SBIR-STTR Award

Hereditary tyrosinemia type 1 (HT1) is an autosomal recessive Inborn Error of Metabolism (IEM), characterized by the inability to metabolize tyrosine and caused by the deficiency in fumarylacetoacetate hydrolase (FAH) enzyme. The liver is central to IEMs, including HT1. Acute HT1 causes severe liver dysfunction and death, if left untreated. Chronic HT1 leads to liver fibrosis and hepatocellular carcinoma (HCC). Early and life-long pharma- cological treatment with nitisinone (NTBC) can minimize liver damage. However, due to residual cellular toxicity that accumulates over years, HT1 patients on nitisinone remain at risk of cirrhosis, HCC, and liver failure and experience progressive neurocognitive decline. Liver transplants remain the only curative therapy for HT1. Liver-directed gene therapy, in which the defective gene is replaced with a functional isoform, has the potential to replace risky liver transplants and costly life-long pharmacological treatment. HydroGene has developed a non-viral gene delivery to the liver for affordable and safe gene therapy that can meet all the requirements for successful HT1 therapy. Unlike the Lentiviral method, which suffers from low efficacy of liver delivery, safety issues related to systemic immunotoxicity, and expensive manufacturing that limits its scalability, HydroGene has developed a routine method of efficient hydrodynamic delivery of non-viral DNA specifically to hepatocytes via the biliary system. Because the method of gene delivery is based on a routine endoscopic clinical procedure that can be performed in under 30 min and has minimal complications, it is ideal for translation to clinics. In addition, the method can deliver non-immunogenic naked DNA that is cheap to manufacture and can be easily tailored to different rare diseases. Therefore, the HydroGene liver gene therapy is suitable to be translated to clinics for cost-effective life-long correction of HT1 and other rare monogenetic liver diseases. In this Fast-Track application, the objective is to demonstrate the clinically relevant efficacy of using hydrody- namic gene delivery for the treatment of HT1. One of the key features of HT1, and many other rare genetic liver disorders, is continuous liver damage and injury, causing excessive cell death. As a result, corrected hepato- cytes have a survival advantage and will repopulate the damaged liver, but only if the corrective gene is trans- ferred into both daughter cells. Thus, to cure HT1, it is essential for the FAH gene to be integrated into the host genome. To this end, the focus of Phase 1 is to optimize the vector system for safe genome integration and efficient cure in a HT1 mouse model. All the data obtained in Phase 1 is directly translatable to studies in a large animal proposed in Phase 2. The focus of Phase 2 is to determine the exact vector composition and regimen for interruption/tapering treatment with nitisinone in the most clinically relevant large animal HT1 model. The final product of this proposal is a clear path toward clinical translation of hydrodynamic delivery of an integrating vector system to treat HT1 patients and beyond, to many other inborn errors of metabolism and liver diseases.

