title: Driving vascular endothelial cell fate of human multipotent Isl1+ heart progenitors with VEGF modified mRNA.
abstract: Distinct families of multipotent heart progenitors play a central role in the generation of diverse cardiac, smooth muscle and endothelial cell lineages during mammalian cardiogenesis. The identification of precise paracrine signals that drive the cell-fate decision of these multipotent progenitors, and the development of novel approaches to deliver these signals in vivo, are critical steps towards unlocking their regenerative therapeutic potential. Herein, we have identified a family of human cardiac endothelial intermediates located in outflow tract of the early human fetal hearts (OFT-ECs), characterized by coexpression of Isl1 and CD144/vWF. By comparing angiocrine factors expressed by the human OFT-ECs and non-cardiac ECs, vascular endothelial growth factor (VEGF)-A was identified as the most abundantly expressed factor, and clonal assays documented its ability to drive endothelial specification of human embryonic stem cell (ESC)-derived Isl1+ progenitors in a VEGF receptor-dependent manner. Human Isl1-ECs (endothelial cells differentiated from hESC-derived ISL1+ progenitors) resemble OFT-ECs in terms of expression of the cardiac endothelial progenitor- and endocardial cell-specific genes, confirming their organ specificity. To determine whether VEGF-A might serve as an in vivo cell-fate switch for human ESC-derived Isl1-ECs, we established a novel approach using chemically modified mRNA as a platform for transient, yet highly efficient expression of paracrine factors in cardiovascular progenitors. Overexpression of VEGF-A promotes not only the endothelial specification but also engraftment, proliferation and survival (reduced apoptosis) of the human Isl1+ progenitors in vivo. The large-scale derivation of cardiac-specific human Isl1-ECs from human pluripotent stem cells, coupled with the ability to drive endothelial specification, engraftment, and survival following transplantation, suggest a novel strategy for vascular regeneration in the heart.
authors.affiliation: 1] Cardiovascular Research Center, Massachusetts General Hospital, Boston, MA 02114, USA [2] Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 01238, USA [3] Stem Cell and Regenerative Medicine Consortium, LKS Faculty of Medicine, University of Hong Kong, Pokfulam, Hong Kong SAR, China.
authors.forename: Kathy O
authors.initials: KO
authors.lastname: Lui
authors.forename: Lior
authors.initials: L
authors.lastname: Zangi
authors.forename: Eduardo A
authors.initials: EA
authors.lastname: Silva
authors.forename: Lei
authors.initials: L
authors.lastname: Bu
authors.forename: Makoto
authors.initials: M
authors.lastname: Sahara
authors.forename: Ronald A
authors.initials: RA
authors.lastname: Li
authors.forename: David J
authors.initials: DJ
authors.lastname: Mooney
authors.forename: Kenneth R
authors.initials: KR
authors.lastname: Chien
mesh_terms: D002454:Cell Differentiation; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D006321:Heart / Q000196:embryology*; D006801:Humans; D060850:LIM-Homeodomain Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D009206:Myocardium / Q000166:cytology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis / Q000235:genetics / Q000378:metabolism*
chemical_list: D060850:LIM-Homeodomain Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D042461:Vascular Endothelial Growth Factor A; C063843:insulin gene enhancer binding protein Isl-1
references.citation: Circulation. 1999 Jan 19;99(2):201-5
references.pmid: 9892583
references.citation: Dev Biol. 2007 Apr 1;304(1):286-96
references.pmid: 17258700
references.citation: Blood. 2004 Nov 1;104(9):2752-60
references.pmid: 15226175
references.citation: Nat Biotechnol. 2011 Feb;29(2):154-7
references.pmid: 21217696
references.citation: Dev Cell. 2007 Aug;13(2):254-67
references.pmid: 17681136
references.citation: J Exp Med. 2005 Jun 6;201(11):1825-35
references.pmid: 15928198
references.citation: Blood. 2007 Mar 1;109(5):1801-9
references.pmid: 17053059
references.citation: Dev Biol. 2009 Dec 15;336(2):137-44
references.pmid: 19835857
references.citation: J Anat. 2012 Apr;220(4):336-49
references.pmid: 22296102
references.citation: Nat Biotechnol. 2013 Oct;31(10):898-907
references.pmid: 24013197
references.citation: Dev Biol. 1993 Oct;159(2):706-19
references.pmid: 8405690
references.citation: J Immunol Methods. 2009 Mar 15;342(1-2):78-81
references.pmid: 19109967
references.citation: Dev Dyn. 2000 Apr;217(4):327-42
references.pmid: 10767078
references.citation: Dev Cell. 2009 Feb;16(2):180-95
references.pmid: 19217421
references.citation: J Atheroscler Thromb. 2005;12(3):138-42
references.pmid: 16020913
references.citation: Genes Dev. 2001 Sep 15;15(18):2470-82
references.pmid: 11562355
references.citation: Nature. 2009 Jul 2;460(7251):113-7
references.pmid: 19571884
references.citation: Proc Natl Acad Sci U S A. 2009 Jan 20;106(3):814-9
references.pmid: 19129488
references.citation: Dev Cell. 2003 Dec;5(6):877-89
references.pmid: 14667410
references.citation: Proc Natl Acad Sci U S A. 2008 Sep 23;105(38):14347-52
references.pmid: 18794520
references.citation: Nat Rev Genet. 2005 Nov;6(11):826-35
references.pmid: 16304598
references.citation: Cell Stem Cell. 2008 May 8;2(5):497-507
references.pmid: 18462699
references.citation: Dev Biol. 2010 Nov 15;347(2):325-36
references.pmid: 20816797
references.citation: PLoS Genet. 2007 Aug;3(8):e140
references.pmid: 17722983
references.citation: Genes Dev. 2005 Nov 1;19(21):2631-42
references.pmid: 16264195
references.citation: Atherosclerosis. 2012 Aug;223(2):284-90
references.pmid: 22727192
references.citation: Circulation. 2000 Aug 29;102(9):940-2
references.pmid: 10961953
references.citation: Dev Dyn. 2000 Dec;219(4):505-13
references.pmid: 11084650
references.citation: Clin Pharmacol Ther. 2002 Jul;72(1):20-32
references.pmid: 12152001
references.citation: Nature. 2005 Feb 10;433(7026):647-53
references.pmid: 15703750
references.citation: Cell. 2006 Dec 15;127(6):1151-65
references.pmid: 17123592
references.citation: Cardiovasc Res. 2007 Feb 1;73(3):481-7
references.pmid: 17134685
references.citation: Nat Med. 2007 Aug;13(8):952-61
references.pmid: 17660828
references.citation: Circ Res. 2012 Nov 9;111(11):e276-80
references.pmid: 23139287
references.citation: Development. 2011 Nov;138(21):4777-87
references.pmid: 21989917
references.citation: Nature. 2010 Mar 25;464(7288):549-53
references.pmid: 20336138
references.citation: Proc Natl Acad Sci U S A. 2001 May 8;98(10):5780-5
references.pmid: 11331753
references.citation: Cell Stem Cell. 2010 Nov 5;7(5):618-30
references.pmid: 20888316
references.citation: Cardiovasc Res. 2010 Dec 1;88(3):502-11
references.pmid: 20631156
references.citation: Nature. 1999 Oct 14;401(6754):670-7
references.pmid: 10537105
references.citation: Cell Stem Cell. 2008 Apr 10;2(4):320-31
references.pmid: 18397752
references.citation: Cell. 2012 Nov 21;151(5):1083-96
references.pmid: 23178125