A multilevel screening pipeline in zebrafish identifies therapeutic drugs for GAN.

Giant axonal neuropathy (GAN) is a fatal neurodegenerative disorder for which there is currently no treatment. Affecting the nervous system, GAN starts in infancy with motor deficits that rapidly evolve toward total loss of ambulation. Using the gan zebrafish model that reproduces the loss of motility as seen in patients, we conducted the first pharmacological screening for the GAN pathology. Here, we established a multilevel pipeline to identify small molecules restoring both the physiological and the cellular deficits in GAN. We combined behavioral, in silico, and high‐content imaging analyses to refine our Hits to five drugs restoring locomotion, axonal outgrowth, and stabilizing neuromuscular junctions in the gan zebrafish. The postsynaptic nature of the drug's cellular targets provides direct evidence for the pivotal role the neuromuscular junction holds in the restoration of motility. Our results identify the first drug candidates that can now be integrated in a repositioning approach to fasten therapy for the GAN disease. Moreover, we anticipate both our methodological development and the identified hits to be of benefit to other neuromuscular diseases. Synopsis: To date, there is no treatment for giant axonal neuropathy (GAN), a fatal neurodegenerative disease leading to loss of ambulation in young patients. Due to the lack of effective rodent models, the gan zebrafish was used to conduct the first pharmacological screening for therapeutic repurposing.The gan zebrafish is the first robust model of the GAN disease, exhibiting the loss of motility found in patients.A multi‐level methodology was designed to integrate behavioral (motility), computational (drug‐target functions) and cellular (NMJ compartments) readouts.A novel high‐content automated imaging analysis was developed to quantify the beneficial effect of the Hits at the NMJ.This tailored screening pipeline identified five Hits restoring the motility of the gan zebrafish through the stabilization of the NMJ. [ABSTRACT FROM AUTHOR]