?The development of transformative therapeutics, including the possibility of a cure though gene therapy, continues to be the major research and development activity in the commercial hemophilia space. For example, there are several company-sponsored ongoing clinical trials of recombinant adeno-associated viral (rAAV) encoding coagulation factor IX (FIX) for the treatment of hemophilia A. Furthermore as a result of these trials, there now exists extensive safety and initial efficacy data for multiple rAAV-FIX products. However, progress in the translation of this approach to hemophilia A has been hampered by significant hurdles including the size, complexity, immunogenicity, instability and biosynthetic inefficiency of coagulation factor VIII (FVIII) in comparison to FIX. Now through the identification of 1) liver-specific transcriptional regulatory modules (LTRMs) that are smaller tha existing enhancer-promoters combinations and 2) a liver-enhanced codon- optimization algorithm used to engineer the coagulation factor VIII (FVIII) transgene, we have been able to design rAAV vectors encoding FVIII with genome sizes under 5,000 nucleotides that have unprecedented potency. The proposed milestones of the current project are 1) to characterize and select a lead candidate LTRM optimized for FVIII mRNA and transgene product output, rAAV vector manufacture yield and rAAV product consistency, and 2) to identify a lead candidate AAV-FVIII transgene. These proof of concept studies will be performed using the preclinical murine model of hemophilia A. The data obtained should facilitate the selection and preclinical validation of a lead candidate liver-directed AAV-FVIII expression cassette combining these novel technologies exclusively available to Expression Therapeutics (ET). Therefore, ET is in an opportunistic position to capitalize on the ongoing rAAV-FIX/hemophilia B clinical findings and incorporate preclinical R&D and clinical protocol development strategies into this high priority rAAV-FVIII project. This existing clinical gene therapy knowledge combined with our hemophilia A experience and innovative technologies should enable ET to overcome the major hurdles facing commercialization of liver-directed rAAV-FVIII therapy for hemophilia A.
Public Health Relevance Statement: Public Health Relevance: The goal of this project is test new technologies developed initially for liver-directed recombinant adeno-associated viral vector gene therapy of hemophilia A. Our group has developed both liver expression and size optimized transcription regulatory modules and a liver-optimized coagulation factor VIII transgene that should overcome the existing barriers to rAAV-based gene therapy of hemophilia A. The preclinical data obtained from the proposed studies should facilitate the selection of a lead gene therapy vector candidate.
Project Terms: Address; adeno-associated viral vector; Algorithms; Amino Acid Sequence; Amino Acids; Anabolism; B-domain-deleted factor VIII; Biomedical Engineering; Capsid; Clinical; Clinical Data; Clinical Protocols; Clinical Trials; Codon Nucleotides; combinatorial; commercialization; Complementary DNA; Computer Simulation; cytotoxic; Data; design; Development; Disease; DNA cassette; Dose; Engineering; Enhancers; experience; Factor IX; Factor VIII; Foundations; gene therapy; Gene Transduction Agent; Gene Transfer; Genetic Transcription; Genome; Goals; Hemophilia A; Hemophilia B; Human; Immune; Immunity; immunogenicity; improved; In Vitro; in vivo; Injection of therapeutic agent; innovative technologies; Intellectual Property; Inverted Terminal Repeat; Knowledge; Lead; Liver; Mediating; Messenger RNA; Methods; mouse model; Mus; new technology; novel; Nucleotides; Output; particle; Patients; Peripheral; Phase; plasmid DNA; Polyadenylation; Positioning Attribute; pre-clinical; pre-clinical research; product development; Program Development; promoter; Property; protein expression; protocol development; public health relevance; Publications; Recombinant Proteins; Recombinants; Regulatory Element; Reporting; research and development; Risk; Safety; Small Business Innovation Research Grant; success; System; Technology; Testing; Therapeutic; Time; Transgenes; Translations; Validation; vector; Veins; Viral; Viral Genes; Viral Genome; Viral Vector
