Modeling Hypoxia-Induced Neuropathies Using a Fast and Scalable Human Motor Neuron Differentiation System

• Published in: Stem cell reports 2020-06, Vol.14 (6), p.1033-1043
• Main Authors: Hudish, Laura I., Bubak, Andrew, Triolo, Taylor M., Niemeyer, Christy S., Lorberbaum, David S., Sussel, Lori, Nagel, Maria, Taliaferro, J. Matthew, Russ, Holger A.
• Format: Article
• Language: English
• Subjects: Action Potentials; cell compartments; Cell Hypoxia; Cell Line; Cells, Cultured; diabetic neuropathies; Diabetic Neuropathies - metabolism; fractionation; Humans; hypoxia; Induced Pluripotent Stem Cells - cytology; Mitochondria - metabolism; Mitochondrial Proteins - genetics; Mitochondrial Proteins - metabolism; Motor Neurons - cytology; Motor Neurons - metabolism; Motor Neurons - physiology; neurites; Neuronal Outgrowth; Oxygen - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; soma; stem cell-derived human motor neurons
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2020.04.003

Human motor neuron (MN) diseases encompass a spectrum of disorders. A critical barrier to dissecting disease mechanisms is the lack of appropriate human MN models. Here, we describe a scalable, suspension-based differentiation system to generate functional human MN diseases in 3 weeks. Using this model, we translated recent findings that mRNA mis-localization plays a role in disease development to the human context by establishing a membrane-based system that allows efficient fractionation of MN cell soma and neurites. In response to hypoxia, used to mimic diabetic neuropathies, MNs upregulated mitochondrial transcripts in neurites; however, mitochondria were decreased. These data suggest that hypoxia may disrupt translation of mitochondrial mRNA, potentially leading to neurite damage and development of neuropathies. We report the development of a novel human MN model system to investigate mechanisms of disease affecting soma and/or neurites that facilitates the rapid generation and testing of patient-specific MN diseases. [Display omitted] •Differentiation of human pluripotent stem cells into motor neurons in 3 weeks•Global transcriptomes of motor neuron soma and neurite fractionations•Modeling of diabetic neuropathies by hypoxia reveals neurite-specific defects•Study suggests mRNA mis-localization as novel disease mechanism Russ and colleagues present here a rapid, scalable, suspension-culture system for the efficient generation of functional human motor neurons from pluripotent stem cells. Using a soma and neurite fractionation approach, they reveal that mitochondrial transcripts, but not proteins, are neurite enriched under conditions modeling diabetic neuropathies. These findings suggest mRNA mis-localizations as a potential novel disease mechanism in motor neurons.