Funding Summary
Dr. Lu will use an innovative delivery system to achieve CRISPR gene activation in a new mouse model of PWS.
Dr. Theresa Strong, Director of Research Programs, shares details on this project in this short video clip.
Lay Abstract
Recent advances in genetic technology, specifically CRISPR-based epigenome editing, offer a new approach to potentially treat PWS by reactivating the paternally lost genes from maternal chromosome. Our study aims to develop and optimize a new gene therapy method using a non-viral delivery system called STEP-RNP-dCas9/sgRNA to target and reactivate silenced genes in PWS. The project has three main aims: Aim 1: Test the effectiveness of delivering two different epigenetic tools, STEP-RNP-dCas9-JMJD2a/sgRNA and STEP-RNP-dCas9-TET1/sgRNA, in a mouse model that carries a reporter gene to measure reactivation. Our preliminary studies showed that these tools can successfully reactivate genes when injected into juvenile and young adult mice, improving their PWS symptoms. We will refine this method to determine the best way to deliver these tools, either through intravenous (IV) or intrathecal (IT) injection. Aim 2: Investigate whether this gene therapy can correct symptoms in a newly developed PWS mouse model that closely mimics more than 70% PWS patients. This mouse model has a large deletion on chromosome 15, spanning 6 Mb, similar to what is seen in the majority of PWS patients. We will test if our epigenetic tools can reverse the symptoms and reactivate the necessary genes. Additionally, we will compare the effectiveness of these two epigenetic tools at different stages of development (newborn, childhood, and adult). Aim 3: Transition our gene therapy approach from mouse models to human cells, which is crucial for future clinical applications. We will test human-specific versions of our epigenetic tools in cells derived from PWS patients. Specifically, we will use induced pluripotent stem cells (iPSCs) and neurons derived from these iPSCs to see if we can reactivate the silenced genes in human cells. We will also evaluate any unintended effects of our treatment.
Preliminary promising results shown that our STEP-RNP-dCas9/sgRNA system can effectively reactivate silenced genes in mouse and human cells. This research will help refine this epigenetic therapy approach, making it more effective and safer. Ultimately, our goal is to develop a treatment that can significantly improve the quality of life for individuals with PWS.
Funded Year:
2024
Awarded to:
Xiaona Lu, Ph.D.
Amount:
$162,000
Institution:
Yale University School of Medicine
Researcher:
Xiaona Lu, Ph.D.
