Transcription factor rational design improves directed differentiation of human mesenchymal stem cells into skeletal myocytes.
There is great interest in transdifferentiating cells from one lineage into those of another and in dedifferentiating mature cells back into a stem/progenitor cell state by deploying naturally occurring transcription factors (TFs). Often, however, steering cellular differentiation pathways in a predictable and efficient manner remains challenging. Here, we investigated the principle of combining domains from different lineage-specific TFs to improve directed cellular differentiation. As proof-of-concept, we engineered the whole-human TF MyoDCD, which has the NH(2)-terminal transcription activation domain (TAD) and adjacent DNA-binding motif of MyoD COOH-terminally fused to the TAD of myocardin (MyoCD). We found via reporter gene and marker protein assays as well as by a cell fusion readout system that, targeting the TAD of MyoCD to genes normally responsive to the skeletal muscle-specific TF MyoD enforces more robust myogenic reprogramming of nonmuscle cells than that achieved by the parental, prototypic master TF, MyoD. Human mesenchymal stem cells (hMSCs) transduced with a codon-optimized microdystrophin gene linked to a synthetic striated muscle-specific promoter and/or with MyoD or MyoDCD were evaluated for complementing the genetic defect in Duchenne muscular dystrophy (DMD) myocytes through heterotypic cell fusion. Cotransduction of hMSCs with MyoDCD and microdystrophin led to chimeric myotubes containing the highest dystrophin levels.
Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands. m.goncalves@lumc.nl
Manuel A F V
MA
Gonçalves
Josephine M
JM
Janssen
Quynh G
QG
Nguyen
Takis
T
Athanasopoulos
Stephen D
SD
Hauschka
George
G
Dickson
Antoine A F
AA
de Vries
D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D005434:Flow Cytometry; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D018485:Muscle Fibers, Skeletal / Q000166:cytology* / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
D014157:Transcription Factors
Nature. 1990 Mar 22;344(6264):348-51
2179732
Exp Cell Res. 2008 May 1;314(8):1721-33
18395202
Proc Natl Acad Sci U S A. 2006 Nov 7;103(45):16995-7000
17077152
Nature. 2009 Dec 3;462(7273):587-94
19956253
FASEB J. 2007 Oct;21(12):3369-79
17579192
Hum Mol Genet. 2008 Aug 15;17(16):2507-17
18511457
Hum Mol Genet. 2010 Mar 1;19(5):752-60
19965907
Blood. 2004 Jul 1;104(1):290-4
15010375
Curr Biol. 2003 Jan 21;13(2):105-15
12546783
Neuromuscul Disord. 2007 Apr;17(4):276-84
17303423
J Biol Chem. 2008 Dec 12;283(50):34720-7
18945675
Mol Ther. 2008 Apr;16(4):657-64
18334986
Mol Ther. 2008 Apr;16(4):741-8
18334989
PLoS One. 2010 Aug 18;5(8):e12239
20805873
Mol Cell Biol. 2004 Sep;24(18):8264-75
15340086
Nat Med. 2008 Feb;14(2):134-43
18204461
Mol Ther. 2008 Nov;16(11):1825-32
18766174
Mol Ther. 2007 Feb;15(2):320-9
17235310
Mol Ther. 2008 Dec;16(12):1944-52
18827804
Mol Ther. 2009 Jun;17(6):1064-72
19352326
Brain Dev. 2006 Aug;28(7):420-5
16584859
Hum Mol Genet. 2006 Jan 15;15(2):213-21
16321987
Science. 1999 Apr 2;284(5411):143-7
10102814
Mol Cell Biol. 2005 Jan;25(1):364-76
15601857
Somat Cell Mol Genet. 1990 Nov;16(6):557-65
2267630
Expert Opin Biol Ther. 2009 Jul;9(7):849-66
19527108
Neuromuscul Disord. 2002 Oct;12 Suppl 1:S3-6
12206788
Mol Ther. 2007 May;15(5):867-77
17387336
Nature. 2001 Jun 28;411(6841):1014-5
11429592
N Engl J Med. 1993 Sep 23;329(13):915-20
8361505
J Clin Invest. 1998 May 15;101(10):2119-28
9593768
Lancet. 1999 Feb 27;353(9154):727-8
10073525
J Biol Chem. 2005 Aug 5;280(31):28749-60
15834131
Neuromuscul Disord. 1996 May;6(3):195-202
8784808
PLoS One. 2010 Jun 03;5(6):e10954
20532169
J Cell Mol Med. 2009 Nov-Dec;13(11-12):4385-402
19602034
PLoS One. 2007 Aug 22;2(8):e774
17712422
Nature. 2008 Oct 2;455(7213):627-32
18754011
Neuromuscul Disord. 2010 Jan;20(1):6-15
20034794
Hum Gene Ther. 2003 Nov 1;14(16):1525-33
14577914
Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):4065-70
17360478
J Neurochem. 2006 Feb;96(3):777-89
16405511
Nature. 2010 Feb 25;463(7284):1035-41
20107439
Mol Ther. 2007 Feb;15(2):431-8
17235323