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MicroRNA expression profiling of oligodendrocyte differentiation from human embryonic stem cells.

Letzen BS, Liu C, Thakor NV, Gearhart JD, All AH, Kerr CL - PLoS ONE (2010)

Bottom Line: A comparison of miRNAs from our cultured OLs and OL progenitors showed significant similarities with published results from equivalent cells found in the rat and mouse central nervous system.These results were supported by correlation analyses between adjacent stages.Our results reveal pronounced trends in miRNA expression and their potential mRNA target interactions that could provide valuable insight into the molecular mechanisms of differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Engineering, The Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.

ABSTRACT

Background: Cells of the oligodendrocyte (OL) lineage play a vital role in the production and maintenance of myelin, a multilamellar membrane which allows for saltatory conduction along axons. These cells may provide immense therapeutic potential for lost sensory and motor function in demyelinating conditions, such as spinal cord injury, multiple sclerosis, and transverse myelitis. However, the molecular mechanisms controlling OL differentiation are largely unknown. MicroRNAs (miRNAs) are considered the "micromanagers" of gene expression with suggestive roles in cellular differentiation and maintenance. Although unique patterns of miRNA expression in various cell lineages have been characterized, this is the first report documenting their expression during oligodendrocyte maturation from human embryonic stem (hES) cells. Here, we performed a global miRNA analysis to reveal and identify characteristic patterns in the multiple stages leading to OL maturation from hES cells including those targeting factors involved in myelin production.

Methodology/principal findings: We isolated cells from 8 stages of OL differentiation. Total RNA was subjected to miRNA profiling and validations preformed using real-time qRT-PCR. A comparison of miRNAs from our cultured OLs and OL progenitors showed significant similarities with published results from equivalent cells found in the rat and mouse central nervous system. Principal component analysis revealed four main clusters of miRNA expression corresponding to early, mid, and late progenitors, and mature OLs. These results were supported by correlation analyses between adjacent stages. Interestingly, the highest differentially-expressed miRNAs demonstrated a similar pattern of expression throughout all stages of differentiation, suggesting that they potentially regulate a common target or set of targets in this process. The predicted targets of these miRNAs include those with known or suspected roles in oligodendrocyte development and myelination including C11Orf9, CLDN11, MYTL1, MBOP, MPZL2, and DDR1.

Conclusions/significance: We demonstrate miRNA profiles during distinct stages in oligodendroglial differentiation that may provide key markers of OL maturation. Our results reveal pronounced trends in miRNA expression and their potential mRNA target interactions that could provide valuable insight into the molecular mechanisms of differentiation.

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Related in: MedlinePlus

Top differentially expressed miRNAs at early neural progenitor stage transitions.Fold change values (log2) and expression profiles of the top ten downregulated miRNAs at the (a) ES-EB and (b) EB-NP stage transitions and the top ten corresponding upregulated miRNAs at the (c) ES-EB and (d) EB-NP stage transitions.
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pone-0010480-g005: Top differentially expressed miRNAs at early neural progenitor stage transitions.Fold change values (log2) and expression profiles of the top ten downregulated miRNAs at the (a) ES-EB and (b) EB-NP stage transitions and the top ten corresponding upregulated miRNAs at the (c) ES-EB and (d) EB-NP stage transitions.

Mentions: Our results show dynamic trends in miRNA expression during OL differentiation strongly suggesting, as it has in other systems, that miRNAs play an important role in regulating differentiation. Specifically, miRNAs which display significant changes at stage-specific transitions are likely to have key roles in regulating this process [10]. Thus, we focused our attention to miRNAs which demonstrated the greatest positive and negative fold changes at each stage transition. Figure 5 summarizes the top ten increasing and decreasing miRNAs at early stage transitions of oligodendrocyte differentiation. At the ESC-EB stage transition, the top 10 miRNAs showing the greatest reduction in expression experienced a ∼3.8-fold change in expression. Similarly, the EB-NP stage transition showed modest downregulation, with a maximum ∼2.8-fold repression. Conversely, the top differentially upregulated miRNAs at these two stage transitions showed much greater changes, with a maximum ∼14-fold change at the ESC-EB transition and a maximum ∼6.8-fold change at the EB-NP transition.


