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Principal Investigator: Gregory A. Newby
Organization: JOHNS HOPKINS UNIVERSITY
Fiscal Year: 2024
Award: $249,000
Funding agency: National Heart Lung and Blood Institute
PROJECT SUMMARY
Genetic diseases impact over 1 in 50 newborns worldwide and yet there are no approved therapies
capable of correcting the underlying genetic defects. As a result, most patients continue to suffer throughout
their life and require frequent interventions to ameliorate symptoms. I aim to develop in vivo genome editing
therapeutics that correct the underlying disease mutation in relevant tissues by a single injection into the
patient. Base editors can efficiently correct transition point mutations, the most common form of disease-
causing genetic mutation, without undesired editing outcomes such as indels. With one dose and no
subsequent enrichment, over 95% of cells in tissue culture can be edited, and editing in over 60% of non-
dividing cells in targeted adult mammalian tissues has been demonstrated in early in vivo work. Prime editors
can correct any genetic perturbation of up to at least ~50 nt in length (encompassing ~89% of human disease
mutations). I will develop and assess both precision genome editing technologies (base and prime editors)
using suitable in vivo delivery tools in mouse models to develop therapeutics for genetic disease.
Dilated cardiomyopathy (DCM) is a frequent form of genetic heart disease, affecting an estimated
300,000 people in the United States, and can lead to heart failure. Common causes of DCM are
haploinsufficiency of important genes in cardiomyocytes including TTN and LMNA. I will employ screens to
identify new editing strategies to treat haploinsufficiencies by enhancing transcription of the healthy allele. I will
characterize the mechanism of identified edits to understand the associated changes in transcription factor
occupancy and chromatin states. Simultaneously, I will use fluorescent reporter mice to characterize the in vivo
delivery of base editor and prime editor tools in order to find the best method for editing cardiomyocytes. This
work will include characterizations of tissue- and cell-specific editing following delivery via adeno-associated
virus, lipid nanoparticles, or polymer nanoparticles. I will then combine the identified therapeutic editing
strategy with the best vehicle for delivery to cardiomyocytes to treat a mouse model of TTN haploinsufficiency.
I will measure on- and off-target editing as well as any improvement in the contractility defect that defines this
model. Base editors and prime editors can be readily reprogrammed to correct one or even multiple
simultaneous mutations by altering the co-delivered guide RNA. Future work will expand this screening
methodology to additional haploinsufficiency disorders, and applying identified delivery methods to new models
of genetic disease. The ultimate goal of this work is to develop in vivo genome editing therapeutics that can be
readily adapted to treat even rare or one-of-a-kind disease variants.
Terms: <0-4 weeks old><21+ years old><ATAC sequencing><ATAC-seq><ATACseq><Adeno-Associated Viruses><Adult><Adult Human><Affect><Alleles><Allelomorphs><Animal Model><Animal Models and Related Studies><Assay for Transposase-Accessible Chromatin using sequencing><Basal Transcription Factor><Basal transcription factor genes><Binding><Binding Sites><Body Tissues><Brain><Brain Nervous System><Cardiac Diseases><Cardiac Disorders><Cardiac Muscle Cells><Cardiac Myocytes><Cardiocyte><Cell Body><Cell division><Cells><ChIP Sequencing><ChIP-seq><ChIPseq><Chromatin><Clinical><Clinical Trials><Combining Site><Congestive Cardiomyopathy><DNA Alteration><DNA Sequence Alteration><DNA editor><DNA mutation><Data Bases><Databases><Defect><Dependoparvovirus><Dependovirus><Detection><Dilated Cardiomyopathy><Disease><Disorder><Dose><Echocardiogram><Echocardiography><Encephalon><Engineering><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Frequencies><Future><Gene Transcription><General Transcription Factor Gene><General Transcription Factors><Genes><Genetic><Genetic Alteration><Genetic Change><Genetic Diseases><Genetic Models><Genetic Transcription><Genetic defect><Genetic mutation><Goals><Guide RNA><Heart><Heart Diseases><Heart Muscle Cells><Heart failure><Heart myocyte><Human><Injectable><Injections><Interphase Cell><Intervention><Intervention Strategies><Intravenous><Length><Life><Liver><Measures><Mentors><Methodology><Methods><Mice><Mice Mammals><Microscopy><Modality><Modeling><Modern Man><Molecular Interaction><Murine><Mus><Muscle><Muscle Tissue><Mutation><Myocardium><Newborn Infant><Newborns><Non-dividing Cell><Nondividing Cell><Nucleotides><Orphan Disease><Other Genetics><Outcome><Patients><Persons><Phase><Phenotype><Photography><Point Mutation><Population><Postdoc><Postdoctoral Fellow><Promoter Regions><Promotor Regions><RNA Expression><Rare Diseases><Rare Disorder><Reactive Site><Reporter><Research Associate><Resting Cell><Sequence Alteration><Site><Specificity><Technology><Therapeutic><Therapeutic Gene Editing><Tissues><Transcription><Transcription Factor Proto-Oncogene><Transcription factor genes><Transthoracic Echocardiography><United States><Up-Regulation><Upregulation><Variant><Variation><Viral><Work><adeno associated virus group><adulthood><alleviate symptom><ameliorating symptom><assay for transposase accessible chromatin followed by sequencing><assay for transposase accessible chromatin seq><assay for transposase accessible chromatin sequencing><assay for transposase-accessible chromatin with sequencing><base><base editing><base editor><bases><cardiac failure><cardiac muscle><cardiomyocyte><cell type><chromatin immunoprecipitation-sequencing><data base><decrease symptom><delivery vector><delivery vehicle><fewer symptoms><flow cytophotometry><gRNA><gene editor><gene-editing therapy><genetic condition><genetic disorder><genetic promoter element><genetic promoter sequence><genome editing><genome editing based therapy><genome editing therapy><genome editing treatment><genome editing-based therapeutics><genome editor><genome mutation><genomic alteration><genomic editing><heart disorder><heart muscle><heart sonography><hepatic body system><hepatic organ system><human disease><iPS><iPSC><iPSCs><improved><in vivo><indel><induced pluripotent cell><induced pluripotent stem cell><inducible pluripotent stem cell><insertion-deletion><insertion-deletion mutation><insertion/deletion><insertion/deletion mutation><interest><interventional strategy><lipid based nanoparticle><lipid nanoparticle><model of animal><mouse genome><mouse model><murine model><muscular><mutation correction><nano particle><nano particle delivery><nano polymer><nano-sized particle><nanoparticle><nanoparticle delivered><nanoparticle delivery><nanopolymer><nanosized particle><newborn child><newborn children><orphan disorder><post-doc><post-doctoral><post-doctoral trainee><precise genome editing><prime editing><prime editor><promoter><promoter sequence><promotor><reduce symptoms><relieves symptoms><repair><repair strategy><repaired><research associates><screening><screenings><success><symptom alleviation><symptom reduction><symptom relief><therapeutic agent development><therapeutic development><therapeutic editing><therapeutic genome editing><tissue culture><tool><transcription factor>