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Principal Investigator: KEITH R JEROME
Organization: FRED HUTCHINSON CANCER CENTER
Fiscal Year: 2024
Award: $862,440
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary
Chronic Hepatitis B virus (HBV) infection (CHB) affects hundreds of millions of people across
the world. Despite effective vaccines, and antiviral therapies that can suppress viral replication,
these infections are largely incurable, and new therapies for CHB are desperately needed. We
have investigated gene editing using targeted nucleases to cleave covalently closed circular
DNA (cccDNA), the template for HBV replication, which results in its degradation or inactivation
in infected hepatocytes. Our work and the work of others has been hindered due to the fact that
existing small animal models for HBV have substantial limitations, or are complex and
expensive to establish. To this end, we established and validated a robust and low-cost liver
humanized NSG-PiZ mouse as a model for the study of HBV. Here we propose to use these
mice for the evaluation of novel curative gene editing therapies that target CHB.
Specific Aim 1. Evaluate the anti-HBV efficacy of HBV gene therapy in liver-humanized
NSG-PiZ mice. We will first evaluate the efficiency of HBV gene editing in NSG-PiZ mice, using
AAV-SaCas9 vectors we previously validated in HBV-infected humanized FRG mice. We will
then compare lipid nanoparticle (LNP) delivery of nuclease mRNA with AAV vectors for the
transduction of PHH in our model. We will compare the antiviral efficacy of different classes of
gene editing nuclease in our NSG-PiZ mouse model of CHB, and evaluate chromatin modifying
agents for their ability to make cccDNA accessible to gene editing nucleases in vitro and in vivo.
Finally, we will assess combined gene-editing, reverse transcriptase inhibitor (RTi), and entry
inhibitor antiviral therapy in our model of CHB.
Specific Aim 2. Evaluate the tolerability and host genomic consequences of AAV- and
LNP-delivered nuclease therapies targeting HBV in liver-humanized NSG-PiZ mice. We will first
determine the maximum tolerated dose (MTD) and minimal effective dose for AAV-nuclease
and LNP-nuclease therapies in our CHB model. We will then monitor the effects of different
nuclease classes on treatment-related off-target genotoxicity in vivo and determine whether use
of liver-specific promoters can mitigate therapy-associated genotoxicity. We will also determine
whether therapy-associated genotoxicity is reduced when nuclease expression is transiently
provided by LNPs. Finally, we will evaluate therapy-specific immune responses and therapy-
associated hepatotoxicity and neurotoxicity after transient or persistent nuclease expression in
immunocompetent mice.
Terms: <2019-nCoV vaccine><AAV delivered><AAV delivery><AAV vector><AAV-based delivery><AAV-based vector><AAV-based viral delivery><AAV-mediated delivery><Address><Adeno-associated-virus-based delivery><Affect><Animal Model><Animal Models and Related Studies><Animals><Anti-viral Agents><Anti-viral Therapy><Apes><Asian><COVID-19 vaccine><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><Cells><Cessation of life><Chromatin><Chronic><Chronic Hepatitis B><Circular DNA><Cirrhosis><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><Complex><DNA><DNA Therapy><Data><Death><Deoxyribonucleic Acid><Development><Dose><Drugs><Enzyme Gene><Enzymes><Evaluation><Event><GUIDE-seq><Gene Combinations><Gene Delivery><Gene Transfer Clinical><Genes><Genetic><Genetic Intervention><Genome><Genome-wide Unbiased Identification of DSBs Enabled by sequencing><Genomics><HBV><HBV Animal Model><HBV infection><HSV><Health><Hepatic Cancer><Hepatic Cells><Hepatic Failure><Hepatic Parenchymal Cell><Hepatitis B><Hepatitis B Infection><Hepatitis B Virus><Hepatocarcinoma><Hepatocellular Carcinoma><Hepatocellular cancer><Hepatocyte><Hepatoma><Hepatotoxic effect><Hepatotoxicity><Herpes Simplex Virus><Herpes labialis Virus><Homologous Serum Hepatitis Virus><Human><Immune mediated therapy><Immune response><Immunocompetent><Immunological response><Immunologically Directed Therapy><Immunotherapy><In Vitro><Infection><Liver><Liver Cells><Liver Cells Carcinoma><Liver Failure><Liver Toxicity><Longitudinal Studies><Malignant neoplasm of liver><Maximal Tolerated Dose><Maximally Tolerated Dose><Maximum Tolerated Dose><Mediating><Medication><Messenger