PhenoTherapeutics

Author Of 1 Presentation

Neuroprotection, Regeneration and/or Remyelination Poster Presentation

P0777 - High-content drug screening in human iPSC-derived oligodendrocytes identifies novel pathways to promote oligodendrocyte differentiation (ID 340)

Speakers
Presentation Number
P0777
Presentation Topic
Neuroprotection, Regeneration and/or Remyelination

Abstract

Background

Oligodendrocytes provide vital support to axons via their myelin sheath however these cells are disrupted in multiple sclerosis (MS) causing demyelination; this eventually results in degeneration. Oligodendrocyte progenitor cells (OPCs) are found throughout the central nervous system (CNS), including MS lesions, and are able to differentiate into oligodendrocytes to remyelinate axons. OPCs may therefore be a potential therapeutic target to stimulate remyelination and slow the progression of MS.

Objectives

Our aim is to identify novel compounds which promote oligodendrocyte differentiation from OPCs to mature, myelinating oligodendrocytes.

Methods

Human induced pluripotent stem cells were differentiated into oligodendrocytes (Magnani et al., Methods Mol Biol. 2019) for high-content screening of a drug library of epigenetic regulators at a final concentration of 1μM. Cells were stained for Myelin Basic Protein (MBP) and then imaged on a Molecular Devices ImageXpress; MBP fluorescence was analysed using MetaXpress analysis software. Explants isolated from new-born mouse cerebellum and attached hindbrain were allowed to myelinate normally over 21 days in vitro before being demyelinated with lysophosphatidyl choline (LPC). They were treated with vehicle and compounds and allowed to remyelinate over 14 days post-LPC.

Results

This high-content screening assay was able to identify one compound that increased the number of MBP-positive oligodendrocytes significantly compared to a vehicle (DMSO) control (Z-score = 3.86). Confirmation, in dose-response, determined the EC100 to be 300nM (p=<0.0001, n=3). Oligodendrocyte lineage profiling showed that the compound acted on oligodendrocyte progenitor cells (p=0.02, n=4) and immature oligodendrocytes (p=0.001, n=4) but not neural progenitor cells (p=0.28, n=4). This increased remyelination was also observed in the mammalian ex vivo model (p=0.0118, N=6).

Conclusions

Our drug screening assay was able to identify a compound which promotes oligodendrocyte differentiation via a potentially novel pathway that has not yet been reported in the literature. This compound was found to act on OPCs, and initial target validation was performed using a mammalian ex vivo model and target analogues.

Collapse