The goal of this study is to develop an RNAi-based genetic precision medicine to treat alcohol associated liver disease (AALD). AALD is a devastating health problem worldwide, accounts for the majority of alcohol-related mortality globally and is the second most indication for liver transplantation in the US. The current medical management for AALD remains limited, and no proven pathobiology-driven pharmacotherapy is available. There is thus an urgent need to identify novel targets and develop promising therapies for both prevention and treatment for AALD. Recent genome-wide association studies (GWAS) has identified that a single nucleotide polymorphism (SNP rs738409 C>G, or amino acid alteration at position 148 I>M) in the patatin-like phospholipase domain containing 3 (PNPLA3) gene is the single largest genetic risk factor contributing to the progression of AALD to advanced liver perturbations including liver cancer. The PNPLA3 148 I>M polymorphism is the molecular reason why this subset of AALD patients develop severe liver injuries. Mechanistic studies have revealed that overexpression of the mutant PNPLA3 (the 148M isoform) is underlying both liver steatosis and fibrosis and inflammation by interrupting hepatic triglyceride metabolism. Our creation of a humanized PNPLA3148M mouse model successfully recapitulates the AALD phenotypes and has corroborated the aforementioned mechanism. We hypothesize that transcriptional downregulation of the PNPLA3148M isoform is a cause-targeting strategy for the treatment of AALD with the PNPLA3148M allele. To this end, we pioneered the development and have issued patents for a precision allele-specific small interfering RNA (siRNA) that possesses excellent specificity for the PNPLA3148M isoform compared to its wild-type counterpart. We further discovered two more modified oligonucleotides which significantly improved the medicinal chemistry specifications of our therapeutic lead. Administration of our lead siRNA to the liver of the humanized PNPLA3148M model fed an ethanol-containing diet significantly reduced the PNPLA3148M expression as well as hepatic steatosis, inflammation and fibrosis. We also demonstrated that delivering our siRNA into the humanized PNPLA3148M model with the N-Acetylgalactosamine (GalNAc)-conjugation, a well-established liver-specific drug delivery strategy, effectively knocked down hepatic hPNPLA3148M mRNA and ameliorated the liver histology. With this success, we propose to advance our drug candidate development by focusing on an Investigational New Drug (IND) application to the FDA. Specifically, we aim: 1) to define the best drug candidate of our hPNPLA3148M-targeting, GalNAc-conjugated oligonucleotides (Phase I); 2) to collect IND-enabling quality assurance and safety data in rodents and non-human primates (Phase II). We expect that via this study, we will develop the first genetic medicine for AALD treatment, which could also serve as a new prototype for drug development for other common human diseases. The collected data in this study will enable us to file the IND application to support our first clinical trial in humans.
Public Health Relevance Statement: PROJECT NARRATIVE Alcohol associated liver disease (AALD) is one of the major liver diseases with a high mortality rate but no effective drug treatment for AALD at present. Our project for the first time aims at developing a personalized genetic medicine for AALD, which has a great impact to both improving the human health and reducing social burden due to this deadly disease. We use novel and unprecedented approach in this project, which has a great potential to establish a new paradigm for drug develop for other common diseases.
Project Terms: N acetylgalactosamine; Acetylgalactosamine; Alcohol Drinking; EtOH drinking; EtOH use; alcohol ingestion; alcohol intake; alcohol product use; alcohol use; alcoholic beverage consumption; alcoholic drink intake; ethanol consumption; ethanol drinking; ethanol ingestion; ethanol intake; ethanol product use; ethanol use; Alcohol consumption; Absolute ethanol; ETOH; Ethyl Alcohol; Grain Alcohol; Methylcarbinol; Ethanol; Allelomorphs; Alleles; aminoacid; Amino Acids; Animals; Chemistry; Pharmaceutical Chemistry; Medicinal Chemistry; Pharmaceutic Chemistry; Clinical Research; Clinical Study; Clinical Trials; Cessation of life; Death; Diet; diets; Disease; Disorder; DNA; Deoxyribonucleic Acid; Down-Regulation; Pharmacotherapy; Drug Therapy; drug treatment; Pharmaceutical Preparations; Drugs; Medication; Pharmaceutic Preparations; drug/agent; Investigational Drugs; Investigational New Drugs; Fatty Liver; Liver Steatosis; hepatic steatosis; hepatosteatosis; Fibrosis; Future; Genes; Goals; Health; Histology; Human; Modern Man; Inflammation; Lead; Pb element; heavy metal Pb; heavy metal lead; Liver; hepatic body system; hepatic organ system; Liver diseases; Hepatic Disorder; hepatic disease; hepatopathy; liver disorder; Alcoholic Liver Diseases; alcohol induced hepatic injury; alcohol induced liver disorder; alcohol induced liver injury; alcohol related liver disease; alcohol-associated liver