Document text
Principal Investigator: Eric T Wang
Organization: UNIVERSITY OF FLORIDA
Fiscal Year: 2024
Award: $531,109
Funding agency: National Institute of Arthritis and Musculoskeletal and Skin Diseases
Gene editing is moving towards the clinic, but several challenges must be addressed in order for it to be applied
to skeletal muscle disease. Adeno-associated virus (AAV) is one of the most promising technologies to enable
delivery of gene editing machinery to skeletal muscle, but at least four major hurdles stand in the way of using it
to safely and effectively edit genes in this tissue. These include 1) maximizing delivery of editing enzymes to
muscle, 2) maximizing spread of editing enzymes across nuclei of the myofiber syncytium, 3) avoiding sustained
expression of editing enzymes, and 4) integrating solutions to each of these problems in a single AAV so as to
minimize total viral dose required. Given that exciting new myotropic capsids that have emerged in the literature,
this proposal is primarily focused on addressing the latter 3 issues. We have previously developed ways to
shuttle Cas enzymes throughout myofibers and now propose to use creative new approaches that rely on tried-
and-true mechanisms to access and edit even more myonuclei in skeletal muscle. We have also developed ways
to use small molecules to control AAV-delivered gene expression and propose to further develop this technology
to achieve fully “off” transgene states with maximally “on” states when desired, so that Cas enzyme expression
can be turned off when no longer needed. Finally, we will integrate these technologies with the latest miniaturized
Cas enzymes and package all components, including guide RNAs, into a single AAV vector. We will test
approaches in mouse models of myotonic dystrophy and Duchenne muscular dystrophy. Ultimately, the insights
we make here will be apply to gene editing for any skeletal muscle disease and enable the next generation of
therapies for muscular dystrophies.
Terms: <AAV vector><AAV-based vector><Address><Adeno-Associated Viruses><Alternate Splicing><Alternative RNA Splicing><Alternative Splicing><Architecture><Autograft><Autologous Transplantation><Autotransplant><BCAR1><BCAR1 Protein><BCAR1 gene><Back><Blood Precursor Cell><Body Tissues><Bone Marrow><Bone Marrow Reticuloendothelial System><Breast Cancer Anti-Estrogen Resistance 1 Protein><CKRAS protein><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><CRK-Associated Substrate><CRKAS><Capsid><Cas nuclease technology><Cas protein><Cell Body><Cell Nucleus><Cell Nucleus Active Transport><Cells><Climacteric><Clinic><Clinical Trials><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><Creativeness><Cytoplasm><DNA cassette><Dependoparvovirus><Dependovirus><Disease><Disorder><Dorsum><Dose><Drugs><Duchene><Duchenne><Duchenne muscular dystrophy><Duchenne-Griesinger syndrome><Dystrophia Myotonica><Dystrophia myotonica type 1><Elements><Ellis-van Creveld (EvC) syndrome><Engineering / Architecture><Enzyme Gene><Enzymes><Essential Hypercholesterolemia><Estrogen Receptors><FDA approved><Facioscapulohumeral><Familial Hypercholesterolemia><Gene Delivery><Gene Expression><Gene Therapy Vectors><Gene Transduction Agent><Gene Transduction Vectors><Generations><Genes><Giant Cells><Goals><Guide RNA><Hb SS disease><HbSS disease><Hematopoietic Progenitor Cells><Hematopoietic stem cells><Hemoglobin S Disease><Hemoglobin sickle cell disease><Hemoglobin sickle cell disorder><Hyperbetalipoproteinemia><Hyperlipoproteinemia Type 2><Hyperlipoproteinemia Type II><Image><Immune Cell Activation><Immune response><Immunological response><Individual><LDL><LDL Lipoproteins><Limb-Girdle Muscular Dystrophies><Literature><Liver><Low-Density Lipoproteins><Medication><Messenger RNA><Methods><Micro-tubule><Microtubules><Modality><Multinucleated Giant Cells><Muscle><Muscle Disease><Muscle Disorders><Muscle Tissue><Muscular Diseases><Muscular Dystrophies><Myodystrophica><Myodystrophy><Myopathic Conditions><Myopathic Diseases and Syndromes><Myopathic disease or syndrome><Myopathy><Myotonia Atrophica><Myotonia Dystrophica><Myotonic Dystrophy><Myotonic dystrophy type 1><Non-Polyadenylated RNA><Nuclear><Nuclear Export><Nuclear Import><Nuclear Transport><Nucleocytoplasmic Shuttling><Nucleus><Patients><Peptides><Pharmaceutical Preparations><Polykaryocytes><Proteins><Pseudohypertrophic Muscular Dystrophy><Public Health><Quality Control><RNA><RNA Gene Products><RNA Splicing><RNA-Binding Proteins><Regulation><Research><Ribonucleic Acid><Route><Safety><Science><Sickle Cell Anemia><Skeletal Muscle><Speed><Splicing><Steinert Disease><Syncytium><System><Technology><Testing><Time><Tissues><Transgenes><Type 2 Hyperlipidemia><Type II Hyperlipidemia><Tzanck Cell><Viral><Voluntary Muscle><Work><X-linked dilated cardiomyopathy><X-linked muscular dystrophy><X-linked recessive muscular dystrophy><adeno associated virus group><adeno-associated viral vector><adeno-associated virus vector><antibody conjugate><autologous graft><autotransplantation><base editing><base editor><behavior test><behavioral test><benign X-linked recessive muscular dystrophy><beta-Lipoproteins><blood cell progenitor><blood progenitor><blood stem cell><blood-forming stem cell><cell type><childhood pseudohypertrophic muscular dystrophy><classic X-linked recessive muscular dystrophy><creativity><debilitating pain><deliver mRNA><deliver messenger RNA><delivery system for mRNA><design><designing><drug/agent><dystrophic myotonia><enhancer cassette><experience><expression cassette><familial hyperbetalipoproteinemia><familial hypercholesteremia><familial hyperlipoproteinemia type 2><familial hyperlipoproteinemia type II><fiction><fictional works><gRNA><gene cassette><gene editing method><gene editing methodology><gene editing platform><gene editing strategy><gene editing system><gene editing techniques><gene editing technology><gene editing tools><gene-editing approach><gene-editing toolkit><genetic cassette><genome editing><genomic editing><hematopoietic progenitor><hematopoietic stem progenitor cell><hemopoietic progenitor><hemopoietic stem cell><hepatic body system><hepatic organ system><host response><imaging><immune activation><immune system response><immunoresponse><improved><in vivo><insight><integration cassette><life change><limb-girdle muscular weakness and atrophy><limb-girdle syndrome><lipid based nanoparticle><lipid nanoparticle><mRNA><mRNA delivery><messenger RNA delivery><mild X-linked recessive muscular dystrophy><miniaturize><miniaturized><mouse model><murine model><muscle dystrophy><muscle progenitor cell><muscle stem cell><muscular><muscular disorder><muscular dystrophy mouse model><myopathic limb-girdle syndrome><new approaches><new technology><next generation><novel approaches><novel strategies><novel strategy><novel technologies><nucleocytoplasmic transport><p130 cas protein><p130CAS><progenitor><progressive muscular dystrophy of childhood><promoter cassette><protein expression><pseudohypertrophic adult muscular dystrophy><pseudohypertrophic muscular paralysis><reporter cassette><resistance cassette><screening><screenings><selectable cassette><selection cassette><sickle cell disease><sickle cell disorder><sickle disease><sicklemia><single molecule><skeletal><small molecule><stop cassette><transcription cassette><transcriptional cassette><transgene><transgene cassette><transgene expression><translational opportunities><translational potential>