Abstract
BackgroundNewly proliferated oligodendrocyte precursor cells (OPCs) migrate and surround lesions of patients with multiple sclerosis (MS) and other demyelinating diseases, but fail to differentiate into oligodendrocytes (OLs) and remyelinate remaining viable axons. The abundance of secreted inflammatory factors within and surrounding these lesions likely plays a major inhibitory role, promoting cell death and preventing OL differentiation and axon remyelination. To identify clinical candidate compounds that may protect existing and differentiating OLs in patients, we have developed a high throughput screening (HTS) assay that utilizes purified rat OPCs.ResultsUsing a fluorescent indicator of cell viability coupled with image quantification, we developed an assay to allow the identification of compounds that promote OL viability and differentiation in the presence of the synergistic inflammatory cytokines, tumor necrosis factor α and interferon-γ. We have utilized this assay to screen the NIH clinical collection library and identify compounds that protect OLs and promote OL differentiation in the presence of these inflammatory cytokines.ConclusionThis primary OL-based cytokine protection assay is adaptable for HTS and may be easily modified for profiling of compounds in the presence of other potentially inhibitory molecules found in MS lesions. This assay should be of use to those interested in identifying drugs for the treatment of MS and other demyelinating diseases.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-016-2219-8) contains supplementary material, which is available to authorized users.
Highlights
Proliferated oligodendrocyte precursor cells (OPCs) migrate and surround lesions of patients with multiple sclerosis (MS) and other demyelinating diseases, but fail to differentiate into oligodendrocytes (OLs) and remyelinate remaining viable axons
While it is logical to expect that increasing the number of mature oligodendrocytes will improve the rate and extent of axonal remyelination, it should be considered that the oligodendrocyte differentiation assays described above assess differentiation of OPCs into oligodendrocytes under healthy conditions
We examined the effects of interferon-γ (IFNγ) in the presence tumor necrosis factor α (TNFα), a combination shown to be synergistically deleterious for other cell types in culture [7]
Summary
Proliferated oligodendrocyte precursor cells (OPCs) migrate and surround lesions of patients with multiple sclerosis (MS) and other demyelinating diseases, but fail to differentiate into oligodendrocytes (OLs) and remyelinate remaining viable axons. The presence of TNFα substantially increased the deleterious effects of IFNγ on both differentiation and viability of cultured oligodendrocytes, and did so at cytokine concentrations that were low enough to be relevant to the in vivo disease state Using this information, we developed an assay suitable for rapid high throughput screening, to identify drugs that protect differentiating oligodendrocytes from cytokine toxicity. We developed an assay suitable for rapid high throughput screening, to identify drugs that protect differentiating oligodendrocytes from cytokine toxicity We used this assay to screen the NIH clinical collection (NCC) library (a library of 727 drugs that have been approved for the treatment of various indications), and report here the identification of several FDA-approved drugs that sustain viability and differentiation capacity of oligodendrocytes in the presence of IFNγ and TNFα in vitro. This assay will be useful for screening novel compound libraries and is adaptable to screening compounds in the presence of other factors present in MS lesions
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