disease; alcohol-induced hepatic dysfunction; alcohol-induced liver disease; alcohol-induced liver dysfunction; alcohol-mediated liver dysfunction; alcohol-mediated liver injury; alcohol-related liver disease; alcoholic liver injury; ethanol induced hepatic injury; ethanol induced liver disorder; ethanol induced liver injury; ethanol liver disease; ethanol-induced hepatic dysfunction; ethanol-induced liver disease; ethanol-induced liver dysfunction; ethanol-mediated liver dysfunction; ethanol-mediated liver injury; liver transplantation; Hepatic Transplantation; Liver Grafting; Liver Transplant; Metabolism; Intermediary Metabolism; Metabolic Processes; Monkeys; mortality; Mutation; Genetic Alteration; Genetic Change; Genetic defect; genome mutation; Names; name; named; naming; Oligonucleotides; Oligo; oligos; Legal patent; Patents; Patients; Drug Kinetics; Pharmacokinetics; Pharmacology; Phenotype; Phospholipase; Lecithinases; Plasma; Blood Plasma; Plasma Serum; Reticuloendothelial System, Serum, Plasma; Genetic Polymorphism; polymorphism; Proteins; Rattus; Common Rat Strains; Rat; Rats Mammals; Messenger RNA; mRNA; Rodent; Rodentia; Rodents Mammals; Role; social role; Safety; Specificity; Testing; Time; Drug or chemical Tissue Distribution; Tissue Distribution; Toxicology; Genetic Transcription; Gene Transcription; RNA Expression; Transcription; Antisense Oligonucleotides; Anti-Sense Oligonucleotides; Antisense Agent; anti-sense agent; anti-sense oligo; antisense oligo; Drug Delivery; Drug Delivery Systems; Investigational New Drug Application; quality assurance; improved; Hepatic; Chronic; Clinical; Specified; Specific qualifier value; Phase; Variation; Variant; Medical; Evaluation; Susceptibility; Predisposition; Hepatotoxic effect; Liver Toxicity; Toxic effect on liver cells; hepatic toxicity; hepatoxicity; Hepatotoxicity; Individual; non-human primate; nonhuman primate; fibrotic liver; hepatic fibrosis; Liver Fibrosis; Therapeutic; Genetic; Malignant neoplasm of liver; Hepatic Cancer; liver cancer; liver malignancy; malignant liver tumor; programs; Investigation; meetings; meeting; mutant; Protein Isoforms; Isoforms; success; genotoxicity; Toxic effect; Toxicities; novel; AODR mortality; AODR death rate; Alcohol or Other Drug Related death rate; Alcohol or Other Drug Related mortality; alcohol related mortality; Prevention; social; Positioning Attribute; Position; Single Nucleotide Polymorphism; Single Base Polymorphism; single nucleotide variant; genetic risk factor; Genetic predisposing factor; inherited factor; Modeling; drug development; disorder subtype; disease subgroups; disease subtype; alcohol abuse therapy; alcohol abuse treatment; alcohol treatment; Manufacturer; Short interfering RNA; siRNA; Small Interfering RNA; Post-Transcriptional Gene Silencing; Posttranscriptional Gene Silencing; Quelling; RNA Silencing; RNAi; Sequence-Specific Posttranscriptional Gene Silencing; RNA Interference; Triglyceride Metabolism; Causality; causation; disease causation; Etiology; Data; Dominant-Negative Mutation; Antimorphic mutation; Dominant Negative; Dominant-Negative Mutant; Dose Limiting; Interruption; Research Contracts; Genetic Medicine; Molecular; Modification; Development; developmental; disease phenotype; GWA study; GWAS; genome wide association; genome wide association scan; genome wide association studies; genomewide association scan; genomewide association studies; genomewide association study; whole genome association analysis; whole genome association studies; whole genome association study; genome wide association study; knockdown; knock-down; immunogenicity; designing; design; human study; shRNA; short hairpin RNA; small hairpin RNA; human disease; new drug treatments; new drugs; new pharmacological therapeutic; new therapeutics; new therapy; next generation therapeutics; novel drug treatments; novel drugs; novel pharmaco-therapeutic; novel pharmacological therapeutic; novel therapy; novel therapeutics; murine model; mouse model; therapeutic target; prototype; NASH; non-alcohol induced steatohepatitis; non-alcoholic steato-hepatitis; non-alcoholic steatohepatitis; nonalcoholic steato-hepatitis; nonalcoholic steatohepatitis; overexpress; overexpression; FDA approved; treatment strategy; safety testing; good laboratory practice; comparable efficacy; compare efficacy; comparative efficacy; drug candidate; Phase I Study; phase 1 study; phase II study; phase 2 study; screenings; screening; Injury to Liver; hepatic damage; hepatic injury; liver damage; liver injury; precision-based medicine; precision medicine; personalized genetics; precision genetics; precision therapies; precision treatment; Precision therapeutics; targeted drug therapy; targeted drug treatments; targeted therapeutic; targeted therapeutic agents; targeted therapy; targeted treatment; Formulation; therapeutic toxicity; therapy associated toxicity; therapy related toxicity; therapy toxicity; treatment toxicity; treatment-associated toxicity; Treatment-related toxicity; first in man; first-in-human; Good Manufacturing Process; Good manufacturing practice; manufacture
