Phase separation in Alzheimer's disease and Phase-separated c-Abl dynamics

Neurodegenerative diseases like Alzheimer's disease (AD) are a major global health concern, affecting millions and placing significant economic burdens on healthcare systems. Despite advancements in understanding AD pathogenesis, effective treatments remain elusive. PhaSeAD (Phase separation in Alzheimer's disease and Phase-separated c-Abl dynamics) aims to explore the role of the c-Abl protein kinase in AD pathogenesis, focusing on its interaction with liquid-liquid phase separation (LLPS). Objectives include understanding c-Abl phosphorylation specificity using NMR spectroscopy to study its interaction with substrates Tau and PSD-95, providing insights into c-Abl's substrate specificity and phosphorylation selectivity. Additionally, we will explore c-Abl function within LLPS droplets using biochemical assays like ITC and kinase assays to assess its activity inside and outside LLPS droplets. Fluorescence microscopy techniques like FRAP and FCS will visualize and quantify c-Abl dynamics within LLPS droplets and its interaction with other LLPS components. Furthermore, we will investigate the role of c-Abl and LLPS in regulating cytoskeletal dynamics, crucial for neuronal function and disrupted in AD. High-resolution microscopy techniques, including super-resolution and electron microscopy, will visualize and quantify the impact of c-Abl and LLPS on cytoskeletal assembly and organization. Live-cell imaging techniques will assess their impact on neuronal morphology and function. Expected outcomes include a comprehensive understanding of c-Abl's behavior within LLPS droplets, its impact on Tau and PSD-95 phosphorylation, and its regulatory role in cytoskeletal homeostasis. These findings will enhance our understanding of c-Abl's involvement in AD pathogenesis and may lead to novel therapeutic targets for developing effective treatments for AD and other neurodegenerative diseases.