Despite recent advances, development of nucleic acid (NA) therapies continues to be impeded by the challenges of effective delivery. GeneSegues, Inc. (GSI) has developed a nonviral delivery technology, sub-50 nanometer (s50) capsules, which co-opt the raft/caveolar uptake path for efficient intracellular delivery of NA drugs. The raft/caveolar path is an endogenous, intracellular transport mechanism that can efficiently deliver internalized complexes into the cell, avoiding the inflammation and drug degradation mechanisms that have consistently thwarted effective delivery of NA drugs. Exploiting this receptor-mediated, size-sensitive path is achieved with a novel capsule design comprised of a single-layer crystallized polymer shell which shields the NA drug, sterically stabilizes the shell, and targets the capsule. In liver, GSI has demonstrated initial proof-of-concept for liver-targeted, cell-subtype delivery of functional plasmid DNA. Hepatocyte-specific delivery in mice was demonstrated with s50 capsules employing asialoorosomucoid (ASOR) ligands targeting the asialoglycoprotein receptor (ASGPr), while delivery to LSEC's was achieved using hyaluronan (HA)-coated s50 capsules targeting the HA receptor for endocytosis (HARE). We now propose to initiate preclinical development of these liver-targeted, plasmid-bearing s50 capsules for therapeutic application. Additionally, in view of the promising therapeutic potential of RNAi, we propose to also synergistically assess the utility of s50 capsules for systemic in vivo delivery of therapeutic RNAi oligonucleotides to defined liver target sites. Therefore, our proposed specific aims are to (1) determine utility and safety of s50 hepatocyte-targeted capsules for systemic delivery of double and single stranded RNAi oligonucleotides, using a published siRNA sequence (siApoB-1) encapsulated in s50 ASOR-ligand capsules and (2) assess safety of LSEC-targeted s50 capsules for systemic delivery of plasmid DNA, using an epitope-tagged reporter gene encapsulated in s50 HA-ligand capsules. Biodistribution will be assessed by radiolabel for NA's delivered via HA-coated as well as ASOR-coated s50 capsules, and any evidence of early inflammatory responses will be assessed for plasmid and RNAi cargos. If successful, we expect this body of work will provide a solid framework for partnering with NIH and/or drug developers to move forward encapsulated NA therapies for liver disease.
Public Health Relevance: Nucleic acid drugs offer much promise for treating diseases of the liver. However, delivering nucleic acids into disease cells of the liver is very problematic because (1) the liver is a heterogeneous organ with many different cell types, and (2) nucleic acids can trigger drug degradation and toxic reactions once inside the cell. We propose to further develop nucleic acid delivery technology capable of accessing and penetrating specific cell types within the liver, and of avoiding intracellular defenses, for effective treatment of liver disease.
Public Health Relevance Statement: Project narrative Nucleic acid drugs offer much promise for treating diseases of the liver. However, delivering nucleic acids into disease cells of the liver is very problematic because (1) the liver is a heterogeneous organ with many different cell types, and (2) nucleic acids can trigger drug degradation and toxic reactions once inside the cell. We propose to further develop nucleic acid delivery technology capable of accessing and penetrating specific cell types within the liver, and of avoiding intracellular defenses, for effective treatment of liver disease.
