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Principal Investigator: Abigail Benkert
Organization: DUKE UNIVERSITY
Fiscal Year: 2024
Award: $83,392
Funding agency: National Heart Lung and Blood Institute
Project Summary/Abstract
Disruption of mitochondrial oxidative phosphorylation (OXPHOS) is associated with the development of
biochemical alterations that typically affect tissues with a high energy demand, particularly skeletal and cardiac
muscle. An inherited autosomal recessive skeletal myopathy and hypertrophic cardiomyopathy has been linked
to loss of function of a nuclear DNA-encoded mitochondrial protein, due to a frameshift mutation in solute
carrier family 25, member 4 (SLC25A4; c.523delC, p.Q175RfxX38). SLC25A4 encodes the heart-muscle
isoform of the adenine nucleotide translocator-1 (ANT1, SLC25A4), which in the wild-type state is a critical
component of mitochondrial metabolism. Patients with SLC25A4 deficiency display lactic acidosis, persistent
adrenergic activation, and exertional intolerance secondary to both a general skeletal muscle myopathy as well
as a hypertrophic cardiomyopathy. Ultimately, myocardial thickening and cardiac dysfunction progress to end-
stage heart failure necessitating cardiac transplantation.
There are not currently any disease-modifying therapies available for this patient cohort. However, adeno-
associated viral (AAV) mediated gene replacement therapies have emerged as a powerful strategy for disease
modification of inherited monogenic disorders. The long-term goal of our research is to develop a therapeutic
gene replacement strategy to treat SLC25A4 deficiency. The objective of this proposal is to further characterize
the disease phenotype as well as to synthesize and evaluate the efficacy of a recombinant AAV (rAAV) vector
in an in vitro model of patient-derived cell lines and organoid models. The central hypothesis of this
proposal is that AAV-mediated gene replacement can ameliorate the biochemical and functional effects
of SLC25A4 deficiency and can more decisively prevent disease progression. The specific aims of this
proposal are:
1. Characterize the SLC25A4 deficiency phenotype in patient-derived cell lines.
2. Synthesize a recombinant AAV vector for delivery of codon-optimized SLC25A4 cDNA to
skeletal and cardiac myocytes.
3. Evaluate the efficacy of AAV-SLC25A4 viral transduction in patient-derived cell lines.
These experiments will improve our understanding of the molecular mechanisms underlying SLC25A4
deficiency as well as allow us to evaluate the efficacy of an AAV platform in a relevant preclinical model.
Moreover, the skills I will acquire during this fellowship will help to establish me as an independent investigator
and a surgeon-scientist focused on the development of translational gene replacement therapies.
Terms: <ADP-ATP Translocase-1><ADP/ATP Carrier 1><ADP/ATP Translocator of Skeletal Muscle><ANT gene><ANT protein><ANT1><Adenine Nucleotide Translocator 1><Adrenergic Agents><Adrenergic Drugs><Adrenergics><Affect><Asymmetric Septal Hypertrophy><Base Pairing><Biochemical><Body Tissues><Cardiac Muscle Cells><Cardiac Myocytes><Cardiac Transplantation><Cardiocyte><Cardiomyopathies><Cell Body><Cell Line><Cell model><CellLine><Cells><Cellular model><Codon><Codon Nucleotides><Complementary DNA><DNA><Data><Deoxyribonucleic Acid><Development><Disease><Disease Progression><Disorder><Dysfunction><Evaluation><Exertion><Family><Fellowship><Foundations><Frame Shift Mutation><Frameshift Mutation><Functional disorder><Generations><Genetic Alteration><Genetic Change><Genetic defect><Goals><Heart Grafting><Heart Muscle Cells><Heart Transplantation><Heart failure><Heart myocyte><Hereditary><Hereditary Disease><Hereditary ventricular hypertrophy><Hypertrophic