[HTML][HTML] Efficient conversion of astrocytes to functional midbrain dopaminergic neurons using a single polycistronic vector

RC Addis, FC Hsu, RL Wright, MA Dichter, DA Coulter… - PloS one, 2011 - journals.plos.org
RC Addis, FC Hsu, RL Wright, MA Dichter, DA Coulter, JD Gearhart
PloS one, 2011journals.plos.org
Direct cellular reprogramming is a powerful new tool for regenerative medicine. In efforts to
understand and treat Parkinson's Disease (PD), which is marked by the degeneration of
dopaminergic neurons in the midbrain, direct reprogramming provides a valuable new
source of these cells. Astrocytes, the most plentiful cells in the central nervous system, are
an ideal starting population for the direct generation of dopaminergic neurons. In addition to
their potential utility in cell replacement therapies for PD or in modeling the disease in vitro …
Direct cellular reprogramming is a powerful new tool for regenerative medicine. In efforts to understand and treat Parkinson's Disease (PD), which is marked by the degeneration of dopaminergic neurons in the midbrain, direct reprogramming provides a valuable new source of these cells. Astrocytes, the most plentiful cells in the central nervous system, are an ideal starting population for the direct generation of dopaminergic neurons. In addition to their potential utility in cell replacement therapies for PD or in modeling the disease in vitro, astrocyte-derived dopaminergic neurons offer the prospect of direct in vivo reprogramming within the brain. As a first step toward this goal, we report the reprogramming of astrocytes to dopaminergic neurons using three transcription factors – ASCL1, LMX1B, and NURR1 – delivered in a single polycistronic lentiviral vector. The process is efficient, with 18.2±1.5% of cells expressing markers of dopaminergic neurons after two weeks. The neurons exhibit expression profiles and electrophysiological characteristics consistent with midbrain dopaminergic neurons, notably including spontaneous pacemaking activity, stimulated release of dopamine, and calcium oscillations. The present study is the first demonstration that a single vector can mediate reprogramming to dopaminergic neurons, and indicates that astrocytes are an ideal starting population for the direct generation of dopaminergic neurons.
PLOS