NIH Spending Category: Biotechnology; Digestive Diseases; Genetics; Liver Disease
Project Terms: ASGP-R; Achievement; Achievement Attainment; Advanced Development; Adverse effects; Antigenic Determinants; Apo-B; ApoB; Apolipoproteins B; Asialoglycoprotein Receptor; Asialoorosomucoid-Binding Protein; B cell differentiation factor; B cell stimulating factor 2; B-Cell Differentiation Factor-2; B-Cell Stimulatory Factor-2; BCDF; BSF-2; BSF2; BSF2 (B cell stimulating factor 2); Binding Determinants; Biodistribution; Body Tissues; CAPS; CD44 Adhesion Receptor; CD44 Antigens; CD44 molecule; Capsules; Carbohydrates; Cells; Chemistry; Clinic; Collaborations; Complex; DNA; Deoxyribonucleic Acid; Development; Differentiation Factor, B-Cell; Disease; Disorder; Dose; Drug Formulations; Drug Therapy; Drugs; Elements; Encapsulated; Endocytosis; Epitopes; Feasibility Studies; Formulation; Formulations, Drug; Gene Expression; Gene Inactivation; Gene Products, RNA; Gene Silencing; Gene Transfer Clinical; Gene Transfer Procedure; Gene-Tx; Genes; Genetic; Genetic Intervention; Glycoproteins; Goals; Graft Rejection; HPGF; HUTCH-1; Hepatic; Hepatic Cells; Hepatic Cirrhosis; Hepatic Disorder; Hepatic Parenchymal Cell; Hepatocyte; Hepatocyte-Stimulating Factor; Hereditary; Hermes Antigen; Hyaluronan; Hyaluronan-Binding Protein; Hyaluronic Acid Binding Protein; Hybridoma Growth Factor; IFN-beta 2; IFNB2; IL-6; IL6 Protein; INFLM; Immune; Inflammation; Inflammatory Response; Inherited; Interleukin 6 (Interferon, Beta 2); Interleukin-6; Intervention, Genetic; Intracellular Transport; Knowledge; Lead; Ligands; Lipid Rafts, Cell Membrane; Liver; Liver Cells; Liver Cirrhosis; Liver diseases; MGI-2; Mammals, Mice; Mediating; Medication; Membrane Microdomains; Metabolic Diseases; Metabolic Disorder; Methods; Mice; Microscopy; Molecular Biology, Gene Therapy; Molecular Target; Murine; Mus; Myeloid Differentiation-Inducing Protein; NIH; National Institutes of Health; National Institutes of Health (U.S.); Neutron Activation Analyses; Neutron Activation Analysis; Nuclear; Nucleic Acids; Oligo; Oligonucleotides; Organ; Pathway interactions; Pb element; Performance; Pharmaceutic Preparations; Pharmaceutical Preparations; Pharmacotherapy; Phase; Plasmacytoma Growth Factor; Plasmids; Polymers; Publishing; RNA; RNA Sequences; RNA, Non-Polyadenylated; RNA, Small Interfering; RT-PCR; RTPCR; Radiolabeled; Reaction; Receptor Protein; Receptor, Asialoorosomucoid; Receptors, Hyaluronan; Recruitment Activity; Reporter Genes; Research; Reverse Transcriptase Polymerase Chain Reaction; Ribonucleic Acid; Safety; Science of Chemistry; Site; Small Interfering RNA; Solid; Specificity; Sphingolipid Microdomains; Sphingolipid-Cholesterol Rafts; Technology; Therapeutic; Therapy, DNA; Thesaurismosis; Tissues; Toxic effect; Toxicities; Transplant Rejection; Transplantation Rejection; Treatment Side Effects; United States National Institutes of Health; Viral Vector; Work; asialo-orosomucoid; asialoorosomucoid; base; body system, hepatic; capsule (pharmacologic); cell type; clinical relevance; clinically relevant; cytokine; design; designing; disease/disorder; drug/agent; effective therapy; gene therapy; genetic therapy; gp85; heavy metal Pb; heavy metal lead; hepatopathy; in vivo; interferon beta 2; lipid raft; liver disorder; meetings; metabolism disorder; nano meter; nanometer; neoplastic; non-viral gene therapy; nonviral gene therapy; novel; organ system, hepatic; pathway; plasmid DNA; pre-clinical; preclinical; public health relevance; radiolabel; radiotracer; receptor; recruit; response; reverse transcriptase PCR; siRNA; side effect; site targeted delivery; success; targeted delivery; therapy adverse effect; transgene expression; treatment adverse effect; treatment strategy; uptake
