Pharmacochaperoning in a Drosophila model system rescues human dopamine transporter variants associated with infantile/juvenile parkinsonism.

摘要: Point mutations in the gene encoding human dopamine transporter (hDAT, SLC6A3) cause a syndrome of infantile/juvenile dystonia and parkinsonism. To unravel molecular mechanism underlying these disorders investigate possible pharmacological therapies, here we examined 13 disease-causing DAT mutants that were retained endoplasmic reticulum when heterologously expressed HEK293 cells. In three mutants, i.e. hDAT-V158F, hDAT-G327R, hDAT-L368Q, folding deficit was remedied with pharmacochaperone noribogaine or heat shock protein 70 (HSP70) inhibitor pifithrin-μ such export radioligand binding substrate uptake by restored. Drosophila melanogaster, deficiency results reduced sleep. We therefore exploited power targeted transgene expression mutant hDAT to explore whether could also be pharmacologically rescued an intact organism. Noribogaine treatment supported delivery presynaptic terminals dopaminergic neurons restored sleep normal length DAT-deficient (fumin) lines expressing hDAT-V158F hDAT-G327R. contrast, hDAT-L368Q background caused developmental lethality, indicating toxic action not pharmacochaperoning. Our observations identified those most likely amenable rescue affected children. addition, our findings highlight challenges translating insights from pharmacochaperoning cell culture clinical situation. Because evolutionary conservation neurotransmission between people, D. melanogaster may allow us bridge gap.