High-throughput screening in embryonic stem cell-derived neurons identifies potentiators of alpha-amino-3-hydroxyl-5-methyl-4-isoxazolepropionate-type glutamate receptors.
Stem cell biology offers advantages to investigators seeking to identify new therapeutic molecules. Specifically, stem cells are genetically stable, scalable for molecular screening, and function in cellular assays for drug efficacy and safety. A key hurdle for drug discoverers of central nervous system disease is a lack of high quality neuronal cells. In the central nervous system, alpha-amino-3-hydroxyl-5-methyl-4-isoxazolepropionate (AMPA) subtype glutamate receptors mediate the vast majority of excitatory neurotransmissions. Embryonic stem (ES) cell protocols were developed to differentiate into neuronal subtypes that express AMPA receptors and were pharmacologically responsive to standard compounds for AMPA potentiation. Therefore, we hypothesized that stem cell-derived neurons should be predictive in high-throughput screens (HTSs). Here, we describe a murine ES cell-based HTS of a 2.4 x 10(6) compound library, the identification of novel chemical "hits" for AMPA potentiation, structure function relationship of compounds and receptors, and validation of chemical leads in secondary assays using human ES cell-derived neurons. This reporting of murine ES cell derivatives being formatted to deliver HTS of greater than 10(6) compounds for a specific drug target conclusively demonstrates a new application for stem cells in drug discovery. In the future new molecular entities may be screened directly in human ES or induced pluripotent stem cell derivatives.
McNeish
John
J
FrGenetically Modified Models COE, Pfizer Global Research and Development, Groton, Connecticut 06340, USA. john.d.mcneish@pfizer.com
Roach
Marsha
M
Hambor
John
J
Mather
Robert J
RJ
Weibley
Laura
L
Lazzaro
John
J
Gazard
Justin
J
Schwarz
Jacob
J
Volkmann
Robert
R
Machacek
David
D
Stice
Steve
S
Zawadzke
Laura
L
O'Donnell
Christopher
C
Hurst
Raymond
R
D000818:Animals; D002626:Chemistry, Pharmaceutical / Q000379:methods; D015195:Drug Design; D004353:Drug Evaluation, Preclinical; D053595:Embryonic Stem Cells / Q000166:cytology*; D005470:Fluorometry / Q000379:methods; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D008954:Models, Biological; D009154:Mutation; D009474:Neurons / Q000378:metabolism*; D018091:Receptors, AMPA / Q000737:chemistry*; D017470:Receptors, Glutamate / Q000378:metabolism*; D013678:Technology, Pharmaceutical / Q000379:methods; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid / Q000494:pharmacology*
D018091:Receptors, AMPA; D017470:Receptors, Glutamate; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid