title: Genome-Scale Mapping of MicroRNA Signatures in Human Embryonic Stem Cell Neurogenesis.
abstract: To date, lacking of a clinically-suitable source of engraftable human stem/progenitor cells with adequate neurogenic potential has been the major setback in developing effective cell-based therapies against a wide range of neurological disorders. Derivation of human embryonic stem cells (hESCs) provides a powerful tool to investigate the molecular controls in human embryonic neurogenesis as well as an unlimited source to generate the diversity of human neuronal cell types in the developing CNS for repair. However, realizing the developmental and therapeutic potential of hESCs has been hindered by conventional multi-lineage differentiation of pluripotent cells, which is uncontrollable, inefficient, highly variable, difficult to reproduce and scale-up. We recently identified retinoic acid (RA) as sufficient to induce the specification of neuroectoderm direct from the pluripotent state of hESCs under defined platform and trigger progression to human neuronal progenitors (hESC-I hNuPs) and neurons (hESC-I hNus) in the developing CNS with high efficiency, which enables hESC neuronal lineage-specific differentiation and opens the door to investigate human embryonic neurogenesis using the hESC model system. In this study, genome-scale profiling of microRNA (miRNA) differential expression patterns in hESC neuronal lineage-specific progression was used to identify molecular signatures of human embryonic neurogenesis. These in vitro neuroectoderm-derived human neuronal cells have acquired a neuron al identity by down-regulating pluripotence-associated miRNAs and inducing the expression of miRNAs linked to regulating human CNS development to high levels in a stage-specific manner, including silencing of the prominent pluripotence-associated hsa-miR-302 family and drastic expression increases of the Hox hsa-miR-10 and let-7 miRNAs. Following transplantation, hESC-I hNuPs engrafted and yielded well-integrated neurons at a high prevalence within neurogenic regions of the brain. In 3D culture, these hESC-I hNuPs proceeded to express subtype neuronal markers, such as dopaminergic and motor neurons, demonstrating their therapeutic potential for CNS repair. Our study provides critical insight into molecular neurogenesis in human embryonic development as well as offers an adequate human neurogenic cell source in high purity and large quantity for scale-up CNS regeneration.
authors.affiliation: San Diego Regenerative Medicine Institute, San Diego, CA 92109, USA ; Xcelthera, San Diego, CA 92109, USA.
authors.forename: Xuejun H
authors.initials: XH
authors.lastname: Parsons
authors.forename: James F
authors.initials: JF
authors.lastname: Parsons
authors.forename: Dennis A
authors.initials: DA
authors.lastname: Moore
keywords: CNS disease; CNS repair; Cell therapy; Dopaminergic neuron; Embryogenesis; Genome-scale mapping; Human development; Human embryonic stem cell; Let-7; Motor neuron; Neuroectoderm; Neurogenesis; Neuron; Neuronal lineage-specific differentiation; Neuronal progenitor; Pluripotence; miR-10; miR-302; microRNA
references.citation: J Vis Exp. 2011 Oct 28;(56):e3273
references.pmid: 22064669
references.citation: Nat Med. 2006 Nov;12(11):1259-68
references.pmid: 17057709
references.citation: Proc Natl Acad Sci U S A. 2008 Oct 28;105(43):16707-12
references.pmid: 18922775
references.citation: Science. 1998 Nov 6;282(5391):1145-7
references.pmid: 9804556
references.citation: Nature. 2007 Aug 2;448(7153):553-60
references.pmid: 17603471
references.citation: Cell. 2009 May 15;137(4):647-58
references.pmid: 19409607
references.citation: J Vis Exp. 2011 Nov 03;(57):e3274
references.pmid: 22083019
references.citation: Proc Natl Acad Sci U S A. 2010 Feb 2;107(5):1876-81
references.pmid: 20133835
references.citation: Anat Physiol. 2012 Apr 26;(S3):
references.pmid: 23543848
references.citation: Cell Death Differ. 2010 Feb;17(2):209-14
references.pmid: 19461655
references.citation: J Stem Cell Res Ther. 2012 Jul 13;2(3):
references.pmid: 23355957
references.citation: Recent Pat Regen Med. 2011;1(2):142-163
references.pmid: 23355961
references.citation: Cell. 2008 Aug 8;134(3):521-33
references.pmid: 18692474
references.citation: Curr Med Chem. 2011;18(36):5655-65
references.pmid: 22172070
references.citation: Stem Cells. 2004;22(7):1246-55
references.pmid: 15579643
references.citation: Proc Natl Acad Sci U S A. 2007 Jul 17;104(29):12175-80
references.pmid: 17586681
references.citation: Nat Biotechnol. 2011 Aug 10;29(9):824-8
references.pmid: 21832997
references.citation: Stem Cells. 2008 Jun;26(6):1506-16
references.pmid: 18403753
references.citation: Cell. 2006 Apr 21;125(2):315-26
references.pmid: 16630819
references.citation: Cytotherapy. 2009;11(7):815-24
references.pmid: 19903095
references.citation: Cell Stem Cell. 2008 Mar 6;2(3):219-29
references.pmid: 18371447
references.citation: Nat Rev Genet. 2008 Oct;9(10):789-96
references.pmid: 18781158
references.citation: Nat Biotechnol. 2001 Dec;19(12):1129-33
references.pmid: 11731781
references.citation: Nature. 2011 Nov 06;480(7378):547-51
references.pmid: 22056989
references.citation: Nature. 2010 Feb 4;463(7281):621-6
references.pmid: 20054295
references.citation: Dev Cell. 2010 Apr 20;18(4):510-25
references.pmid: 20412767
references.citation: Proc Natl Acad Sci U S A. 2004 Aug 24;101(34):12543-8
references.pmid: 15310843
references.citation: Cell Stem Cell. 2010 Jul 2;7(1):31-5
references.pmid: 20621047
references.citation: Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3225-30
references.pmid: 19218428
references.citation: Proc Natl Acad Sci U S A. 2011 Oct 25;108(43):17803-8
references.pmid: 21969586
references.citation: J Regen Med Tissue Eng. 2012 Sep 23;1(1):
references.pmid: 23542901
references.citation: Curr Opin Cell Biol. 2009 Jun;21(3):461-9
references.pmid: 19278845
references.citation: Nature. 1994 Aug 18;370(6490):567-71
references.pmid: 7914354
references.citation: J Clin Exp Cardiolog. 2012 Apr 25;S9:
references.pmid: 22905333
references.citation: Cell Stem Cell. 2010 Apr 2;6(4):323-35
references.pmid: 20362537
references.citation: Proc Natl Acad Sci U S A. 2010 Jul 20;107(29):13111-6
references.pmid: 20616011
references.citation: Genes Dev. 2008 Jan 15;22(2):152-65
references.pmid: 18198334