Study Rationale: This project represents a supplement to our MJFF project, “Shedding Light on the Dark Genome in Parkinson’s Disease (PD).” For the initial project, we developed a suite of standardized cellular assays to evaluate key PD traits. Based on these achievements and the potential of this approach, we will now expand our screening capabilities in induced-pluripotent stem cells (iPSC), with the goal of establishing a robust platform to study the mechanisms of action and potential pathological role of virtually any PD-related gene.
Hypothesis: We hypothesize that optimizing our existing assays, and adding new ones, will allow us to capture key phenotypes and mechanisms of PD pathology and will provide a robust discovery and validation pipeline for investigating a broader range of PD-related genes.
Study Design: For this project, we will optimize, expand and validate our PD-relevant cellular assays based on human iPSC. We will transition to using a newer method (based on CRISPR activation and interference) to control the activity of potential PD-associated genes in our iPSC systems. We will characterize and compare features of cells with PD-associated mutations (derived directly from people with PD) with those in which the mutation is voluntarily introduced in a healthy cell line. Finally, we will expand our repertoire of PD-relevant cell types beyond neurons by generating proof-of-concept preliminary data using microglia and astrocytes.
Impact on Diagnosis/Treatment of Parkinson’s disease:
This platform will act as a strong discovery and validation pipeline to explore a wider array of targets related to PD, providing deeper understanding of the disease's underlying mechanisms.
Next Steps for Development: Next steps will involve adding new PD genes as well as incorporating other brain cell types, such as oligodendroglia, relevant for multiple systems atrophy; we will also use iPSC-derived midbrain organoids, which contain multiple cell types growing in 3D, to explore the different cellular pathways associated with PD pathogenesis.