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Abstract Details

Transfer of Therapeutic miRNAs Within Extracellular Vesicles Secreted from Huntington’s Disease iPSC-derived Neurons
Movement Disorders
P5 - Poster Session 5 (8:00 AM-9:00 AM)
3-001

To investigate presence of microRNA targeting human huntingtin (miHTT) molecules within extracellular vesicles (EVs) secreted from adeno-associated serotype 5 (AAV5)-miHTT-treated neuronal cultures from a Huntington’s disease (HD) patient.

HD is a neurodegenerative disorder caused by an autosomal dominant mutation in the huntingtin gene (HTT), leading to mutant protein aggregation, toxicity and neuronal cell death. microRNA (miRNA)-based gene therapies are a potential approach to reduce disease-causing protein in HD. We developed an engineered miHTT, delivered via AAV5 virus. AAV5-miHTT have demonstrated an efficient lowering in vitro and in vivo in the brain in different HD animal models. For efficacy in HD, brain-wide biodistribution (striatum, cortex) of the therapeutic miRNA is of crucial importance. Recently, EVs, including exosomes and microvesicles, have been identified as carriers of RNA species. Understanding how therapeutic miRNAs transfer between neuronal cells is relevant for delivery, translational studies, and biomarker discovery in gene therapies for brain disorders.

An in vitro model based on the differentiation of induced pluripotent stem cells (iPSCs) from HD patients to mature neuronal cells was developed. iPSC-derived neuronal cultures were transduced with AAV5-miHTT and the cultured medium collected for precipitation of EVs.
Dose-dependent transduction of neuronal cells and cellular expression of miHTT was observed as measured by TaqMan qPCR.  Five and 12 days post-transduction, mature miHTT molecules within EVs enriched for exosomes and microvesicles secreted from AAV-transduced iPSC-derived neurons were detected. EV-associated miHTT levels correlated with AAV dose and miHTT expression in neuronal cells. Isolated EVs were taken up by other naïve neuronal cells resulting in a concentration-dependent transfer of miHTT molecules to recipient cells.
EVs contribute to the transfer of therapeutic molecules for miRNA-based gene therapies for HD. EV-containing therapeutic miRNAs can be regarded as novel mediators of cell-cell communication and as a potential source of biomarkers in neurodegenerative disorders.
Authors/Disclosures

PRESENTER
No disclosure on file
No disclosure on file
No disclosure on file
No disclosure on file
No disclosure on file
Melvin Evers No disclosure on file