Gene transfer to hemophilia A mice via oral delivery of FVIII–chitosan nanoparticles

Abstract

Effective oral delivery of a non-viral gene carrier would represent a novel and attractive strategy for therapeutic gene transfer. To evaluate the potential of this approach, we studied the oral gene delivery efficacy of DNA polyplexes composed of chitosan and Factor VIII DNA. Transgene DNA was detected in both local and systemic tissues following oral administration of the chitosan nanoparticles to hemophilia A mice. Functional factor VIII protein was detected in plasma by chromogenic and thrombin generation assays, reaching a peak level of 2-4% FVIII at day 22 after delivery. In addition, a bleeding challenge one month after DNA administration resulted in phenotypic correction in 13/20 mice given 250-600 μg of FVIII DNA in chitosan nanoparticles, compared to 1/13 mice given naked FVIII DNA and 0/6 untreated mice. While further optimization would be required to render this type of delivery system practical for hemophilia A gene therapy, the findings suggest the feasibility of oral, non-viral delivery for gene medicine applications.

Development and Evaluation of Chitosan-Coated Liposomes for Oral DNA Vaccine: The Improvement of Peyer’s Patch Targeting Using a Polyplex-Loaded Liposomes

“…The uses of chitosan as cationic complexing agent and as coating polymer have been extensively studied for oral drug delivery systems (9,17,34). As coating agent, the chitosan polymer decreased the chance of lipid vesicles to be attacked by enzymes.…”

Mechanistic studies on the uptake and intracellular trafficking of novel cyclodextrin transfection complexes by intestinal epithelial cells

“…Local expression of therapeutic proteins may have therapeutic benefit in the treatment of inflammatory bowel disease, intestinal cancer, and the intestinal symptoms of cystic fibrosis (O' Neill et al 2011) (Page and Cudmore 2001). Furthermore, the intestine can potentially provide a route of entry into the bloodstream for locally expressed therapeutic proteins such as Factor VIII in haemophilia patients (Bowman et al 2008), or indeed for gene therapies to distant disease sites.…”

Oral gene application using chitosan-DNA nanoparticles induces transferable tolerance

“…Chitosan-DNA NP are of increasing interest as a nonviral gene carrier system (5,6,27). Using chitosan-DNA NP for oral application, exploiting oral tolerance mechanisms, would allow the development of DNA-based tolerogenic vaccines.…”

“…Chitosan has been complexed with plasmid DNA, forming chitosan-DNA nanoparticles (NP), which are stable during the gastrointestinal passage and will be phagocytized in the gut, resulting in gene expression (2). It was shown that feeding of factor VIII-encoding chitosan-DNA NP to hemophilia A mice resulted in increased factor VIII plasma levels (6,15) and that oral application of erythropoietin-encoding chitosan-DNA NP led to a significant increase of hematocrit levels (8). In rodent models of diabetes, chitosan-DNA NP encoding insulin or glucagon-like peptide 1 were able to decrease blood glucose concentrations (23,31,32).…”