Author:

Keywords:

Science & Technology, Life Sciences & Biomedicine, Neurosciences, Neurosciences & Neurology, Amyotrophic lateral sclerosis, Astrocyte, Reactivity, Cytokines, Motor unit, Microfluidic, Neuromuscular junction, WNT/beta-catenin pathway, AMYOTROPHIC-LATERAL-SCLEROSIS, NF-KAPPA-B, SPINAL-CORD, SIGNALING PATHWAY, WNT, EXPRESSION, ACTIVATION, GLUTAMATE, NEURONS, DEGENERATION, WNT/β-catenin pathway, Humans, Amyotrophic Lateral Sclerosis, Astrocytes, Induced Pluripotent Stem Cells, Motor Neurons, Neuromuscular Junction, RNA-Binding Protein FUS, C24E/20/067#55722626, 1133520N|1133522N#54908522, G0D3620N#55524479, 0604 Genetics, 1103 Clinical Sciences, 1109 Neurosciences, Neurology & Neurosurgery, 3101 Biochemistry and cell biology, 3209 Neurosciences

Abstract:

BACKGROUND: Astrocytes play a crucial, yet not fully elucidated role in the selective motor neuron pathology in amyotrophic lateral sclerosis (ALS). Among other responsibilities, astrocytes provide important neuronal homeostatic support, however this function is highly compromised in ALS. The establishment of fully human coculture systems can be used to further study the underlying mechanisms of the dysfunctional intercellular interplay, and has the potential to provide a platform for revealing novel therapeutic entry points. METHODS: In this study, we characterised human induced pluripotent stem cell (hiPSC)-derived astrocytes from FUS-ALS patients, and incorporated these cells into a human motor unit microfluidics model to investigate the astrocytic effect on hiPSC-derived motor neuron network and functional neuromuscular junctions (NMJs) using immunocytochemistry and live-cell recordings. FUS-ALS cocultures were systematically compared to their CRISPR-Cas9 gene-edited isogenic control systems. RESULTS: We observed a dysregulation of astrocyte homeostasis, which resulted in a FUS-ALS-mediated increase in reactivity and secretion of inflammatory cytokines. Upon coculture with motor neurons and myotubes, we detected a cytotoxic effect on motor neuron-neurite outgrowth, NMJ formation and functionality, which was improved or fully rescued by isogenic control astrocytes. We demonstrate that ALS astrocytes have both a gain-of-toxicity and loss-of-support function involving the WNT/β-catenin pathway, ultimately contributing to the disruption of motor neuron homeostasis, intercellular networks and NMJs. CONCLUSIONS: Our findings shine light on a complex, yet highly important role of astrocytes in ALS, and provides further insight in to their pathological mechanisms.