ABSTRACT
With nucleic acid delivery widely brought into the foray through mRNA vaccine formulations, its combination with gene editing technology poses a novel therapeutic approach for inheritable monogenic diseases. By delivering Cas9 mRNA with appropriate guide RNAs (gRNA) and template DNA, nucleic acid therapies could address monogenic diseases at the genetic level. Poly(amine-co-ester), or PACE, is a biodegradable, biocompatible, and tunable polymer capable of carrying nucleic acid cargo into cells. Previously, we have optimized and demonstrated efficacy of the PACE platform for lung transfection, specifically through intratracheal (IT) or intranasal (IN) administration of PACE-mRNA polyplexes for vaccine purposes. However, the potential of PACE to deliver a combination of nucleic acids and facilitate gene editing has not yet been fully realized. Furthermore, enhancing delivery by attaching targeting moieties, which would endow higher accumulation in cells of interest while sparing off-target cells, would offer a new functionality that lends versatility to this potent platform.
Our central hypothesis is that PACE polyplex transfection by IT administration can be enhanced with targeting moieties and, in combination with gene editing technology, can provide a novel therapeutic approach for monogenic inheritable disease in the lung, specifically cystic fibrosis (CF). We aim to characterize which lung cell populations are susceptible to transfection by PACE and develop a targeted PACE platform that will lead to optimal transfection of cells of interest. We also seek to apply PACE to a monogenic disease model, specifically CF, thus demonstrating the potential of PACE as a customizable delivery platform within the field of gene therapy.