Expression of Lineage Transcription Factors Identifies Differences in Transition States of Induced Human Oligodendrocyte Differentiation.

Sabrina Galinski,Michael J Ziller,Andrea Schmitt,Peter Falkai,Laura V Glaser,Nirmal Kannaiyan,Jan Benedikt Waldeck,Marius Stephan,Michael C Wehr,Damianos Demetriou,Florian J Raabe,Moritz J Rossner,Verena Huber

doi:10.3390/cells11020241

Abstract

Oligodendrocytes (OLs) are critical for myelination and are implicated in several brain disorders. Directed differentiation of human-induced OLs (iOLs) from pluripotent stem cells can be achieved by forced expression of different combinations of the transcription factors SOX10 (S), OLIG2 (O), and NKX6.2 (N). Here, we applied quantitative image analysis and single-cell transcriptomics to compare different transcription factor (TF) combinations for their efficacy towards robust OL lineage conversion. Compared with S alone, the combination of SON increases the number of iOLs and generates iOLs with a more complex morphology and higher expression levels of myelin-marker genes. RNA velocity analysis of individual cells reveals that S generates a population of oligodendrocyte-precursor cells (OPCs) that appear to be more immature than those generated by SON and to display distinct molecular properties. Our work highlights that TFs for generating iOPCs or iOLs should be chosen depending on the intended application or research question, and that SON might be beneficial to study more mature iOLs while S might be better suited to investigate iOPC biology.

Highlights

To investigate transcription factor (TF)-based differences of directed differentiation of induced oligodendrocyte precursor cells (iOPCs) and induced oligodendrocytes (iOLs) with S [13], SO [12] and SON [11], we applied these TF combinations in accordance with the neural induction and differentiation protocol adapted from Garcia-Leon et al [13,19]
We confirmed that S, SO- and SON-directed conversion are all sufficient to rapidly generate oligodendroglial lineage cells from human-induced pluripotent stem cells (hiPSCs)
As a result of the side-by-side analysis at the single-cell resolution, we showed that the application of SON generated more O4+ late-stage iOPCs and more mature and complex

Summary

Introduction

Myelinating oligodendrocytes (OLs) are essential for saltatory nerve conduction in the central nervous system and are involved in the metabolic support of neurons and modulation of neuronal excitability [1,2]. OLs are derived from oligodendrocyte precursor cells (OPCs) and both cell types are considered heterogeneous populations with regional specifications and functionally different states [3,4,5,6]. OL dysfunction is associated with several major neurological diseases, e.g., multiple sclerosis, leukodystrophy, stroke, and schizophrenia [1,7]. The advent of human-induced pluripotent stem cells (hiPSCs) paved the way for generating hiPSC-derived OPCs (hiPSC OPCs) and hiPSC-derived OLs (hiPSC OLs) that improved our understanding of human biology and enabled dissection of pathophysiological mechanisms, including cell-based therapies [8,9]

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