Heparan sulfate proteoglycans mediate interstitial flow mechanotransduction regulating MMP-13 expression and cell motility via FAK-ERK in 3D collagen.
BACKGROUND
Interstitial flow directly affects cells that reside in tissues and regulates tissue physiology and pathology by modulating important cellular processes including proliferation, differentiation, and migration. However, the structures that cells utilize to sense interstitial flow in a 3-dimensional (3D) environment have not yet been elucidated. Previously, we have shown that interstitial flow upregulates matrix metalloproteinase (MMP) expression in rat vascular smooth muscle cells (SMCs) and fibroblasts/myofibroblasts via activation of an ERK1/2-c-Jun pathway, which in turn promotes cell migration in collagen. Herein, we focused on uncovering the flow-induced mechanotransduction mechanism in 3D.


RESULTS
Cleavage of rat vascular SMC surface glycocalyx heparan sulfate (HS) chains from proteoglycan (PG) core proteins by heparinase or disruption of HS biosynthesis by silencing N-deacetylase/N-sulfotransferase 1 (NDST1) suppressed interstitial flow-induced ERK1/2 activation, interstitial collagenase (MMP-13) expression, and SMC motility in 3D collagen. Inhibition or knockdown of focal adhesion kinase (FAK) also attenuated or blocked flow-induced ERK1/2 activation, MMP-13 expression, and cell motility. Interstitial flow induced FAK phosphorylation at Tyr925, and this activation was blocked when heparan sulfate proteoglycans (HSPGs) were disrupted. These data suggest that HSPGs mediate interstitial flow-induced mechanotransduction through FAK-ERK. In addition, we show that integrins are crucial for mechanotransduction through HSPGs as they mediate cell spreading and maintain cytoskeletal rigidity.


CONCLUSIONS
We propose a conceptual mechanotransduction model wherein cell surface glycocalyx HSPGs, in the presence of integrin-mediated cell-matrix adhesions and cytoskeleton organization, sense interstitial flow and activate the FAK-ERK signaling axis, leading to upregulation of MMP expression and cell motility in 3D. This is the first study to describe a flow-induced mechanotransduction mechanism via HSPG-mediated FAK activation in 3D. This study will be of interest in understanding the flow-related mechanobiology in vascular lesion formation, tissue morphogenesis, cancer cell metastasis, and stem cell differentiation in 3D, and also has implications in tissue engineering.
Department of Biomedical Engineering, The City College of New York, The City University of New York, New York, New York, United States of America.
Zhong-Dong
ZD
Shi
Hui
H
Wang
John M
JM
Tarbell
D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002465:Cell Movement; D003094:Collagen; D003599:Cytoskeleton / Q000378:metabolism; D045604:Extracellular Fluid / Q000502:physiology*; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism*; D019812:Heparan Sulfate Proteoglycans / Q000502:physiology*; D016023:Integrins / Q000378:metabolism; D053509:Matrix Metalloproteinase 13 / Q000235:genetics*; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D008954:Models, Biological; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D051381:Rats; D015854:Up-Regulation / Q000235:genetics
D019812:Heparan Sulfate Proteoglycans; D016023:Integrins; D003094:Collagen; D051416:Focal Adhesion Protein-Tyrosine Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D053509:Matrix Metalloproteinase 13; C507083:Mmp13 protein, rat