Public Health Relevance Statement:
NARRATIVE There is currently no gene therapy delivery method that is suitable for translation into the clinic to safely treat Hereditary Tyrosinemia type 1 (HT1). HydroGene has developed a method of hydrodynamic injection through the biliary system to deliver non-viral DNA directly and efficiently into human livers. Given the routine nature and safety of the procedure, along with low costs associated with the production of naked DNA, this method is well suited for clinical translation toward scalable and affordable gene therapy of HT1 and many other genetic liver diseases. Terms: <4, 6-dioxo-heptanoic acid; 4, 6-dioxoheptanoic acid; Acute; Affect; Ammonia; Animal Model; Animal Models and Related Studies; Animals; Benign; Bile Tract; Biliary; Biliary System; Biliary Tree; Biochemical; Biodistribution; Blood; Blood Circulation; Blood Reticuloendothelial System; Bloodstream; Cell Body; Cell Death; Cells; Cessation of life; Chronic; Cirrhosis; Clinic; Clinical; Clinical Trials; DNA; DNA Therapy; DNA Transposons; Data; Death; Deoxyribonucleic Acid; Disease; Disorder; Dose; Enzyme Gene; Enzymes; Family suidae; Fibrosis; Fumarylacetoacetase; Gene Delivery; Gene Expression; Gene Transfer Clinical; Generalized Growth; Genes; Genetic; Genetic Intervention; Genome; Genomics; Goals; Growth; Hepatic Cells; Hepatic Disorder; Hepatic Failure; Hepatic Parenchymal Cell; Hepatic Transplantation; Hepatocarcinoma; Hepatocellular Carcinoma; Hepatocellular cancer; Hepatocyte; Hepatoma; Hereditary; Hereditary Metabolic Disorder; Hereditary Tyrosinemias; Histology; Human; Hydrolase; Hydrolase Family Gene; Hydrolase Gene; IQ Deficit; Immune; Immunes; In Vitro; Inborn Errors of Metabolism; Inflammation; Inherited; Injections; Injury to Liver; Interruption; Isoforms; Left; Legal patent; Life; Liquid substance; Live Birth; Liver; Liver Cells; Liver Cells Carcinoma; Liver Dysfunction; Liver Failure; Liver Fibrosis; Liver Grafting; Liver Transplant; Liver diseases; Membrane; Messenger RNA; Methods; Mice; Mice Mammals; Modeling; Modern Man; Murine; Mus; Nature; Neurocognitive Deficit; Neurologic; Neurological; Non-Polyadenylated RNA; Normal Range; Normal Values; Orphan Disease; Other Genetics; Patents; Patients; Pharmacological Treatment; Phase; Phenotype; Pigs; Population; Primary carcinoma of the liver cells; Procedures; Production; Protein Isoforms; Protocol; Protocols documentation; RNA; RNA Gene Products; Rare Diseases; Rare Disorder; Regimen; Residual; Residual state; Ribonucleic Acid; Risk; Safety; Suidae; Swine; System; Tail; Tissue Growth; Toxic effect; Toxicities; Transgenes; Translating; Translations; Transposase; Treatment Efficacy; Treatment Protocols; Treatment Regimen; Treatment Schedule; Tyrosine; Tyrosine Metabolism; Tyrosine Metabolism Pathway; Tyrosinemias; Veins; Weight; autosome; biliary tract; cirrhotic; clinical applicability; clinical application; clinical relevance; clinical translation; clinically relevant; clinically translatable; cost; cost effective; curative intervention; curative therapeutic; curative therapy; curative treatments; damage to kidney; daughter cell; deliver mRNA; deliver messenger RNA; delivery system for mRNA; experience; fibrotic liver; fluid; fumarylacetoacetate fumarylhydrolase; fumarylacetoacetate hydrolase; gene repair therapy; gene therapy; gene-based therapy; gene-based treatment; gene-directed therapy; gene-targeted therapy; gene-targeted treatment; genetic therapy; genomic therapy; hepatic body system; hepatic damage; hepatic disease; hepatic fibrosis; hepatic injury; hepatic organ system; hepatocyte injury; hepatopathy; immunotoxicity; in vivo; inborn metabolism disorder; intelligence quotient deficit; intervention efficacy; kidney damage; liquid; liver carcinoma; liver damage; liver disorder; liver injury; liver transplantation; mRNA; mRNA delivery; manufacture; meeting; meetings; membrane structure; messenger RNA delivery; model of animal; mouse model; murine model; necrocytosis; neurocognitive decline; neurocognitive impairment; non-viral gene delivery; non-viral gene therapy; nonviral gene delivery; nonviral gene therapy; ontogeny; orphan disorder; plasmid DNA; porcine; pressure; procedure safety; promoter; promotor; renal damage; safe procedure; succinylacetone; suid; therapeutic efficacy; therapy efficacy; toxic reaction in immunology; transgene; translation; transposon element; tumor; vector; weights