Platform for Designing and Delivering Tissue-Specific, Payload-Unrestricted, Redosable, Non-Viral Gene Therapy for Any Indication

PROJECT SUMMARY – Gene therapy has the potential to treat or cure thousands of rare diseases, and it holds great promise for treating or reducing the risk of numerous other conditions. However, the development of gene therapies and their clinical deployment have been plagued by several challenges including 1) the high cost of manufacturing, 2) the inability or limited ability to target specific tissues, 3) the limited size of genes that can be incorporated in a therapy, and 4) limited options for redosing given the potential for an immunogenic response to the gene delivery system. Several of these limitations stem from the use of viral vectors as gene delivery systems, but even non-viral systems like transpon-based delivery and liposomal delivery have payload size restrictions and safety concerns that have so far prevented their commercialization. To address these limitations, DNA Nanobots is developing a novel platform for the safe, affordable, and effective delivery of gene therapies. The Genobots™ platform integrates scaffolded DNA Origami techniques, tissue-targeting antibodies, nuclear- targeting domains, and homology-directed repair mechanisms to achieve safe, efficient, virus-free delivery and integration of a gene payload of any size in a specific target tissue with the option for repeated dosing. In this Phase I SBIR, we propose to demonstrate proof-of-concept for the Genobot platform, with a collection of in vitro and in vivo experiments intended to demonstrate our ability to design Genobots that can accommodate a large range of payload sizes, target specific tissues, and achieve sufficient nuclear penetration and genome integration to be clinically valuable for treating genomic disorders. Aim 1. Construct and characterize Genobots containing payloads of three different sizes. Aim 2. Demonstrate that Genobots bind to, are taken up by, and expressed in target cells but not non-target cells. Aim 3. Demonstrate tissue-targeted gene expression of Genobots in vivo. Go/No-Go Criterion for Progression to Phase II—1) Accumulation of a tissue-targeted Genobot is ≥ 2-fold higher in the target tissue than the accumulation of a non-targeted Genobot, 2) tissue-targeted Genobots do not accumulate in non-target tissues to a greater degree than non-targeted Genobots, and 3) expression of target gene mRNA and expression of the corrected protein are ≥ 2-fold higher in the Genobot-targeted tissue than in non-target tissues. IMPACT – In combination, we expect the key features of our Genobot platform will provide a safe, effective, low-cost virus-free alternative to current gene delivery platforms with the added benefits of improved tissue targeting, larger payload capacity, and the option for safe redosing. We expect that commercialization of this technology would dramatically expand the availability of gene therapies by overcoming the manufacturing challenges of virus-based systems, reducing the cost of manufacturing by up to 90%, and enabling the correction of genes that are larger than 5 kb. Collectively, these advantages have the potential to fulfill the long-sought promise of gene therapy as a safe, effective treatment for thousands of genetic diseases that currently have no disease modifying treatments or cures.