Platform for Optimizing Yield and Quality of Therapeutic AAV Vector Manufacturing via Synthetic Helper Genes

PROJECT SUMMARY In this Direct-to-Phase II SBIR, Synvivia, Inc. proposes to validate a platform for identifying cyclic peptides (CPs) that dramatically increase the yield and quality of adeno-associated virus (AAV) vectors manufactured for gene therapy. Problem: AAV-based gene therapies hold great promise for preventing, treating, or curing a wide range of inherited disorders and chronic illnesses, but the complexity and cost of manufacturing is a major barrier to realizing the full promise of this technology. Current manufacturing is constrained by 1) poor scalability of cell culture systems, 2) low yield of functional vectors, 3) high loss of functional vectors in downstream processing, and 4) limited mechanisms for optimizing product yield or quality. These constraints drive up the cost of goods sold, limit production to levels that can only serve rare indications, and increase the risk of adverse effects. Solution: Synvivia has developed a massively multiplexed synthetic helper gene cell-control strategy that is designed to identify and deliver molecules to vector packaging cells that optimize the cell’s state for biomanufacturing. The platform uses a competitive enrichment process to identify members of a cyclic peptide (CP) library that optimize vector yield and/or quality in a manufacturer’s specific vector manufacturing process, overcoming the industry-wide challenge posed by disparate methods and materials. Preliminary Data: Synvivia developed a ~50M-member CP Discovery Library and demonstrated proof-of-concept for identifying members of the library that increase AAV physical titer, functional titer, and transduction efficiency in the context of three clinically-relevant AAV serotypes. Out of thousands of CPs identified with potentially beneficial effects on vector yield and quality, 36 were randomly selected for further characterization. Most of the selected CPs increased both physical titer and functional titer by up to 10-fold, and most eliminated 90% to 95% of all full but non- functional capsids, which improved AAV transduction efficiency by up to 17-fold. These gains should translate to more doses per batch, a lower cost of goods sold, and improved patient safety. Proposed Work: Synvivia proposes to validate the platform by demonstrating three things manufacturers have requested: 1) CP Discovery Library contains members relevant to a broader array of AAV serotypes, 2) CPs discovered at bench-scale improve vector yield and quality at bioreactor scale, and 3) vectors produced through CP-enhanced manufacturing perform better than vectors from a standard process when administered in vivo. Aim 1. Demonstrate Synvivia’s CP Discovery Library contains CPs that improve the yield and quality of AAV vector production across multiple manufacturing contexts. Aim 2. Validate lead CPs in vitro and in vivo. Impact: This study will provide the data that manufacturers need before committing to Synvivia’s CP discovery process. CP- enhanced production is expected to substantially improve vector yield and quality, thereby increasing production, reducing costs, and improving the affordability and safety of AAV-based therapies.