J Biol Chem. 2002 Apr 5;277(14):12270-4
11805099
Trends Cell Biol. 2007 Jan;17(1):44-50
17141502
PLoS One. 2009 Aug 10;4(8):e6565
19668337
J Biol Chem. 2007 May 18;282(20):14845-52
17395594
Circulation. 2009 Nov 10;120(19):1883-1892
19858418
Science. 1993 May 21;260(5111):1124-7
7684161
Am J Physiol Heart Circ Physiol. 2009 Oct;297(4):H1196-7
19525374
Nat Clin Pract Cardiovasc Med. 2009 Jan;6(1):16-26
19029993
J Biol Chem. 1989 Feb 25;264(6):3514-23
2521637
J Cell Sci. 2007 Jun 1;120(Pt 11):1829-32
17515480
Exp Cell Res. 2008 Nov 1;314(18):3369-81
18657535
Cell Tissue Res. 2010 Jan;339(1):31-46
19597846
Arterioscler Thromb Vasc Biol. 2009 Apr;29(4):562-70
19150878
Virchows Arch A Pathol Anat Histol. 1981;392(2):199-216
7281508
J Cell Physiol. 2005 Apr;203(1):166-76
15389626
Tissue Eng. 2005 Sep-Oct;11(9-10):1476-88
16259602
Ann Biomed Eng. 2005 Dec;33(12):1719-23
16389519
Science. 2001 Nov 23;294(5547):1708-12
11721053
Nat Rev Mol Cell Biol. 2005 Jan;6(1):56-68
15688067
Am J Physiol Heart Circ Physiol. 2007 Jun;292(6):H3128-35
17308005
PLoS One. 2010 Aug 16;5(8):e12196
20808940
Nature. 1994 Dec 22-29;372(6508):786-91
7997267
Proc Natl Acad Sci U S A. 2009 Dec 29;106(52):22102-7
20080785
J Clin Invest. 2001 Apr;107(8):935-41
11306594
J Biol Chem. 2002 Sep 6;277(36):32970-7
12087088
Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):2805-10
10077592
Arterioscler Thromb Vasc Biol. 2000 Oct;20(10):2220-5
11031207
Annu Rev Biochem. 1999;68:729-77
10872465
Sci Signal. 2008 Oct 07;1(40):pt8
18840877
Am J Physiol Heart Circ Physiol. 2007 Aug;293(2):H1023-30
17468337
Q J Exp Physiol. 1987 Oct;72(4):409-37
3321140
J Cell Biol. 1995 Dec;131(6 Pt 2):1903-15
8557756
Nat Rev Mol Cell Biol. 2009 Jan;10(1):53-62
19197332
Anat Rec. 2000 Jan 1;258(1):1-14
10603443
Trends Cell Biol. 1998 May;8(5):189-92
9695837
Biorheology. 1994 Mar-Apr;31(2):163-70
8729478
J Cell Sci. 2001 Oct;114(Pt 20):3583-90
11707510
Scand J Med Sci Sports. 2009 Aug;19(4):479-89
19538537
Am J Physiol Heart Circ Physiol. 2009 Oct;297(4):H1225-34
19465549
Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3546-51
11891289
Matrix Biol. 2003 Mar;22(1):25-33
12714039
Curr Opin Cell Biol. 2006 Oct;18(5):472-81
16930976
J Appl Physiol (1985). 2005 Jan;98(1):242-9
15322072
Circ Res. 2003 Nov 14;93(10):e136-42
14563712
Cancer Cell. 2007 Jun;11(6):526-38
17560334
Nature. 2007 Apr 26;446(7139):1030-7
17460664
Biochim Biophys Acta. 2009 Oct;1790(10):1314-26
19632304
Proc Natl Acad Sci U S A. 2007 Oct 2;104(40):15941-6
17895377
Int Rev Cytol. 2001;207:113-50
11352265
J Biol Chem. 1997 Nov 28;272(48):30455-62
9374537
Circ J. 2009 Nov;73(11):1983-92
19801852
Annu Rev Biomed Eng. 2007;9:229-56
17459001
Nat Rev Mol Cell Biol. 2007 Dec;8(12):957-69
17971838
Am J Physiol Heart Circ Physiol. 2010 Jan;298(1):H127-35
19880665
Tissue Eng Part A. 2009 Jan;15(1):175-85
18636940
Am J Physiol Cell Physiol. 2008 Jun;294(6):C1350-61
18400991
Curr Opin Genet Dev. 2004 Feb;14(1):92-101
15108811
Genes Cells. 2007 Nov;12(11):1245-54
17986008
Annu Rev Biomed Eng. 2007;9:121-67
17373886
Cell. 1992 Apr 3;69(1):11-25
1555235
Proc Natl Acad Sci U S A. 2008 Mar 11;105(10):3927-32
18310319
Int Immunol. 2008 Mar;20(3):353-63
18182379
J Cell Physiol. 2007 Dec;213(3):565-73
17680633
J Intern Med. 2006 Apr;259(4):339-50
16594902
J Biol Chem. 2004 Jul 16;279(29):30588-99
15096502
Oncogene. 2000 Nov 20;19(49):5606-13
11114741
Biophys J. 2002 Sep;83(3):1650-60
12202388
Biophys J. 2006 Jul 1;91(1):113-21
16603487