title: Viral Vector Reprogramming of Adult Resident Striatal Oligodendrocytes into Functional Neurons.
abstract: Recent advances suggest that in vivo reprogramming of endogenous cell populations provides a viable alternative for neuron replacement. Astrocytes and oligodendrocyte precursor cells can be induced to transdifferentiate into neurons in the CNS, but, in these instances, reprogramming requires either transgenic mice or retroviral-mediated gene expression. We developed a microRNA (miRNA)-GFP construct that in vitro significantly reduced the expression of polypyrimidine tract-binding protein, and, subsequently, we packaged this construct in a novel oligodendrocyte preferring adeno-associated virus vector. Ten days after rat striatal transduction, the vast majority of the GFP-positive cells were oligodendrocytes, but 6 weeks to 6 months later, the majority of GFP-positive cells exhibited neuronal morphology and co-localized with the neuronal marker NeuN. Patch-clamp studies on striatal slices established that the GFP-positive cells exhibited electrophysiological properties indicative of mature neurons, such as spontaneous action potentials and spontaneous inhibitory postsynaptic currents. Also, 3 months after striatal vector administration, GFP-positive terminals in the ipsilateral globus pallidus or substantia nigra retrogradely transported fluorescent beads back to GFP-positive striatal cell bodies, indicating the presence of functional presynaptic terminals. Thus, this viral vector approach provides a potential means to harness resident oligodendrocytes as an endogenous source for in vivo neuronal replacement.
authors.affiliation: UNC Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
authors.forename: Marc S
authors.initials: MS
authors.lastname: Weinberg
authors.affiliation: Bowles Center for Alcohol Studies, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Department of Psychiatry, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
authors.forename: Hugh E
authors.initials: HE
authors.lastname: Criswell
authors.affiliation: UNC Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
authors.forename: Sara K
authors.initials: SK
authors.lastname: Powell
authors.affiliation: Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
authors.forename: Aadra P
authors.initials: AP
authors.lastname: Bhatt
authors.affiliation: UNC Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Department of Psychiatry, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA. Electronic address: thomas_mccown@med.unc.edu.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: McCown
mesh_terms: D000818:Animals; D002460:Cell Line; D054338:Cell Transdifferentiation / Q000235:genetics*; D065150:Cellular Reprogramming / Q000235:genetics*; D003342:Corpus Striatum / Q000166:cytology*; D000229:Dependovirus / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology* / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering; D051381:Rats
chemical_list: D034741:RNA, Small Interfering
keywords: adeno-associated virus; gene therapy; neurons; oligodendrocytes; reprogramming; transdifferentiation