Human motor units in microfluidic devices are impaired by < /i>FUS< //i> mutations and improved by HDAC6 inhibition

Dittlau, Katarina Stoklund; Krasnow, Emily N; Fumagalli, Laura; Vandoorne, Tijs; Baatsen, Pieter; Kerstens, Axelle; Giacomazzi, Giorgia; Pavie, Benjamin; Rossaert, Elisabeth; Beckers, Jimmy; Sampaolesi, Maurilio; Van Damme, Philip; Van den Bosch, Ludo

doi:10.1016/j.stemcr.2021.03.029

Human motor units in microfluidic devices are impaired by < /i>FUS< //i> mutations and improved by HDAC6 inhibition

Author:

Dittlau, Katarina Stoklund

Krasnow, Emily N ; Fumagalli, Laura ; Vandoorne, Tijs ; Baatsen, Pieter ; Kerstens, Axelle ; Giacomazzi, Giorgia ; Pavie, Benjamin ; Rossaert, Elisabeth ; Beckers, Jimmy ; Sampaolesi, Maurilio ; Van Damme, Philip ; Van den Bosch, Ludo

Keywords:

Science & Technology, Life Sciences & Biomedicine, Cell & Tissue Engineering, Cell Biology, NEUROMUSCULAR-JUNCTION FORMATION, AMYOTROPHIC-LATERAL-SCLEROSIS, HUMAN SKELETAL-MUSCLE, MODEL, ABNORMALITIES, MOTONEURONS, LAMININ, SYSTEM, AGRIN, FUS, HDAC6, Tubastatin A, amyotrophic lateral sclerosis, microfluidic device, neurite outgrowth, neurite regrowth, neuromuscular junction, Agrin, Amyotrophic Lateral Sclerosis, Biomarkers, Cell Culture Techniques, Cell Differentiation, Coculture Techniques, Fluorescent Antibody Technique, Histone Deacetylase 6, Histone Deacetylase Inhibitors, Humans, Induced Pluripotent Stem Cells, Lab-On-A-Chip Devices, Laminin, Microfluidic Analytical Techniques, Motor Neurons, Muscle Fibers, Skeletal, Mutation, Neuromuscular Junction, Neuronal Outgrowth, RNA-Binding Protein FUS, C14/17/107#54271215, C14/17/111#54271220, 0601 Biochemistry and Cell Biology, 1103 Clinical Sciences, 3101 Biochemistry and cell biology

Abstract:

Neuromuscular junctions (NMJs) ensure communication between motor neurons (MNs) and muscle; however, in MN disorders, such as amyotrophic lateral sclerosis (ALS), NMJs degenerate resulting in muscle atrophy. The aim of this study was to establish a versatile and reproducible in vitro model of a human motor unit to investigate the effects of ALS-causing mutations. Therefore, we generated a co-culture of human induced pluripotent stem cell (iPSC)-derived MNs and human primary mesoangioblast-derived myotubes in microfluidic devices. A chemotactic and volumetric gradient facilitated the growth of MN neurites through microgrooves resulting in the interaction with myotubes and the formation of NMJs. We observed that ALS-causing FUS mutations resulted in reduced neurite outgrowth as well as an impaired neurite regrowth upon axotomy. NMJ numbers were likewise reduced in the FUS-ALS model. Interestingly, the selective HDAC6 inhibitor, Tubastatin A, improved the neurite outgrowth, regrowth, and NMJ morphology, prompting HDAC6 inhibition as a potential therapeutic strategy for ALS.