title: c-Myc is essential to prevent endothelial pro-inflammatory senescent phenotype.
abstract: The proto-oncogene c-Myc is vital for vascular development and promotes tumor angiogenesis, but the mechanisms by which it controls blood vessel growth remain unclear. In the present work we investigated the effects of c-Myc knockdown in endothelial cell functions essential for angiogenesis to define its role in the vasculature. We provide the first evidence that reduction in c-Myc expression in endothelial cells leads to a pro-inflammatory senescent phenotype, features typically observed during vascular aging and pathologies associated with endothelial dysfunction. c-Myc knockdown in human umbilical vein endothelial cells using lentivirus expressing specific anti-c-Myc shRNA reduced proliferation and tube formation. These functional defects were associated with morphological changes, increase in senescence-associated-β-galactosidase activity, upregulation of cell cycle inhibitors and accumulation of c-Myc-deficient cells in G1-phase, indicating that c-Myc knockdown in endothelial cells induces senescence. Gene expression analysis of c-Myc-deficient endothelial cells showed that senescent phenotype was accompanied by significant upregulation of growth factors, adhesion molecules, extracellular-matrix components and remodeling proteins, and a cluster of pro-inflammatory mediators, which include Angptl4, Cxcl12, Mdk, Tgfb2 and Tnfsf15. At the peak of expression of these cytokines, transcription factors known to be involved in growth control (E2f1, Id1 and Myb) were downregulated, while those involved in inflammatory responses (RelB, Stat1, Stat2 and Stat4) were upregulated. Our results demonstrate a novel role for c-Myc in the prevention of vascular pro-inflammatory phenotype, supporting an important physiological function as a central regulator of inflammation and endothelial dysfunction.
authors.affiliation: Interdisciplinary Stem Cell Institute, University of Miami Leonard M. Miller School of Medicine, Miami, Florida, United States of America.
authors.forename: Victoria
authors.initials: V
authors.lastname: Florea
authors.forename: Nithya
authors.initials: N
authors.lastname: Bhagavatula
authors.forename: Gordana
authors.initials: G
authors.lastname: Simovic
authors.forename: Francisco Y
authors.initials: FY
authors.lastname: Macedo
authors.forename: Ricardo A
authors.initials: RA
authors.lastname: Fock
authors.forename: Claudia O
authors.initials: CO
authors.lastname: Rodrigues
mesh_terms: D049109:Cell Proliferation; D016922:Cellular Senescence / Q000235:genetics*; D042783:Endothelial Cells / Q000378:metabolism* / Q000473:pathology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D053263:Gene Regulatory Networks; D016259:Genes, myc; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007249:Inflammation / Q000235:genetics* / Q000473:pathology*; D018836:Inflammation Mediators / Q000378:metabolism; D010641:Phenotype; D000090063:Proto-Oncogene Mas; D013312:Stress, Physiological / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D018836:Inflammation Mediators; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D014157:Transcription Factors
references.citation: Am J Pathol. 2006 Jan;168(1):9-19
references.pmid: 16400005
references.citation: Mucosal Immunol. 2011 Mar;4(2):172-85
references.pmid: 20980995
references.citation: Mol Biol Cell. 2007 Feb;18(2):536-46
references.pmid: 17151361
references.citation: PLoS Biol. 2008 Dec 2;6(12):2853-68
references.pmid: 19053174
references.citation: Cell. 2012 Sep 28;151(1):68-79
references.pmid: 23021216
references.citation: Proc Natl Acad Sci U S A. 1987 Aug;84(16):5788-92
references.pmid: 2886992
references.citation: Circulation. 2000 Oct 24;102(17):2111-7
references.pmid: 11044429
references.citation: Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3260-5
references.pmid: 10737792
references.citation: Cytokine Growth Factor Rev. 2011 Jun;22(3):131-9
references.pmid: 21757394
references.citation: Arterioscler Thromb Vasc Biol. 2006 Aug;26(8):1712-20
references.pmid: 16675726
references.citation: Annu Rev Pathol. 2010;5:99-118
references.pmid: 20078217
references.citation: J Biol Chem. 1999 Sep 10;274(37):26141-8
references.pmid: 10473565
references.citation: J Biol Chem. 2011 Oct 21;286(42):36841-51
references.pmid: 21832056
references.citation: J Appl Physiol (1985). 2009 Jan;106(1):326-32
references.pmid: 19036896
references.citation: Proc Natl Acad Sci U S A. 2006 Mar 7;103(10):3645-50
references.pmid: 16537449
references.citation: Gastroenterology. 2008 Aug;135(2):552-67
references.pmid: 18598698
references.citation: Circ Res. 2006 Nov 24;99(11):1207-15
references.pmid: 17068295
references.citation: J Gerontol A Biol Sci Med Sci. 2010 Nov;65(11):1165-80
references.pmid: 20675618
references.citation: Cell Cycle. 2006 Aug;5(15):1621-8
references.pmid: 16921263
references.citation: J Immunol. 2010 Apr 1;184(7):3311-9
references.pmid: 20304832
references.citation: Aging Cell. 2009 Dec;8(6):676-89
