Oscar Perez-Leal, MD, associate professor at Temple University School of Pharmacy, has received a two-year R21 Research Grant from the National Institutes of Health's National Institute on Aging, to fund a project titled "Comprehensive CRISPR Editing of iPSCs for Dynamic Disease Modeling in Early-Onset Alzheimer's Research."

Early-onset Alzheimer's disease (EOAD) causes cognitive decline before age 65 and can lead to death within 10 to 15 years of symptom onset. No disease-modifying treatments currently exist. One reason progress has been slow is because EOAD damages neurons through several mechanisms simultaneously, including tau abnormalities, impaired autophagy and mitochondrial dysfunction. Until now, researchers haven't had a way to watch all of these processes unfold together, over time, in living cells because most microscopic studies rely on fixed cells and immunostaining, which capture only a single moment rather than an ongoing disease process.

Dr. Perez-Leal's lab will address that gap by building the first comprehensive human live-cell models of EOAD, using a multiplex CRISPR gene-editing technology developed in his lab. The approach introduces a disease-causing mutation into human stem cells while simultaneously tagging several of the cells' own proteins with fluorescent markers. The result is a living cell in which multiple disease pathways can be tracked in real time, without the staining or immunofluorescence techniques that typically limit researchers to a single snapshot. Because the edited cells are otherwise identical to healthy control cells, any changes that emerge can be attributed directly to the disease-causing mutation.

Using this platform, the lab will follow disease development at two critical points: as stem cells differentiate into neurons, when the earliest defects may appear, and in mature neurons, where the disease's more established damage takes hold. Researchers will then use the models to test compounds that target pathways beyond amyloid to determine whether those compounds can prevent early damage, reverse later damage, or both.

"This research will give us a much clearer picture of how early-onset Alzheimer's actually develops, not just what it looks like once damage has already occurred," said Perez-Leal. "By modeling multiple disease pathways simultaneously in living neurons, we hope to identify new therapeutic targets and accelerate the search for treatments that go beyond amyloid."

Completion of the project is expected to advance understanding of EOAD pathophysiology and provide the field with new tools for identifying therapeutic compounds capable of addressing multiple disease pathways at once.

Dr. Perez-Leal's work adds to a growing portfolio of neurodegenerative disease research at Temple University School of Pharmacy, where faculty are pursuing new approaches to some of the most challenging conditions in medicine. The two-year grant will support the project through 2028.