Histone deacetylase activity is required for human oligodendrocyte progenitor differentiation

Abstract

The molecular mechanisms controlling human oligodendrocyte development are poorly characterized. Microarray analysis of human oligodendrocyte progenitor cells (OPCs) and immature oligodendrocytes revealed that specific-class I histone deacetylase (HDAC) target genes were actively repressed during oligodendrocyte commitment. Although epigenetic regulation of oligodendrocyte differentiation has been established in rodent development, the role of HDACs in human OPCs remains undefined. We used HDAC inhibitors (HDACi) trichostatin A (TSA) and sodium butyrate to determine the importance of HDAC activity in human primary OPC differentiation. Treatment with either drug resulted in significant dose-dependent inhibition of O4(+) oligodendrocyte cell differentiation, reduction of oligodendrocyte morphological maturation, and downregulation of myelin basic protein mRNA. High dose TSA treatment was also associated with reduction in OPC proliferation. HDACi treatment prevented downregulation of SOX2, ID4, and TCF7L2 mRNAs but did not regulate HES5, suggesting that targets of HDAC repression may differ between species. These results predict that HDACi treatment would impair proliferation and differentiation by parenchymal oligodendrocyte progenitors, and thereby degrade their potential for endogenous repair in human demyelinating disease. © 2012 Wiley Periodicals, Inc.

Transcription factor induction of human oligodendrocyte progenitor fate and differentiation

“…Immediately after CD133/CD140a FACS, RNA extraction, firststrand synthesis, and qPCR were performed, as described previously (16). Human primers for SYBR Green-based PCR and predesigned primer and Taqman probes were purchased from IDT and Invitrogen, respectively (Table S2).…”

“…To determine the expression profile of infected cells, we performed Illumina microarray analysis (n = 2 fetal samples) and combined these data with the expression profile of CD133/CD140a sorted cells (n = 3 fetal samples) (15). RNA was amplified and Illumina HT-12v4 bead arrays analysis was performed using R/Bioconductor, as described previously (16). We identified genes as significantly regulated using a moderated t test statistic (P < 0.01) and further filtered these genes to examine only those regulated by two or more.…”

“…Given the species differences in OPC gene expression (14), it is likely that the role of these TFs is subtly different in human cells. To select instructive TFs in an unbiased manner, we performed microarray analysis on antigenically defined human progenitors (15,16). We compared the transcriptional profile of CD140a and O4-defined OPCs with that of CD133 + CD140a − NPCs.…”

CD133/CD140a-Based Isolation of Distinct Human Multipotent Neural Progenitor Cells and Oligodendrocyte Progenitor Cells

“…Cells were seeded at 5 · 10 4 /mL into 24-well plates coated with poly-L-ornithine and laminin in SFM lacking growth factors to promote differentiation. OLIG2/O4 immunocytochemistry was performed as described in Conway et al [9], additional staining was performed as described. The proportion of OLIG2 and O4-stained cells was quantified in 10 random fields at 200 · magnification (Olympus IX51), representative of over 200 random cells.…”

“…RNA was amplified and Illumina HT-12v4 bead arrays analysis performed using R/Bioconductor as described in Conway et al [9]. Gene set enrichment analysis (GSEA) was performed [11] using mSigDb [12], Biocarta, GO, and KEGG databases.…”