Using human pluripotent stem cell-derived dopaminergic neurons to evaluate candidate Parkinson's disease therapeutic agents in MPP+ and rotenone models.
To begin to develop a high-throughput assay system to evaluate potential small-molecule therapy for Parkinson's disease (PD), we have performed a low-throughput assay with a small number of compounds using human pluripotent stem cell-derived dopaminergic neurons. We first evaluated the role of 44 compounds known to work in rodent systems in a 1-methyl-4-phenylpyridinium (MPP(+)) assay in a 96-well format using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay as a readout for neuroprotection. Glial cell-derived neurotrophic factor was used as a positive control because of its well-documented neuroprotective effect on dopaminergic neurons, and two concentrations of each drug were tested. Of 44 compounds screened, 16 showed a neuroprotective effect at one or both dosages tested. A dose-response curve of a subset of the 16 positives was established in the MPP(+) model. In addition, we validated neuroprotective effects of these compounds in a rotenone-induced dopaminergic neuronal cell death, another established model for PD. Our human primary dopaminergic neuron-based assays provide a platform for rapid screening and/or validation of potential neuroprotective agents in PD treatment using patient-specific cells and show the importance of using human cells for such assays.
Buck Institute for Research on Aging, Novato, CA, USA.
Jun
J
Peng
Qiuyue
Q
Liu
Mahendra S
MS
Rao
Xianmin
X
Zeng
D015655:1-Methyl-4-phenylpyridinium; D000978:Antiparkinson Agents / Q000302:isolation & purification* / Q000627:therapeutic use; D002454:Cell Differentiation; D002478:Cells, Cultured; D059290:Dopaminergic Neurons / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D057166:High-Throughput Screening Assays; D006801:Humans; D046228:Microarray Analysis; D008962:Models, Theoretical; D055495:Neurogenesis / Q000502:physiology; D018696:Neuroprotective Agents / Q000302:isolation & purification / Q000627:therapeutic use; D010302:Parkinson Disease, Secondary / Q000139:chemically induced / Q000188:drug therapy* / Q000473:pathology; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology*; D012402:Rotenone / Q000633:toxicity
D000978:Antiparkinson Agents; D018696:Neuroprotective Agents; D012402:Rotenone; D015655:1-Methyl-4-phenylpyridinium
dopaminergic neurons; neuroprotective; parkinsonism; stem cells