references.pmid: 19732043
references.citation: Nat Cell Biol. 2001 Apr;3(4):392-9
references.pmid: 11283613
references.citation: Development. 2008 Aug;135(14):2467-77
references.pmid: 18550710
references.citation: J Exp Med. 2008 May 12;205(5):1037-48
references.pmid: 18411341
references.citation: Development. 2008 Jun;135(11):1903-11
references.pmid: 18469221
references.citation: J Exp Med. 2008 May 12;205(5):1049-62
references.pmid: 18411337
references.citation: Arterioscler Thromb Vasc Biol. 2006 Jul;26(7):1503-9
references.pmid: 16690877
references.citation: Acta Physiol (Oxf). 2009 Jun;196(2):193-222
references.pmid: 19220204
references.citation: Am J Pathol. 1991 Mar;138(3):765-75
references.pmid: 2000945
references.citation: J Exp Med. 2008 Oct 27;205(11):2457-64
references.pmid: 18824582
references.citation: Neoplasia. 2002 May-Jun;4(3):185-90
references.pmid: 11988837
references.citation: Circ Res. 2005 Apr 15;96(7):784-91
references.pmid: 15761195
references.citation: Mol Cancer Res. 2012 Jun;10(6):677-88
references.pmid: 22661548
references.citation: Crit Rev Oncol Hematol. 2008 Apr;66(1):1-9
references.pmid: 17913510
references.citation: Mucosal Immunol. 2011 Mar;4(2):186-96
references.pmid: 20962771
references.citation: J Immunol. 2003 Nov 1;171(9):4868-74
references.pmid: 14568967
references.citation: Biochem Biophys Res Commun. 2010 Jan 22;391(4):1737-41
references.pmid: 20043872
references.citation: Nat Med. 2011 Jan;17(1):117-22
references.pmid: 21151138
references.citation: Nat Cell Biol. 2010 Jan;12(1):54-9; sup pp 1-14
references.pmid: 20010815
references.citation: Eur J Cancer. 2013 Jan;49(1):219-30
references.pmid: 22683307
references.citation: J Vasc Res. 2010;47(1):80-90
references.pmid: 19729955
references.citation: Clin Hemorheol Microcirc. 2002;26(2):117-23
references.pmid: 12082260
references.citation: Proc Natl Acad Sci U S A. 2000 Aug 15;97(17):9498-503
references.pmid: 10920185
references.citation: Cell Signal. 2011 Jul;23(7):1076-81
references.pmid: 21144894
references.citation: Cytokine. 2005 Mar 7;29(5):229-35
references.pmid: 15760679
references.citation: Nat Genet. 2000 Feb;24(2):171-4
references.pmid: 10655064
references.citation: Am J Physiol Heart Circ Physiol. 2012 Aug 15;303(4):H429-38
references.pmid: 22707563
references.citation: Genes Dev. 2002 Oct 1;16(19):2530-43
references.pmid: 12368264
references.citation: Antioxid Redox Signal. 2008 Sep;10(9):1631-74
references.pmid: 18598143
references.citation: Nature. 2008 Jul 24;454(7203):436-44
references.pmid: 18650914
references.citation: Pediatr Int. 2010 Feb;52(1):75-9
references.pmid: 19460128
references.citation: ScientificWorldJournal. 2005 Sep 29;5:828-33
references.pmid: 16200329
references.citation: J Vasc Res. 2010;47(4):287-98
references.pmid: 20016203
references.citation: Am J Pathol. 2003 May;162(5):1521-8
references.pmid: 12707035
references.citation: J Vasc Surg. 1993 Aug;18(2):170-6; discussion 176-7
references.pmid: 8350425
references.citation: Circ Res. 2001 Dec 7;89(12):1161-7
references.pmid: 11739281
references.citation: ScientificWorldJournal. 2011;11:2491-505
references.pmid: 22235180
references.citation: Cancer Microenviron. 2012 Dec;5(3):237-47
references.pmid: 22833050
references.citation: Adv Clin Chem. 2010;52:109-30
references.pmid: 21275341
references.citation: Nature. 1996 Aug 8;382(6591):511-7
references.pmid: 8700224
references.citation: Sci Transl Med. 2012 Feb 1;4(119):119ra14
references.pmid: 22301555
references.citation: J Biol Chem. 2007 Nov 30;282(48):34938-44
references.pmid: 17913706
references.citation: Curr Opin Cell Biol. 2010 Oct;22(5):617-25
references.pmid: 20817428
references.citation: Circulation. 2001 Nov 27;104(22):2673-8
references.pmid: 11723017
references.citation: Arch Biochem Biophys. 2003 Aug 15;416(2):129-36
references.pmid: 12893289
references.citation: Circ Res. 2009 May 22;104(10):1151-9
references.pmid: 19407242
references.citation: Pathol Int. 2012 Jul;62(7):445-55
references.pmid: 22726064
references.citation: Curr Oncol Rep. 2011 Feb;13(1):71-6
references.pmid: 21080117
references.citation: Nature. 2001 May 17;411(6835):342-8
references.pmid: 11357141
references.citation: Microcirculation. 2003 Jun;10(3-4):359-70
references.pmid: 12851652
references.citation: Trends Mol Med. 2010 May;16(5):238-46
references.pmid: 20444648
references.citation: FASEB J. 1999 Jan;13(1):181-9
references.pmid: 9872942
references.citation: Neoplasia. 2008 Apr;10(4):340-7
references.pmid: 18392135
references.citation: J Am Soc Nephrol. 2008 May;19(5):953-60
references.pmid: 18287561
references.citation: J Vasc Surg. 2006 Sep;44(3):633-41
references.pmid: 16950446
references.citation: Curr Opin Endocrinol Diabetes Obes. 2013 Apr;20(2):81-6
references.pmid: 23422245
references.citation: J Surg Res. 1996 Mar;61(2):323-9
references.pmid: 8656603
references.citation: Cell Cycle. 2010 Sep 15;9(18):3655-61
references.pmid: 20818171
references.citation: Nat Med. 2003 Jun;9(6):653-60
references.pmid: 12778163