To commercialize non-viral gene medicines, it is critical that both vector potency (i.e. therapeutic transgene expression levels) and the duration of the therapeutic effect be improved. Potent dose-sparing extended duration gene therapies will have a cost and efficacy competitive advantage over alternative technologies. In this Phase I proof of concept study, we will create a novel antibiotic-free MiniPlasmid gene therapy platform for extended duration gene therapy. The vectors combine transient expression enhancers that improve transgene expression levels with a novel 270 base pair replication origin-antibiotic free selection cassette that we hypothesize will promote long duration gene expression after vector delivery to the body. The MiniPlasmid platform will be applied to create a wound healing gene therapy product to treat diabetic neuropathic foot ulcers. In Specific Aims 1 and 2 a high yielding MiniPlasmid fermentation manufacturing platform is created. In Specific Aim 3 the MiniPlasmid vector platform is validated in vivo for extended duration expression compared to conventional plasmids. A hypoxia- inducible factor 1¿ (HIF-1 ¿) based gene medicine for diabetic foot ulcer treatment is developed utilizing an extended half-life oxygen resistant highly active HIF-1 ¿ mutant (CA5-HIF-1 ¿). Specific Aim 3 is performed in collaboration with wound healing gene therapy expert Dr. John Harmon at Johns Hopkins University. The MiniPlasmid vector platform is designed to improve transgene expression levels and duration to enable gene medicine development for multiple applications requiring extended duration expression. MiniPlasmid vectors developed in Phase I will be marketed to investigators for a variety of gene therapy applications through publications, trade shows, and the Nature Technology Corporation (NTC) website. In Phase II the HIF-1 ¿ MiniPlasmid gene therapeutic will undergo preclinical safety and efficacy evaluations for treatment of diabetic foot ulcers prior to clinical development in Phase III.
Public Health Relevance Statement: Public Health Relevance: The objective of this proposal is to develop a novel antibiotic-free non-viral MiniPlasmid gene therapy platform for extended duration gene therapy, and as such is responsive to NIGMS SBIR high-priority area of interest in development of improved vectors for gene transfer. The vectors combine transient expression enhancers that improve transgene expression levels with a novel replication origin-antibiotic free selection cassette that affords long duration gene expression after gene delivery to the body. The platform will be applied to create a wound healing gene therapy product to treat diabetic neuropathic foot ulcers.
NIH Spending Category: Biotechnology; Diabetes; Gene Therapy; Genetics; Neurodegenerative; Neuropathy
Project Terms: American; Antibiotics; Area; base; Base Pairing; Biodistribution; Biology; Blood Coagulation Factor; Cell Line; Clinical; Collaborations; Combined Antibiotics; commercialization; cost; design; Development; Diabetes Mellitus; diabetic; Diabetic Foot Ulcer; Digestion; Dose; Drug Kinetics; efficacy evaluation; Electroporation; Enhancers; Evaluation; Excision; expression vector; Factor IX; Factor VIII; Fermentation; Foot Ulcer; Gene Delivery; Gene Expression; Gene Silencing; gene therapy; Gene Transduction Agent; gene transfer vector; Genes; Genetic Recombination; Half-Life; Hemophilia A; Hemophilia B; Heterochromatin; Human; Hypoxia Inducible Factor; hypoxia inducible factor 1; improved; In Vitro; in vivo; innovation; interest; Interleukin-10; Intramuscular; Letters; Libraries; limb amputation; Lower Extremity; Malignant Neoplasms; Marketing; Mediating; Medicine; Methylation; Mus; Mutagenesis; mutant; Mutation; National Institute of General Medical Sciences; Nature; Neuropathy; non-viral gene therapy; novel; Oxygen; painful neuropathy; Phase; Plasmid Cloning Vector; Plasmids; preclinical efficacy; preclinical safety; Prevention; Process; Production; Promotor (Genetics); Proteins; public health medicine (field); public health relevance; Publications; Replacement Therapy; Replication Origin; Replicon; Reporter; Research Personnel; Resistance; RNA; RNA Interference; Site; Small Business Innovation Research Grant; Solutions; Technology; Testing; Therapeutic Effect; therapeutic gene; therapeutic transgene; therapy development; Time; Transfection; transgene expression; Universities; vector; Vertebral column; web site; Wound Heali
