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Principal Investigator: Keith Thomas Gagnon
Organization: WAKE FOREST UNIVERSITY HEALTH SCIENCES
Fiscal Year: 2024
Award: $180,537
Funding agency: National Institute of Allergy and Infectious Diseases
PROJECT SUMMARY
Hepatitis B virus (HBV) infection remains an incurable disease. Despite the availability of vaccines and
potent anti-viral drugs, nearly 300 million people worldwide are infected with HBV. The challenge with
curing HBV is the persistence of the virus’ genome in the form of a double-stranded covalently closed
circular DNA, or cccDNA, in the liver cells of patients. Unfortunately, the most modern nucleoside analogs
(NUC) drugs do not eliminate cccDNA and infection is never fully cleared. To ultimately achieve a cure for
HBV, a new class of drugs are needed that can specifically induce degradation or loss of the cccDNA.
This project will address the need for cccDNA targeting drugs by proposing to develop peptide nucleic
acids, or PNAs, as a therapeutic. PNAs are similar to small nucleic acid therapeutics that have already
been FDA approved, like Patisiran or Nusinersen. However, unlike other small nucleic acid drugs, PNAs
have the unique ability to directly bind to double-stranded DNA (dsDNA) and induce damage or
degradation. And unlike some biologics are being developed that can also bind and degrade dsDNA, like
CRISPR-Cas systems or TALENs, PNAs are extremely stable, exhibit low toxicity, are highly sequence
specific, and are deliverable without formulation of as nanoparticle formulations. PNAs can also build on
the successful development of FDA-aproved liver-targeted nucleic acid therapeutics like Patisiran.
This project will first establish sensitive cccDNA detection and characterization methods to track
changes in cccDNA during the course of experimental treatments with PNAs. It will design, synthesize
and test a small library of PNAs that can target and potentially induce the degradation of cccDNA. These
will include two PNA binding modes to the cccDNA and DNA-modifying chemical conjugates, including
DNA alkylation and phosphodiester bond cleavage. These PNAs will be characterized in vitro and in cell
culture models of HBV. The best PNA designs will then be tested in mouse models using two modes of
delivery, direct injection or lipid nanoparticle (LNP) formulation, and their acute toxicity, immunogenicity,
and tissue distribution characterized. Finally, the best PNAs will further characterized for their dosing,
duration of effect, and ability to eliminate cccDNA in two mouse models of HBV. Together, these results
will evaluate the potential of PNAs to serve as a drug candidate for clearing cccDNA from infected animal
models of HBV. If successful, one or more lead PNAs could be identified for immediate preclinical
development to cure HBV, either alone or combined with currently prescribed NUCs or other antivirals.
Terms: <Acceleration><Affect><Animal Testing><Animals><Anti-viral Agents><Binding><Biological Agent><Biological Products><CRISPR><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas system><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cas nuclease technology><Cell Body><Cell Culture Techniques><Cell model><Cells><Cellular model><Chemicals><Chronic><Chronic Hepatitis B><Circular DNA><Cirrhosis><Clampings><Clinical Evaluation><Clinical Testing><Closure by clamp><Clustered Regularly Interspaced Short Palindromic Repeats><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Contracting Opportunities><Contracts><DNA><DNA Alkylation><DNA Binding><DNA Binding Interaction><DNA Damage><DNA Injury><DNA bound><DNA purification><Deoxyribonucleic Acid><Detection><Development><Disease><Disorder><Dose><Double-Stranded DNA><Drug Prescribing><Drug Prescriptions><Drug Targeting><Drug or chemical Tissue Distribution><Drugs><Effectiveness><Exhibits><FDA approved><Formulation><Galactosamine><Gene Transcription><Genetic Transcription><Genome><HBV><HBV Animal Model><HBV infection><Health><Hepatic Cells><Hepatic Cirrhosis><Hepatic Disorder><Hepatic Parenchymal Cell><Hepatitis B><Hepatitis B Infection><Hepatitis B Virus><Hepatocarcinoma><Hepatocellular Carcinoma><Hepatocellular cancer><Hepatocyte><Hepatoma><Homologous Serum Hepatitis Virus><In Vitro><Infection><Injections><Injury to Liver><Invaded><Lead><Libraries><Liver><Liver Cells><Liver Cells Carcinoma><Liver Cirrhosis><Liver diseases><Medication><Methodology><Methods><Mice><Mice Mammals><Modeling><Modernization><Molecular Interaction><Monitor><Murine><Mus><Nucleic Acid Biochemistry><Nucleic Acids><Oligo><Oligonucleotides><Patients><Pb element><Peptide Nucleic Acids><Persons><Pharmaceutical Preparations><Phase><Polymerase><Primary carcinoma of the liver cells><Property><Protocol><Protocols documentation><QOL><Quality of life><RNA Expression><Recommendation><Solid><System><TAL effector nuclease><TAL endonuclease><TALE nuclease><TALEN technology><TALENs><Tail><Testing><Therapeutic><Therapeutic Effect><Tissue Distribution><Toxic effect><Toxicities><Transcription><Vaccines><Viral><Viral Antigens><Viral Diseases><Viral Genome><Viral Hepatitis B><Virus Diseases><Wild Type Mouse><access to vaccination><access to vaccines><acute toxicity><anti-viral compound><anti-viral drugs><anti-viral medication><anti-viral therapeutic><anti-virals><biologics><biopharmaceutical><biotherapeutic agent><cell culture><cell cultures><chemical conjugate><chemical synthesis><chronic HBV infection><chronic hepatitis B virus infection><cirrhotic><clinical test><curative intervention><curative therapeutic><curative therapy><curative treatments><design><designing><determine efficacy><developmental><drug candidate><drug/agent><ds-DNA><dsDNA><efficacy analysis><efficacy assessment><efficacy determination><efficacy evaluation><efficacy examination><evaluate efficacy><examine efficacy><functional group><heavy metal Pb><heavy metal lead><hepatic body system><hepatic damage><hepatic disease><hepatic injury><hepatic organ system><hepatitis B virus animal model><hepatopathy><immunogenicity><in vitro testing><infected with HBV><infected with hepatitis B><infected with hepatitis B virus><infection with HBV><infection with hepatitis B virus><lead candidate><lipid based nanoparticle><lipid nanoparticle><liver carcinoma><liver damage><liver disorder><liver infection><liver injury><medication prescription><mouse model><murine model><nano particle><nano pore><nano-sized particle><nanoparticle><nanopore><nanosized particle><new drug class><novel><novel drug class><nucleic acid analog><nucleic acid therapy><nucleic acid-based therapeutics><nucleoside analog><oligos><phosphodiester><pre-clinical development><preclinical development><prescribed medication><prevent><preventing><research clinical testing><serum hepatitis><site targeted delivery><success><targeted delivery><therapeutic nucleic acids><transcription activator-like effector nucleases><vaccination access><vaccination availability><vaccine access><vaccine availability><viral genomics><viral infection><virology><virus antigen><virus genome><virus genomics><virus infection><virus-induced disease><wildtype mouse>