RNA><Mice><Mice Mammals><Modeling><Modern Man><Monitor><Mouse Strains><Murine><Mus><Persons><Pharmaceutical Preparations><Pongidae><Primary carcinoma of the liver cells><Publishing><Reporting><Reverse Transcriptase Inhibitors><Reverse Transcription><Risk><SARS-CoV-2 vaccine><SARS-coronavirus-2 vaccine><Safety><Severe Acute Respiratory Syndrome CoV 2 vaccine><Severe acute respiratory syndrome coronavirus 2 vaccine><Simplexvirus><Study models><Symptoms><System><Therapeutic><Therapeutic Gene Editing><Toxic effect><Toxic effect on liver cells><Toxicities><Transfection><Treatment Efficacy><Tree Shrews><Treeshrews><Tupaiidae><Vaccines><Viral><Viral Diseases><Viral Genes><Viral Hepatitis B><Viremia><Virus><Virus Diseases><Virus Replication><Work><adeno-associated viral vector><adeno-associated viral vector delivery><adeno-associated virus delivery><adeno-associated virus mediated delivery><adeno-associated virus vector><adenovirus mediated delivery><anti-hepatitis B><anti-viral compound><anti-viral drugs><anti-viral efficacy><anti-viral medication><anti-viral therapeutic><anti-virals><chronic HBV infection><chronic hepatitis B virus infection><cirrhotic><clinical relevance><clinically relevant><comparative><coronavirus disease 2019 vaccine><coronavirus disease-19 vaccine><cost><curative intervention><curative therapeutic><curative therapy><curative treatments><delivered with AAV><delivery vector><delivery vehicle><delivery with AAV><determine efficacy><developmental><drug/agent><efficacy analysis><efficacy assessment><efficacy determination><efficacy evaluation><efficacy examination><efficacy testing><evaluate efficacy><examine efficacy><experience><gene editing method><gene editing methodology><gene editing strategy><gene editing techniques><gene repair therapy><gene therapy><gene-based therapy><gene-based treatment><gene-directed therapy><gene-editing approach><gene-editing therapy><gene-targeted therapy><gene-targeted treatment><genetic therapy><genome editing based therapy><genome editing therapy><genome editing treatment><genome editing-based therapeutics><genome integrity><genomic integrity><genomic therapy><genotoxicity><great ape><hepatic body system><hepatic organ system><hepatic toxicity><hepatitis B virus animal model><hepatoxicity><host response><humanized mice><humanized mouse><immune competent><immune system response><immune therapeutic approach><immune therapeutic interventions><immune therapeutic regimens><immune therapeutic strategy><immune therapy><immune-based therapies><immune-based treatments><immuno therapy><immunogenicity><immunoresponse><improved><in vivo><infected with HBV><infected with hepatitis B><infected with hepatitis B virus><infection with HBV><infection with hepatitis B virus><inhibitor><interest><intervention efficacy><lipid based nanoparticle><lipid nanoparticle><liver cancer><liver carcinoma><liver malignancy><long-term study><longitudinal outcome studies><longterm study><mRNA><malignant liver tumor><model of animal><mouse model><murine model><nCoV vaccine><nCoV-19 vaccine><nCoV19 vaccine><nano particle delivery><nanoparticle delivered><nanoparticle delivery><neuron toxicity><neuronal toxicity><neurotoxicity><new approaches><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutics><new therapy><next generation therapeutics><novel><novel approaches><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel strategies><novel strategy><novel therapeutics><novel therapy><nuclease><pgRNA><promoter><promotor><serum hepatitis><success><targeted drug therapy><targeted drug treatments><targeted endonucleases><targeted nucleases><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapeutic agent development><therapeutic development><therapeutic editing><therapeutic efficacy><therapeutic genome editing><therapy efficacy><vaccine against 2019-nCov><vaccine against COVID-19><vaccine against SARS-CoV-2><vaccine against SARS-coronavirus-2><vaccine against Severe Acute Respiratory Syndrome CoV 2><vaccine against Severe acute respiratory syndrome coronavirus 2><vaccine candidates against SARS-CoV-2><vaccine for novel coronavirus><vaccines preventing COVID><vaccines to prevent COVID><vector><viraemia><viral DNA><viral infection><viral infectious disease treatment><viral multiplication><viral replication><viral sepsis><virus DNA><virus infection><virus multiplication><virus-induced disease><virusemia>