Project information
Intracellular Transport Defects in Astrocytes and Neurons Derived from Adar Mutant Mice (ITDANAdarM)

Efficient intracellular transport is fundamental for neuronal and glial function, supporting axonal cargo delivery, organelle distribution, synaptic maintenance, and astrocyte–neuron communication. Disruption of transport pathways is a common early feature of neurodevelopmental and neurodegenerative diseases. Astrocytes, in particular, rely on highly regulated vesicle trafficking for metabolic support and gliotransmission, yet their contribution to ADAR-related pathology remains largely unexplored. This project is based on the central hypothesis that loss of ADAR function leads to cell-autonomous defects in intracellular transport in both neurons and astrocytes, contributing to brain dysfunction independently of inflammatory signaling.
We will investigate this hypothesis using primary neuronal and astrocytic cultures derived from Adar mutant mouse models. We will investigate axonal transport in primary neurons using live-cell imaging of mitochondria and lysosomes in microfluidic chambers, measuring transport parameters such as velocity, run length and directionality. These functional readouts will be correlated with expression changes in motor complex proteins identified in Adar mutant brains. Motor proteins, kinesins and dyneins, are essential also for transporting building blocks of intermediate filament (IF) cytoskeleton along microtubules, thus affecting the overall shape of such IF networks. IF networks build of glial fibrillary acidic protein (GFAP) and vimentin in astrocytes have been shown to impact trafficking of various cytoplasmic vesicles. Other types of IF networks in astrocytes, particularly those expressed in reactive astrocytes, might also be affected by altered expression of molecular motors. Thus, we will next assess cytoplasmic distribution of IF networks and vesicular trafficking in primary astrocytes by tracking endolysosomal vesicle dynamics, providing a detailed characterization of astrocyte-specific transport defects caused by ADAR loss. By combining genetically defined mouse models with high-resolution live imaging, this project will establish intracellular transport as a key functional pathway downstream of ADAR-mediated RNA editing. The expected outcomes will redefine ADAR-related brain pathology by identifying transport impairment as an early and mechanistically distinct contributor to disease, complementing immune-driven mechan.