We recently developed a technology that combines catalytically inactive dCas9 with bifunctional chemicals that link endogenous host cellular machinery to specific genes targeted with dCas9. We have demonstrated this technology can significantly increase gene expression at multiple endogenous mammalian genes in a chemically dependent manner. Here we propose to apply this technology to the mono-allelic rare disease with no current cure, Friedreichs Ataxia. This disease is caused by a GAA expansion in the first intron of the frataxin gene which leads to polymerase pausing and lower gene expression. This proposed project will evaluate if targeting the frataxin gene with our combination of protein and chemical activator can restore expression levels of frataxin to a point which reverses pathology of this disease in a modern model of Friedreichs Ataxia. We will use patient derived Friedreichs Ataxia models in specific assays that intricately measure mitochondria function and metabolic activity that is typically dysregulated in patients that is the hallmark of this human disease. This work has the potential to lead to new treatment paradigms for individuals afflicted by this aggressive disease.
Public Health Relevance Statement: Friedreichs Ataxia affects roughly 1:50,000 individuals is a devastating disease driven by gene dysregulation. The root of the disease is a genetic disruption that causes pausing of transcriptional machinery on the frataxin gene. Here we evaluate the concept of restating paused transcriptional activity with a gene specific chemical activator we have developed in order to alleviate the pathology of this disabling human disease.
Project Terms: Aberrant DNA Methylation; Acetylation; Affect; Alleles; Arrhythmia; Ataxia; Automobile Driving; base; Binding; Bioenergetics; Biological Assay; Cardiac; Cardiomyopathies; Cell model; Cells; Chemicals; Chimeric Proteins; Chromatin; chromatin modification; Clinical; clinical translation; clinically relevant; Confocal Microscopy; Coupled; CRISPR/Cas technology; curative treatments; Development; Diabetes Mellitus; Disease; disease phenotype; DNA Sequence; Engineering; Enzymes; Epigenetic Process; epigenetic regulation; epigenome; experimental study; Expression Profiling; extracellular; FDA approved; Fibroblasts; FK506; frataxin; Friedreich Ataxia; Functional disorder; Galactose; Gene Activation; Gene Expression; Gene Proteins; gene repression; Gene Targeting; Generations; Genes; Genetic; Genetic Transcription; Genome; Genomics; Genus Hippocampus; Guide RNA; Histone Deacetylase; Histone Deacetylase Inhibitor; Histone H3; Histones; Hour; human disease; Individual; inhibitor/antagonist; innovation; Introns; Iron; Lead; Link; Measures; Mediating; Metabolic; Metabolism; Methylation; Mitochondria; mitochondrial dysfunction; Mitochondrial Proteins; Modeling; Modernization; mRNA Expression; Myasthenia; Neurodegenerative Disorders; Neurologic; novel; Nuclear; Oxygen Consumption; Pathology; Pathway interactions; Patients; Peroxidases; Pharmaceutical Preparations; Plant Roots; Polyethylene Glycols; Polymerase; protein expression; Proteins; Rare Diseases; rare variant; Respiration; RNA delivery; scoliosis; Signal Pathway; Site; Stress; System; Tacrolimus Binding Proteins; Technology; Therapeutic; therapeutic gene; Transcription Initiation Site; vector; Virginia; Work
