Juan Gallardo Pinera

Program: Molecular Cell Biology

Current advisor: Fumihiko Urano, MD, PhD

Undergraduate university: University of South Florida, 2022

Enrollment year: 2022

Research summary
A One-Time Base Editing Therapy for Neurodegeneration in Wolfram Syndrome

This project seeks to evaluate a one-time therapeutic strategy using base editing to correct a pathogenic WFS1 mutation (W540*) responsible for neurodegeneration in Wolfram Syndrome (WS). WS is a rare monogenic disorder characterized by diabetes, optic atrophy, hearing loss, and early-onset neurodegeneration, caused by loss-of-function mutations in the WFS1 gene.

The project utilizes adenine base editors (ABEs) to correct the W540* nonsense mutation without causing double-stranded breaks. Two major aims guide the study:

Aim 1 (In Vitro): Assess editing precision and phenotypic rescue in iPSC-derived neural progenitor cells (NPCs) and cortical neurons from WS patients. Edited cells will be evaluated for ER stress, ATP production, and viability.

Aim 2 (In Vivo): Test the therapeutic efficacy of dual-AAV9-delivered base editors in W542* mutant mice via intracerebroventricular injection. Behavioral performance and neuronal survival will be measured to assess phenotypic rescue.

Innovation:
The approach avoids drawbacks of CRISPR-Cas9 by not inducing DSBs, preserves native gene regulation, and aims to provide a single-intervention therapy. Novel tools such as virus-like particles (eVLPs) and dual-AAVs are optimized for delivery.

Impact:
This project pioneers a gene-editing strategy for treating neurodegeneration in WS and offers a framework for applying base editing to other neurogenetic diseases involving ER stress, calcium dysregulation, or mitochondrial dysfunction.

Graduate publications