Integrating Environmental, Genomic, and Functional Data to Characterize Individual Risk for Parkinson's Disease
Abstract
The overall purpose of this work is to identify methods for characterizing which individuals may be particularly susceptible to specific toxic insults, and the underlying mechanisms. We have three Aims: 1) use iPSC-derived neuronal cell lines to validate and extend our prior work that found an 11-fold increased risk of Parkinson's disease (PD) in persons exposed to the herbicide paraquat who lacked functional glutathione-S-transferase T1 (GSTT1), a key metabolic enzyme with anti-oxidant function; 2) perform whole genome sequencing of existing DNA specimens from a PD study with highly characterized pesticide exposures, using bioinformatic tools to identify likely functional gene-environment interactions; and 3) test our predictions using iPSC-derived neuronal cell lines that model these genetic variations, and exposing cellular lines to these same environmental agents. This is a partnering PI project with W81XWH-20-1-0709. The scope of the -0710 project is to accomplish Aims 1 and 3. In work performed for Aim 1 during this reporting period, we established 3 clones of iPSC-derived dopaminergic neurons and have down-regulated GSTT1 in one of these lines so far. We have also established markers of oxidative stress and cell death, and used these markers to established dose-response curves for the 6 toxicants pertinent to this study. For superoxide formation, DNA damage, and cell death. We have also established long-term iPSC cultures for assessing -synuclein aggregate formation, induced by viral-mediated -syn overexpression and exposure to pre-formed -syn fibrils. Work ongoing is evaluating the effect of CRISPRi - mediated GTT1 downregulation on these dose-response curves, in cells with and without alpha-synuclein aggregates. In the next reporting period, selected genes identified in the Aim 2 work (W81XWH-20-1-0709) will be downregulated using this same CRISPRi method to assess for biological importance.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 2022
- Accession Number
- AD1190942
Entities
People
- Raymond A. Swanson
Organizations
- Northern California Institute for Research and Education