In vitro methods of identifying modulators of neuromuscular junction activity

A neuromuscular junction, activity technology, applied in the field of identification of modulators of neuromuscular and/or muscle activity, can solve the problem that the ability to functionally connect and control the function of human skeletal muscle has not been evaluated or explored.

Active Publication Date: 2018-08-03
MEMORIAL SLOAN KETTERING CANCER CENT
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  • Claims
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Problems solved by technology

However, one of the most important properties of neurons, their ability to form functional synapses and transmit information to suitable downstream targets, remains largely unexplored in human organoids and other PSC-based model systems
[0009] Despite advances in the generation of spinal motor neurons from human PSCs (Amoroso et al., 2013; Calder et al., 2015; Chan et al., 2007; Davis-Dusenbery et al., 2014; Maury et al., 2015; Patani et al., 2011), they The ability to functionally connect and control human skeletal muscle function has not been assessed

Method used

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  • In vitro methods of identifying modulators of neuromuscular junction activity
  • In vitro methods of identifying modulators of neuromuscular junction activity
  • In vitro methods of identifying modulators of neuromuscular junction activity

Examples

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Embodiment 1

[0105] 6.1 Example 1: Modeling of human neuromuscular disease using in vitro neuromuscular junctions under optogenetic control

[0106] Summarize

[0107] Capturing the full potential of human pluripotent stem cell (PSC)-derived neurons in disease modeling and regenerative medicine requires their examination in complex functional systems. Here we demonstrate the establishment of optogenetic control in human PSC-derived spinal motor neurons. Co-cultures of these motor neurons with human myoblast-derived skeletal muscle can be triggered to twitch upon light stimulation. Using Physiological and Imaging Methods to Characterize Newly Established Fully Human Neuromuscular Junctions. To mimic neuromuscular disease, we incubated these co-cultures with IgG and active complement from patients with myasthenia gravis. Myasthenia gravis is an autoimmune disease that selectively targets the neuromuscular junction. We observed a reversible decrease in muscle contraction amplitude, wh...

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Abstract

The present invention relates to an in vitro human neuromuscular junction model prepared from a co-culture of human pluripotent stem cell (PSC)-derived spinal motorneurons and human myoblast-derived skeletal muscle cells. The present invention also provides for methods of screening compounds for their ability to modulate neuromuscular junction activity by determining whether a candidate compound increases or decreases the activity of the in vitro human neuromuscular junction model.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Application No. 62 / 238,531, filed October 7, 2015, which is incorporated by reference in its entirety. [0003] Funding information [0004] This invention was made with government support under Grant No. NS052671 awarded by the National Institutes of Health and the National Institute of Neurological Disorders and Stroke. The government has certain rights in this invention. technical field [0005] The present invention relates to methods for identifying modulators of neuromuscular and / or muscle activity using in vitro models of the human neuromuscular junction. Background technique [0006] Neuromuscular disorders are impairments of motor neuron and / or muscle function due to loss of motor neurons or muscle cells, decreased function of motor neurons or muscle cells, or degenerative changes in central (CNS) or peripheral (PNS) nervous system motor pathways. damagi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0793C12N5/0789
CPCC12N5/0619C12N2501/15C12N2501/155C12N2501/16C12N2501/41C12N2502/1335C12N2506/02C12N5/0607C12N5/0658G01N33/5032G01N33/5058G01N33/5061
Inventor L·施图德J·施泰因贝克
Owner MEMORIAL SLOAN KETTERING CANCER CENT
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