A high throughput drug screening assay to identify compounds that promote oligodendrocyte differentiation using acutely dissociated and purified oligodendrocyte precursor cells.
BACKGROUND
Multiple sclerosis is caused by an autoimmune response resulting in demyelination and neural degeneration. The adult central nervous system has the capacity to remyelinate axons in part through the generation of new oligodendrocytes (OLs). To identify clinical candidate compounds that may promote remyelination, we have developed a high throughput screening (HTS) assay to identify compounds that promote the differentiation of oligodendrocyte precursor cells (OPCs) into OLs.


RESULTS
Using acutely dissociated and purified rat OPCs coupled with immunofluorescent image quantification, we have developed an OL differentiation assay. We have validated this assay with a known promoter of differentiation, thyroid hormone, and subsequently used the assay to screen the NIH clinical collection library. We have identified twenty-seven hit compounds which were validated by dose response analysis and the generation of half maximal effective concentration (EC50) values allowed for the ranking of efficacy. The assay identified novel promoters of OL differentiation which we attribute to (1) the incorporation of an OL toxicity pre-screen to allow lowering the concentrations of toxic compounds and (2) the utilization of freshly purified, non-passaged OPCs. These features set our assay apart from other OL differentiation assays used for drug discovery efforts.


CONCLUSIONS
This acute primary OL-based differentiation assay should be of use to those interested in screening large compound libraries for the identification of drugs for the treatment of MS and other demyelinating diseases.
Translational Medicine Center, Myelin Repair Foundation, Sunnyvale, CA, 94085, USA.|Teva Pharmaceuticals, Biologics and CNS Discovery, Redwood City, CA, 94063, USA.
Karen D
KD
Lariosa-Willingham
Translational Medicine Center, Myelin Repair Foundation, Sunnyvale, CA, 94085, USA.|Alios BioPharma, South San Francisco, CA, 94080, USA.
Elen S
ES
Rosler
Translational Medicine Center, Myelin Repair Foundation, Sunnyvale, CA, 94085, USA.
Jay S
JS
Tung
Translational Medicine Center, Myelin Repair Foundation, Sunnyvale, CA, 94085, USA.|Rigel Pharmaceuticals, South San Francisco, CA, 94080, USA.
Jason C
JC
Dugas
Translational Medicine Center, Myelin Repair Foundation, Sunnyvale, CA, 94085, USA.|NGM Biopharmaceuticals, Inc., South San Francisco, CA, 94080, USA.
Tassie L
TL
Collins
Translational Medicine Center, Myelin Repair Foundation, Sunnyvale, CA, 94085, USA. dmitri.leonoudakis@tevapharm.com.|Teva Pharmaceuticals, Biologics and CNS Discovery, Redwood City, CA, 94063, USA. dmitri.leonoudakis@tevapharm.com.
Dmitri
D
Leonoudakis
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D066298:In Vitro Techniques; D009103:Multiple Sclerosis / Q000188:drug therapy; D009836:Oligodendroglia / Q000166:cytology* / Q000187:drug effects*; D051381:Rats; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects*
Differentiation; Drug screening; High throughput; Image analysis; Multiple sclerosis; Myelin basic protein; Myelination; Oligodendrocyte; Primary cell-based assay