The development of neuroprotective strategies to prevent neuronal cell death in Parkinson's disease would be enhanced by genetic cell models. Screening of such a cell model with small molecules could identify lead candidates for drug development. Expression of mutant proteins in culture can be used to model autosomal dominant neurodegenerative diseases. We have developed an inducible cell model expression of mutant huntingtin in PC12 cells. We have shown that this model can be used for medium throughput screens by testing the NINDS compound collection of 1040 FDA approved compounds. The hit rate in this screen was approximately 0.4 - 0.5%. We now propose to develop a similar genetic cell model for Parkinson's disease, using inducible expression of mutant alpha-synuclein. We have preliminary data indicating that over expression E46K mutant alpha-synuclein itself causes neuronal cell death in differentiated PC12 cells. We will test current candidate neuroprotective agents, and perform a screen of the NINDS compound collection to demonstrate the utility for moderate throughput screens.
In Specific Aim 1, we will characterize several PC12 cell lines inducibly expressing E46K alpha-synuclein, in order to obtain lines with low background expression, high inducible expression, and consistent toxicity. We will optimize conditions and get a reliable assay for screening.
In Specific Aim 2, we will test compounds currently considered candidates for neuroprotective therapy for PD, such as Co-enzyme Q10, caspase inhibitor z-VAD and others to determine whether they have neuroprotective activity in our model.
In Specific Aim 3, we will carry out a medium throughput screen of the 1040 NINDS compound collection of FDA-approved and related compounds, for inhibition of toxicity. All positives will be confirmed, and dose- response curve will be established to determine approximate inhibitory potencies. Validated positive hits could then be taken to testing in PD animal models in future.Cell Model of Parkinson's Disease and Screening for Therapeutic Compounds ? ? Project Narrative: Parkinson's disease (PD) is the second most common neurodegenerative disease. There is currently no cure or effective neuroprotective treatment. Our long-term goal is to develop effective neuroprotective agents in PD models in order to prepare for clinical therapeutic trials in PD. Our proposed research has potential to find compounds which are neuroprotective and further be taken to preclinical and clinical trials of PD. ? ? ?
| Jiang, Mali; Porat-Shliom, Yair; Pei, Zhong et al. (2010) Baicalein reduces E46K alpha-synuclein aggregation in vitro and protects cells against E46K alpha-synuclein toxicity in cell models of familiar Parkinsonism. J Neurochem 114:419-29 |
