Functional Connectivity under Optogenetic Control Allows Modeling of Human Neuromuscular Disease

Abstract

Summary Capturing the full potential of human pluripotent stem cell (PSC)-derived neurons in disease modeling and regenerative medicine requires analysis in complex functional systems. Here we establish optogenetic control in human PSC-derived spinal motorneurons and show that co-culture of these cells with human myoblast-derived skeletal muscle builds a functional all-human neuromuscular junction that can be triggered to twitch upon light stimulation. To model neuromuscular disease we incubated these co-cultures with IgG from myasthenia gravis patients and active complement. Myasthenia gravis is an autoimmune disorder that selectively targets neuromuscular junctions. We saw a reversible reduction in the amplitude of muscle contractions, representing a surrogate marker for the characteristic loss of muscle strength seen in this disease. The ability to recapitulate key aspects of disease pathology and its symptomatic treatment suggests that this neuromuscular junction assay has significant potential for modeling of neuromuscular disease and regeneration.

Autoantibody pathogenicity in a multifocal motor neuropathy induced pluripotent stem cell–derived model

“…Disease modeling using iPSC-derived MNs has previously been applied to studying the underlying disease mechanisms of ALS, and more recently myasthenia gravis. [14][15][16][17][18][19]49 Here we show the potential of iPSC modeling of rare inflammatory disorders. Future improvements to the currently described disease model could be the addition of a coculture with iPSC-derived Schwann cells, thereby providing in vitro myelination and formation of nodes of Ranvier.…”

MicroRNAs Induce a Permissive Chromatin Environment that Enables Neuronal Subtype-Specific Reprogramming of Adult Human Fibroblasts

“…Human myotubes were generated by differentiating human myoblasts using defined culture conditions (Steinbeck et al, 2016). Briefly, human skeletal myoblasts were cultured according to manufacturers recommendations (HSMM; Lonza, CC-2580) then were plated on matrigel (0.1mg/mL) coated 12mm glass coverslips at a density of 80,000 cells/well.…”

Functional Coupling with Cardiac Muscle Promotes Maturation of hPSC-Derived Sympathetic Neurons

“…Moreover, it remains unknown whether a defined population of peripheral neurons can be isolated from hPSCs. Although a recently report presented the generation of functional neuromuscular junction using hPSCs-derived spinal motor neurons and skeletal muscle fibers (Steinbeck et al, 2016), it remains unknown whether hPSC-derived peripheral neurons can mimic neuromodulatory function with their target tissues.…”