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Plate-Based Phenotypic Screening for Pain Using Human iPSC-Derived Sensory Neurons
Peter Stacey1, Anne Mai Wassermann2, Laura Kammonen1
11 Pfizer Neusentis, Great Abington, Cambridgeshire, UK.
This study developed a high-throughput screening method using human induced pluripotent stem cell-derived sensory neurons to identify pain-modulating compounds. The novel assay enables robust phenotypic screening for neuronal excitability, advancing pain research.
Area of Science:
- Neuroscience
- Stem Cell Biology
- Pharmacology
Background:
- Phenotypic screening offers an alternative to target-based drug discovery for complex diseases like pain.
- In vitro phenotypic screens in pain research are limited by the availability of relevant neuronal cell types and functional assays.
Purpose of the Study:
- To establish a scalable, high-throughput phenotypic screening platform for pain research using human induced pluripotent stem cell-derived sensory neurons.
- To develop and validate a functional assay for neuronal excitability relevant to pain and hyperexcitability disorders.
Main Methods:
- Generated human induced pluripotent stem cell (hiPSC)-derived sensory neurons at scale using cryopreserved, near-assay-ready cells.
- Developed a 384-well veratridine-evoked calcium flux assay to measure neuronal excitability.
- Validated the assay's phenotypic relevance to pain and screened 2700 compounds.
Main Results:
- Successfully produced hiPSC sensory neurons in large quantities, decoupling cell production from assay development.
- Established and validated a functional calcium flux assay for neuronal excitability.
- A 2700-compound screen identified diverse target-based mechanisms inhibiting neuronal excitability.
Conclusions:
- High-throughput phenotypic screening using hiPSC sensory neurons is feasible and effective for pain research.
- This platform can identify novel targets, pathways, and compounds for modulating neuronal excitability.
- The developed assay is ready for application in discovering new pain therapeutics.
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