High-Throughput In Vitro Analyses of Trauma-Induced Tauopathy

PROJECT SUMMARY Chronic traumatic encephalopathy (CTE) is a neurodegenerative disease associated with repeated mild traumatic brain injury (TBI). CTE is among the many neurodegenerative diseases characterized as tauopathies, wherein the protein tau, which is usually associated with microtubules in the axons of neurons, becomes separated from microtubules, initiating a degenerative cascade and leading to eventual neurofunctional loss. There are currently no pharmacological treatments available for CTE patients, so any treatment that could limit or reverse tau-associated dysfunction would have an important impact on TBI patient outcomes. Moreover, given the similarity between CTE and other tauopathies, insights into CTE treatment could be broadly applicable to other common neurodegenerative diseases. We have recently developed an in vitro model that directly links mechanical injury to tau pathology in cultured neurons. One notable outcome from our prior studies is that we found that synaptic dysfunction (measured using patch clamp) is correlated with mislocalization of tau to dendritic spines (measured using fluorescent imaging). This result suggests that this relatively simple image-based readout could be used for screening the effects of pharmacological agents on the functional progression of trauma-induced tauopathy. There are currently no high-throughput in vitro models for screening the effects of pharmaceuticals on trauma-induced tauopathy. We have developed several cell stretching systems, both for neurotrauma and other applications. However, all of these systems are far too low-throughput for performing drug discovery studies. Thus, our goal is to scale up our current in vitro neurotrauma model to a high-enough throughput for drug screening studies. We will design a new stretchable multi-well plate for neuronal cell culture and a new high strain rate stretcher that can apply trauma-like loads to the cells in the plate. In addition, we will employ machine learning based algorithms to quickly and efficiently analyze the data collected from our new device. Finally, we will use the device to test inhibitors known to be effective against other aspects of neuronal injury affect tau mislocalization