetectable when DIG was conjugated on the adenovirus, suggesting that the method may be safe for in vivo application. This dual viral vector delivery system should be capable of generating distinct interfaces between cell signaling viruses to control tissue regeneration from a range of different biomaterials.

Cardiac Glycoside Glucoevatromonoside Induces Cancer Type-Specific Cell Death

“…We previously documented the inhibitory potential of GEV on Na + /K + -ATPase activity (Bertol et al, 2011 ) by an in vitro assay (Hu et al, 2009 ). To validate the interaction mode of GEV and its putative pharmacological target, the Na+/K+ ATPase, we implemented docking simulations using Autodock Vina software (Trott and Olson, 2010 ) (Figure 1B ).…”

Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

“…An important feature of the current study includes the use of self-inactivating lentiviral vectors in place of γ-retrovirus (43), nonintegrating viral vectors (16,(40)(41)(42)55), or naked plasmid DNA (15). Although considerable progress has been made with regard to spatial control of gene delivery to cells, transient gene expression and poor transfection/transduction efficiencies may not allow sufficient control of cell phenotype and tissue development.…”

A Synthetic Gene Circuit for Self-Regulating Delivery of Biologic Drugs in Engineered Tissues

“…In previous approaches, gene therapy for the treatment of OA has been performed with plasmid DNA, 46 retrovirus, 47 lentivirus, [48][49][50] and most commonly non-integrating viral vectors such as adeno-associated virus. 37,[51][52][53][54] Lentiviral delivery is advantageous for this system due to its ability to stably integrate into the genome of dividing and non-dividing cells for long-term gene expression, its larger packaging capacity, low immunogenicity, and low cytotoxicity. 55,56 The vector used in this study is self-inactivating and, therefore, replication-defective, overcoming safety concerns previously associated with viral gene therapy.…”