Advancing manufacturing and release testing of a gene editing therapy for Duchenne muscular dystrophy

Project Summary Optimization of the manufacturing process and analytical testing for adeno-associated virus (AAV) products is important to put in place as early as possible. Being able to use the same process and release testing throughout development will allow bridging of nonclinical and clinical study material and pooling of data from clinical trials. It is critical to begin work early on analytical development to have assays in place prior to manufacture of cGMP material and IND filing. Here, a dual AAV vector gene editing therapy is being developed for Duchenne muscular dystrophy. In order to include Cas9 inactivation and encompass the large size of Cas9, two vectors are used with one carrying Cas9 and one carrying the guide RNAs. In this proposal, a Design of Experiment (DoE) optimization will be performed on the upstream manufacturing process for both vectors with the aim of improving yield and quality of material. Then product-specific analytics will be optimized and other standard non-compendial assays tested on the two vectors. We will utilize technical assistance from an experienced AAV contract development and manufacturing organization and CMC and regulatory consultants to ensure the work is conducted to regulatory standards. This project will finalize standard operating procedures for manufacturing and release testing of the gene editing therapy for Duchenne. Thus, this work is critical to support the advancement of this dual vector gene editing therapy to the clinic and then commercialization. This therapy is impactful because it targets deletion of DMD exons 45-55, thereby restoring the reading frame for 50% of Duchenne muscular dystrophy patients who have no cure and only limited approved therapies.