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


RESULTS
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
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.
Letzen
Brian S
BS
Department of Biomedical Engineering, The Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.
Liu
Cyndi
C
Thakor
Nitish V
NV
Gearhart
John D
JD
All
Angelo H
AH
Kerr
Candace L
CL
D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D019070:Cell Lineage / Q000235:genetics; D016000:Cluster Analysis; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D009186:Myelin Sheath / Q000235:genetics; D009836:Oligodendroglia / Q000166:cytology* / Q000378:metabolism*; D025341:Principal Component Analysis; D015203:Reproducibility of Results
D015415:Biomarkers; D035683:MicroRNAs
Dev Dyn. 2009 Nov;238(11):2800-12
19806666
J Neurosci. 2003 Jun 15;23(12):5131-40
12832537
J Mol Neurosci. 2009 May;38(1):2-11
18836851
Ann Neurol. 2009 Dec;66(6):843-57
20035504
FASEB J. 2007 Feb;21(2):415-26
17167072
Int J Neurosci. 2010 Apr;120(4):305-13
20374080
Nature. 2005 Jun 16;435(7044):974-8
15944714
Stem Cells Dev. 2007 Dec;16(6):1003-16
18004940
Oncogene. 2010 May 6;29(18):2638-48
20154725
Development. 2001 Jul;128(13):2545-54
11493571
Cell. 2004 Jan 23;116(2):281-97
14744438
Neuron. 2010 Mar 11;65(5):612-26
20223198
Cancer Res. 2007 Feb 1;67(3):976-83
17283129
Pediatr Res. 2002 Feb;51(2):129-35
11809905
Nat Rev Genet. 2004 May;5(5):396-400
15143321
J Neurosci. 2005 May 11;25(19):4694-705
15888645
Med Electron Microsc. 2001 Sep;34(3):160-4
11793190
Brain Res. 2010 Feb 2;1312:120-6
19948152
Glia. 2005 Mar;49(4):480-91
15578660
Glia. 2010 Jan 15;58(2):157-68
19606501
Adv Exp Med Biol. 2007;604:17-46
17695719
Biochem Biophys Res Commun. 2008 Dec 12;377(2):434-440
18930031
J Cell Mol Med. 2005 Apr-Jun;9(2):303-19
15963251
J Neurosci. 2008 Nov 5;28(45):11720-30
18987208
Glia. 2005 Feb;49(3):385-96
15538751
Trends Neurosci. 1996 Mar;19(3):92-6
9054062
Lab Invest. 2001 Mar;81(3):263-81
11310820
Dis Model Mech. 2009 Mar-Apr;2(3-4):178-88
19259393
J Exp Med. 2008 Oct 27;205(11):2473-81
18838545
Biotechniques. 2003 Feb;34(2):374-8
12613259
Nat Rev Neurosci. 2008 Nov;9(11):839-55
18931697
Nature. 2006 Jan 19;439(7074):283-9
16421561
Brain Pathol. 2010 May;20(3):539-50
19775293
Eur J Cancer. 2010 Jun;46(9):1640-9
20219352
Mol Cancer. 2008 Apr 28;7:35
18442408
BMC Genomics. 2010 Feb 18;11:119
20167119
Nat Biotechnol. 2001 Dec;19(12):1129-33
11731781
Science. 1999 Jul 30;285(5428):754-6
10427001
Genes Chromosomes Cancer. 2009 Jul;48(7):569-82
19396866
J Neurosci Res. 2000 Mar 15;59(6):706-11
10700007
Brain Res. 2008 Oct 21;1236:185-93
18710654
Nature. 2005 Jun 9;435(7043):834-8
15944708
J Cell Sci. 2003 Apr 1;116(Pt 7):1279-89
12615970
Proc Natl Acad Sci U S A. 1979 Jan;76(1):514-7
284369
Cell. 2009 Jan 23;136(2):215-33
19167326
Neuron. 2010 Mar 11;65(5):597-611
20223197
Cell. 2005 Jan 14;120(1):15-20
15652477
J Cell Biochem. 2010 Jan 1;109(1):93-102
19885849
Nature. 2004 Sep 16;431(7006):350-5
15372042
J Neurosci. 2005 Nov 2;25(44):10064-73
16267213
J Transl Med. 2009 Mar 23;7:20
19309508
Cell. 2009 Jul 10;138(1):172-85
19596243
Nat Med. 1999 Dec;5(12):1410-2
10581084
Proc Natl Acad Sci U S A. 2000 May 23;97(11):6126-31
10823956
Mol Cell Biol. 2008 Oct;28(20):6426-38
18710938
J Neurosci Res. 2000 Feb 1;59(3):421-9
10679779
J Neurooncol. 2006 Dec;80(3):235-42
16773221
Eur J Neurosci. 2005 Mar;21(6):1469-77
15845075
Nat Rev Genet. 2001 Feb;2(2):120-9
11253051
Nat Med. 2005 Mar;11(3):277-83
15723074