Stem Cell-Based Approaches to Identify New Drugs for Treating Dry AMD

The goal of this study is to use state-of-the-art human stem cell RPE models for high-content screening approaches to identify novel small molecules and molecular pathways that inhibit Retinal Pigment Epithelial (RPE) epithelial to mesenchymal transition (EMT) and promote RPE survival in AMD.

We will develop a high-content assay to identify small molecule inhibitors of RPE-EMT and drugs that promote RPE survival against AMD-relevant stressors including cigarette smoke extract and A2E (a toxic byproduct of the visual cycle) in combination with exposure to blue light. Next, we will use gain-of-function (GOF) and loss-of-function (LOF) studies to modulate key components of biological pathways that are implicated in RPE-EMT and RPE cell death. We developed both an inducible CRISPRi (interference) and CRISPRa (activation) stem cell line which we will use in these studies.

Innovative aspects of our proposal include the combined use of stem cell-derived RPE with CRISPRi/a-based genome editing methodology to generate a platform in which we can readily assess the engagement of the RPE-EMT while simultaneously monitoring the “RPE-ness” of the same cell while either modulating gene expression of candidate regulators of EMT and testing compounds for their effect on RPE cell death.

Not only do we benefit from the potential to identify compounds or genetic pathways that can be developed for novel AMD therapeutic applications, but more broadly we envision this platform, the combination of stem-cell-derived human disease models with the power to perform high-content screening and gain and loss of gene-function studies, as a valuable resource for the AMD research community. In the future, we hope to expand panels of RPE differentiated from patients with different AMD-risk associated SNPs to understand environmental and genetic interaction that facilitate individualized AMD.