Localized and Sustained Delivery of Silencing RNA from Macroscopic Biopolymer Hydrogels

The ability to silence the expression of specific genes at a particular location of the body would provide a powerful new therapeutic too[ for treatment of diseases such as cancer or for use in regenerative medicine. RNA interference (RNAi) is a gene silencing mechanism where specific mRNA molecules that are complementary to short interfering RNA (siRNA) are degraded, thus inhibiting gene expression at the post-transcriptional level. However, the use of siRNA has not yet realized its full clinical potential due to degradation in vivo, the difficulty retaining siRNA at the site of interest, and the relatively short-term effect it has on rapidly dividing cells. In this work a new paradigm is presented that will allow for the localized delivery of siRNA that is controlled and sustained over time, thus allowing cells at the site of interest to be directly exposed to a gradual release of bioactive siRNA. To accomplish this, three different types of macroscopic, degradable biomaterial hydrogel. scaffolds were employed: calcium crosslinked alginate, photocrosslinked alginate, and collagen. Differing rates of release from these hydrogels were achieved, and the ability of the released siRNA to knock down the expression of GFP in cells that constitutively express this protein was shown. Furthermore, the ability to encapsulate cells within these materials and achieve sustained gene silencing of these incorporated cells was demonstrated. These biopolymer hydrogels are injectable and, therefore, can be delivered in a minimally invasive manner, and they can serve as delivery vehicles for both siRNA and transplanted cell populations.