Development of a Cellular System to Screen for Small Molecules that Upregulate the hVMAT2 Promoter Activity
Rapid Response Innovation Awards, 2007
We will develop an essential cellular system as a platform for future high throughput screen (HTS) for novel small molecules that increase levels of the human vesicular monoamine transporter 2 gene (hVMAT2). The function of the hVMAT2 protein is to sequester cytosolic toxins into intracelluar vesicles, preventing neuron death which is the cause of PD. Clinical and pre-clinical studies have shown that lower hVMAT2 expression is associated with PD and enhanced hVMAT2 expression displays neuroprotection against PD. Identification of hVMAT2-upregulating small molecules may lead to medications development.
To establish the cellular system, we will genetically engineer SH-SY5Y cells to link the expression of hVMAT2 with a reporter gene that can easily be visualized. In this way, we will be able to easily detect expression levels of hVMAT2. We will utilize cell cloning techniques to prepare a large amount of uniform cells that can be used in a high throughput assay. Specifically, this project contains three steps: 1) genetic engineering of hVMAT2, 2) generation of SH-SY5Y cell lines stably expressing the genes of interest and 3) characterization and selection of the stable cells suitable for HTS.
Relevance to Diagnosis/Treatment of Parkinson’s Disease:
Establishment of this cellular system will facilitate HTS for small molecules that upregulate hVMAT2 expression and confer neuroprotection, leading to development of novel preventive and therapeutic medications for PD.
Three to five SH-SY5Y cell lines will be obtained for use in future HTS.
Dr. Lin generated an assay for screening compounds that may upregulate hVMAT2 promoter activity. Additional work however, is needed to determine whether this assay could be used for high throughput screening of compounds.
Instructor in Psychiatry at McLean Hospital (Harvard Medical School)
Location: Boston, Massachusetts