High-content phenotypic screening and triaging strategy to identify small molecules driving oligodendrocyte progenitor cell differentiation.
Multiple Sclerosis is a demyelinating disease of the CNS and the primary cause of neurological disability in young adults. Loss of myelinating oligodendrocytes leads to neuronal dysfunction and death and is an important contributing factor to this disease. Endogenous oligodendrocyte precursor cells (OPCs), which on differentiation are responsible for replacing myelin, are present in the adult CNS. As such, therapeutic agents that can stimulate OPCs to differentiate and remyelinate demyelinated axons under pathologic conditions may improve neuronal function and clinical outcome. We describe the details of an automated, cell-based, morphometric-based, high-content screen that is used to identify small molecules eliciting the differentiation of OPCs after 3 days. Primary screening was performed using rat CG-4 cells maintained in culture conditions that normally support a progenitor cell-like state. From a library of 73,000 diverse small molecules within the Sanofi collection, 342 compounds were identified that increased OPC morphological complexity as an indicator of oligodendrocyte maturation. Subsequent to the primary high-content screen, a suite of cellular assays was established that identified 22 nontoxic compounds that selectively stimulated primary rat OPCs but not C2C12 muscle cell differentiation. This rigorous triaging yielded several chemical series for further expansion and bio- or cheminformatics studies, and their compelling biological activity merits further investigation.
Lead Generation & Candidate Realization, Sanofi Tucson Innovation Center, Tucson, AZ, USA.
Jane V
JV
Peppard
Lead Generation & Candidate Realization, Sanofi R&D, Bridgewater, NJ, USA.
Catherine A
CA
Rugg
Information Services, Genzyme, Framingham, MA, USA.
Matthew A
MA
Smicker
Lead Generation & Candidate Realization, Sanofi Tucson Innovation Center, Tucson, AZ, USA.
Elaine
E
Powers
Lead Generation & Candidate Realization, Sanofi Tucson Innovation Center, Tucson, AZ, USA.
Erica
E
Harnish
Lead Generation & Candidate Realization, Sanofi Tucson Innovation Center, Tucson, AZ, USA.
Joy
J
Prisco
Lead Generation & Candidate Realization, Sanofi Tucson Innovation Center, Tucson, AZ, USA.
Dragan
D
Cirovic
Lead Generation & Candidate Realization, Sanofi Tucson Innovation Center, Tucson, AZ, USA.
Paul S
PS
Wright
Lead Generation & Candidate Realization, Sanofi Tucson Innovation Center, Tucson, AZ, USA.
Paul R
PR
August
MS and Neurology Clinical, Sanofi-Genzyme, Bridgewater, NJ, USA Karen.chandross@sanofi.com.
Karen J
KJ
Chandross
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D055808:Drug Discovery; D004353:Drug Evaluation, Preclinical; D057166:High-Throughput Screening Assays; D006801:Humans; D009103:Multiple Sclerosis; D058953:Neural Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology* / Q000187:drug effects*; D010641:Phenotype; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D054852:Small Molecule Libraries
D054852:Small Molecule Libraries
CNS repair; Multiple Sclerosis (MS); high-content screening (HCS); oligodendrocyte progenitor cell (OPC); phenotypic screening; remyelination