On-Chip Assay for Identifying Candidate Microtubule Binding Drugs to Treat ADRDs

ABSTRACT Microtubules (MTs) are structurally and functionally essential components of neurons, providing spatial organization and ensuring intracellular trafficking. MTs are chemically and functionally heterogeneous structures regulated by a complex molecular machinery that can modulate their composition, chemical modification, and propensity for binding with specific MT-associated proteins (MAPs). Dysfunctions of MAPs are directly associated with different forms of Alzheimer’s disease related disorders (ADRDs). This notion led to the development of compounds restoring the correct functioning of MTs and, therefore, counteracting neurodegeneration. However, currently this effort requires the use of methods that are costly, time consuming and inefficient, as it requires the use of cell cultures and animal models. We propose to develop an on-chip functional assay for identifying candidate MT-binding drugs to treat ADRDs. Our on-chip assay will replicate key features of the neuronal MT microenvironment and will use in vitro generated MTs with kinesin based read out system to measure the ability of potential drugs to stabilize and affect MT transport. We will validate the assay using MT associated protein TAU and known MT stabilizing drugs. The Phase I funding will allow us to develop a proof-of-concept on-chip assay that can be further developed in Phase II. The proposed chip will provide a novel AD/ADRD cell-free in vitro model system that could be used in the translation process to perform high- throughput screening for drug discovery and drug efficacy and functionally validate therapeutic target(s).