MicroRNA expression profiling of oligodendrocyte differentiation from human embryonic stem cells.

Letzen BS, Liu C, Thakor NV, Gearhart JD, All AH, Kerr CL - PLoS ONE (2010)

Top differentially expressed miRNAs at early neural progenitor stage transitions.Fold change values (log2) and expression profiles of the top ten downregulated miRNAs at the (a) ES-EB and (b) EB-NP stage transitions and the top ten corresponding upregulated miRNAs at the (c) ES-EB and (d) EB-NP stage transitions.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2864763&req=5

pone-0010480-g005: Top differentially expressed miRNAs at early neural progenitor stage transitions.Fold change values (log2) and expression profiles of the top ten downregulated miRNAs at the (a) ES-EB and (b) EB-NP stage transitions and the top ten corresponding upregulated miRNAs at the (c) ES-EB and (d) EB-NP stage transitions.
Mentions: Our results show dynamic trends in miRNA expression during OL differentiation strongly suggesting, as it has in other systems, that miRNAs play an important role in regulating differentiation. Specifically, miRNAs which display significant changes at stage-specific transitions are likely to have key roles in regulating this process [10]. Thus, we focused our attention to miRNAs which demonstrated the greatest positive and negative fold changes at each stage transition. Figure 5 summarizes the top ten increasing and decreasing miRNAs at early stage transitions of oligodendrocyte differentiation. At the ESC-EB stage transition, the top 10 miRNAs showing the greatest reduction in expression experienced a ∼3.8-fold change in expression. Similarly, the EB-NP stage transition showed modest downregulation, with a maximum ∼2.8-fold repression. Conversely, the top differentially upregulated miRNAs at these two stage transitions showed much greater changes, with a maximum ∼14-fold change at the ESC-EB transition and a maximum ∼6.8-fold change at the EB-NP transition.

Bottom Line: A comparison of miRNAs from our cultured OLs and OL progenitors showed significant similarities with published results from equivalent cells found in the rat and mouse central nervous system.These results were supported by correlation analyses between adjacent stages.Our results reveal pronounced trends in miRNA expression and their potential mRNA target interactions that could provide valuable insight into the molecular mechanisms of differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Engineering, The Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.

ABSTRACT

Background: Cells of the oligodendrocyte (OL) lineage play a vital role in the production and maintenance of myelin, a multilamellar membrane which allows for saltatory conduction along axons. These cells may provide immense therapeutic potential for lost sensory and motor function in demyelinating conditions, such as spinal cord injury, multiple sclerosis, and transverse myelitis. However, the molecular mechanisms controlling OL differentiation are largely unknown. MicroRNAs (miRNAs) are considered the "micromanagers" of gene expression with suggestive roles in cellular differentiation and maintenance. Although unique patterns of miRNA expression in various cell lineages have been characterized, this is the first report documenting their expression during oligodendrocyte maturation from human embryonic stem (hES) cells. Here, we performed a global miRNA analysis to reveal and identify characteristic patterns in the multiple stages leading to OL maturation from hES cells including those targeting factors involved in myelin production.

Methodology/principal findings: We isolated cells from 8 stages of OL differentiation. Total RNA was subjected to miRNA profiling and validations preformed using real-time qRT-PCR. A comparison of miRNAs from our cultured OLs and OL progenitors showed significant similarities with published results from equivalent cells found in the rat and mouse central nervous system. Principal component analysis revealed four main clusters of miRNA expression corresponding to early, mid, and late progenitors, and mature OLs. These results were supported by correlation analyses between adjacent stages. Interestingly, the highest differentially-expressed miRNAs demonstrated a similar pattern of expression throughout all stages of differentiation, suggesting that they potentially regulate a common target or set of targets in this process. The predicted targets of these miRNAs include those with known or suspected roles in oligodendrocyte development and myelination including C11Orf9, CLDN11, MYTL1, MBOP, MPZL2, and DDR1.

Conclusions/significance: We demonstrate miRNA profiles during distinct stages in oligodendroglial differentiation that may provide key markers of OL maturation. Our results reveal pronounced trends in miRNA expression and their potential mRNA target interactions that could provide valuable insight into the molecular mechanisms of differentiation.

Show MeSH
Related in: MedlinePlus