Cardiomyopathy><Hypertrophic Obstructive Cardiomyopathy><Idiopathic Hypertrophic Subvalvular Stenosis><Idiopathic hypertrophic subaortic stenosis><Impairment><In Vitro><Inborn Genetic Diseases><Individual><Inherited><Inherited disorder><Inner mitochondrial membrane><Investigators><Isoforms><Laboratories><Lactic Acidosis><Link><Mediating><Mendelian disease><Mendelian disorder><Mendelian genetic disorder><Mennonite><Mitochondria><Mitochondrial Proteins><Modality><Modeling><Modification><Molecular><Muscle Cells><Muscle Disease><Muscle Disorders><Muscle Fibers><Muscular Diseases><Mutation><Myocardial><Myocardial Diseases><Myocardial Disorder><Myocardial depression><Myocardial dysfunction><Myocardiopathies><Myocardium><Myocytes><Myopathic Conditions><Myopathic Diseases and Syndromes><Myopathic disease or syndrome><Myopathy><Myotubes><Natural History><Nuclear><Organ><Organoids><Oxidative Phosphorylation><Oxidative Phosphorylation Pathway><Patients><Pedigree><Phenotype><Physiopathology><Pre-Clinical Model><Preclinical Models><Protein Isoforms><Proteins><Reading Frame Shift Mutation><Recombinants><Recovery><Research><Research Personnel><Researchers><Rhabdomyocyte><SLC25A4><SLC25A4 gene><SYS-TX><Scientist><Secondary to><Skeletal Development><Skeletal Fiber><Skeletal Muscle><Skeletal Muscle Cell><Skeletal Muscle Fiber><Skeletal Myocytes><Solute Carrier Family 25(Mitochondrial Carrier; Adenine Nucleotide Translocator), Member 4><Strains Cell Lines><Structure><Surgeon><Systemic Therapy><Therapeutic><Tissues><Training><Transgenes><Translating><Translational Research><Translational Science><Translations><Tropism><Viral><Viral Vector><Voluntary Muscle><alleviate symptom><ameliorating symptom><autosome><cDNA><cardiac dysfunction><cardiac failure><cardiac graft><cardiac muscle><cardiomyocyte><career><cohort><cultured cell line><decrease symptom><delivery vector><delivery vehicle><determine efficacy><developmental><disease phenotype><efficacy analysis><efficacy assessment><efficacy determination><efficacy evaluation><efficacy examination><evaluate efficacy><examine efficacy><experiment><experimental research><experimental study><experiments><fewer symptoms><gene null><gene replacement><gene replacement therapy><gene therapeutics><gene-based therapeutic><gene-based therapeutics><genes therapeutic><genes therapeutics><genetic pedigree><genome mutation><heart dysfunction><heart muscle><heart transplant><hereditary disorder><heritable disorder><hiPSC><human iPS><human iPSC><human induced pluripotent cell><human induced pluripotent stem cells><human inducible stem cells><hypertrophic myocardiopathy><improved><in vitro Model><inborn error><induced human pluripotent stem cells><induced pluripotent stem cells derived from patients><induced pluripotent stem cells from patients><inherited diseases><inherited genetic disease><inherited genetic disorder><loss of function><member><mitochondrial><mitochondrial metabolism><monogenic disease><monogenic disorder><muscular disorder><myocardium disease><myocardium disorder><next generation><null mutation><pathophysiology><patient derived human iPS><patient derived human iPSC><patient derived human induced pluripotent stem cell><patient derived iPS><patient derived iPSC><patient derived induced pluripotent cells><patient derived induced pluripotent stem cells><patient-derived pluripotent stem cells><pedigree structure><premature><prematurity><prevent><preventing><reduce symptoms><relieves symptoms><restoration><single-gene disease><single-gene disorder><skeletal><skills><solute><symptom alleviation><symptom reduction><symptom relief><therapeutic gene><transgene><translation><translation research><translational investigation><treatment strategy><vector><vector biodistribution>