Danio rerio: a “humanized” in vivo model for the study of non-cell autonomous mechanisms in ALS

Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disorder characterized by the loss of both upper and lower motor neurons (MNs). In this regard, in the last years the non-cell autonomous contribution of reactive astrocytes has been considered as extremely influential to the course of the pathology. Unfortunately, in-vitro 2D cultures and mouse models are not capable to fully recapitulate molecular pathological features and motor deficits observed in patients, which instead unfolds in ALS zebrafish models. Furthermore, a remarkable similarity with human genome and its central nervous system architecture, along with a large spawning number and the ease of being subjected to transplantations, allow us to consider zebrafish as a reliable in vivo model to study ALS. At the state of art, the project aims to understand the role of non-cell autonomous mechanisms by injecting mutant iPSC-derived astrocytes and isogenic controls into the spinal cord of WT zebrafish strains, thus assessing the molecular and phenotypical aberrations sufficiently induced by reactive astrocyte. Particularly, the analysis will be focused on evaluating the presence of neuronal functional and structural impairment, muscular atrophy, motor behaviour induced by mutant astrocytes transplanted. To obtain a more comprehensive investigation, high-throughput spatial transcriptomic analysis will be performed, thus giving a deep insight into non-cell autonomous pathological mechanisms in environmental and spatial conditions similar to humans one. This model will hopefully yield novel insight about the significance contribution of reactive astrocytes to MNs degeneration, thus as a screening hub for effective therapeutics that can shorten the path from preclinical to clinical research phase of ALS