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Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Thomas, Douglas D. / Turk, Boris / Wittinghofer, Alfred

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Volume 397, Issue 5


Biomechanical and biochemical regulation of cathepsin K expression in endothelial cells converge at AP-1 and NF-κB

Philip M. Keegan
  • The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology/Emory University, 950 Atlantic Drive, Suite 3015, Atlanta, GA 30332, USA
  • Other articles by this author:
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/ Suhaas Anbazhakan
  • The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology/Emory University, 950 Atlantic Drive, Suite 3015, Atlanta, GA 30332, USA
  • Other articles by this author:
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/ Baolin Kang / Betty S. Pace / Manu O. Platt
  • Corresponding author
  • The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology/Emory University, 950 Atlantic Drive, Suite 3015, Atlanta, GA 30332, USA
  • Email
  • Other articles by this author:
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Published Online: 2016-01-12 | DOI: https://doi.org/10.1515/hsz-2015-0244


Cathepsins K and V are powerful elastases elevated in endothelial cells by tumor necrosis factor-α (TNFα) stimulation and disturbed blood flow both of which contribute to inflammation-mediated arterial remodeling. However, mechanisms behind endothelial cell integration of biochemical and biomechanical cues to regulate cathepsin production are not known. To distinguish these mechanisms, human aortic endothelial cells (HAECs) were stimulated with TNFα and exposed to pro-remodeling or vasoprotective shear stress profiles. TNFα upregulated cathepsin K via JNK/c-jun activation, but vasoprotective shear stress inhibited TNFα-stimulated cathepsin K expression. JNK/c-jun were still phosphorylated, but cathepsin K mRNA levels were significantly reduced to almost null indicating separate biomechanical regulation of cathepsin K by shear stress separate from biochemical stimulation. Treatment with Bay 11-7082, an inhibitor of IκBα phosphorylation, was sufficient to block induction of cathepsin K by both pro-remodeling shear stress and TNFα, implicating NF-κB as the biomechanical regulator, and its protein levels were reduced in HAECs by vasoprotective shear stress. In conclusion, NF-κB and AP-1 activation were necessary to activate cathepsin K expression in endothelial cells, highlighting integration of biochemical and biomechanical stimuli to control cathepsins K and V, powerful elastases implicated for arterial remodeling due to chronic inflammation and disturbed blood flow.

Keywords: proteases; shear stress; sickle cell disease; TNF-α


  • Barabino, G.A., Platt, M.O., and Kaul, D.K. (2010). Sickle cell biomechanics. Annu. Rev. Biomed. Eng. 12, 345–367.Web of ScienceCrossrefGoogle Scholar

  • Belcher, J.D., Mahaseth, H., Welch, T.E., Vilback, A.E., Sonbol, K.M., Kalambur, V.S., Bowlin, P.R., Bischof, J.C., Hebbel, R.P., and Vercellotti, G.M. (2005). Critical role of endothelial cell activation in hypoxia-induced vasoocclusion in transgenic sickle mice. Am. J. Physiol. Heart Circ. Physiol. 288, H2715–H2725.Google Scholar

  • Boros, L., Thomas, C., and Weiner, W.J. (1976). Large cerebral vessel disease in sickle cell anaemia. J. Neurol. Neurosurg. Psychiatry 39, 1236–1239.CrossrefGoogle Scholar

  • Castier, Y., Ramkhelawon, B., Riou, S., Tedgui, A., and Lehoux, S. (2009). Role of NF-κB in flow-induced vascular remodeling. Antioxid Redox Signal 11, 1641–1649.Google Scholar

  • Chaar, V., Picot, J., Renaud, O., Bartolucci, P., Nzouakou, R., Bachir, D., Galacteros, F., Colin, Y., Le Van Kim, C., and El Nemer, W. (2010). Aggregation of mononuclear and red blood cells through an α4β1-Lu/basal cell adhesion molecule interaction in sickle cell disease. Haematologica 95, 1841–1848.Web of ScienceGoogle Scholar

  • Chapman, H.A., Riese, R.J., and Shi, G.P. (1997). Emerging roles for cysteine proteases in human biology. Annu. Rev. Physiol. 59, 63–88.Google Scholar

  • Cuhlmann, S., Van der Heiden, K., Saliba, D., Tremoleda, J.L., Khalil, M., Zakkar, M., Chaudhury, H., Luong le, A., Mason, J.C., Udalova, I., et al. (2011). Disturbed blood flow induces RelA expression via c-Jun N-terminal kinase 1: a novel mode of NF-κB regulation that promotes arterial inflammation. Circ. Res. 108, 950–959.Google Scholar

  • Dai, G., Kaazempur-Mofrad, M.R., Natarajan, S., Zhang, Y., Vaughn, S., Blackman, B.R., Kamm, R.D., Garcia-Cardena, G., and Gimbrone, M.A., Jr. (2004). Distinct endothelial phenotypes evoked by arterial waveforms derived from atherosclerosis-susceptible and -resistant regions of human vasculature. Proc. Natl. Acad. Sci. USA 101, 14871–14876.CrossrefGoogle Scholar

  • Frenette, P.S. (2004). Sickle cell vasoocclusion: heterotypic, multicellular aggregations driven by leukocyte adhesion. Microcirculation 11, 167–177.CrossrefGoogle Scholar

  • Fujioka, S., Niu, J., Schmidt, C., Sclabas, G.M., Peng, B., Uwagawa, T., Li, Z., Evans, D.B., Abbruzzese, J.L., and Chiao, P.J. (2004). NF-κB and AP-1 connection: mechanism of NF-κB-dependent regulation of AP-1 activity. Mol. Cell Biol. 24, 7806–7819.CrossrefGoogle Scholar

  • Gareus, R., Kotsaki, E., Xanthoulea, S., van der Made, I., Gijbels, M.J., Kardakaris, R., Polykratis, A., Kollias, G., de Winther, M.P., and Pasparakis, M. (2008). Endothelial cell-specific NF-κB inhibition protects mice from atherosclerosis. Cell Metab. 8, 372–383.CrossrefGoogle Scholar

  • Gelb, B.D., Shi, G.P., Heller, M., Weremowicz, S., Morton, C., Desnick, R.J., and Chapman, H.A. (1997). Structure and chromosomal assignment of the human cathepsin K gene. Genomics 41, 258–262.CrossrefGoogle Scholar

  • Glagov, S., Rowley, D.A., and Kohut, R.I. (1961). Atherosclerosis of human aorta and its coronary and renal arteries. A consideration of some hemodynamic factors which may be related to the marked differences in atherosclerotic involvement of the coronary and renal arteries. Arch. Pathol. 72, 558–571.Google Scholar

  • Hajra, L., Evans, A.I., Chen, M., Hyduk, S.J., Collins, T., and Cybulsky, M.I. (2000). The NF-κB signal transduction pathway in aortic endothelial cells is primed for activation in regions predisposed to atherosclerotic lesion formation. Proc. Natl. Acad. Sci. USA 97, 9052–9057.CrossrefGoogle Scholar

  • Hassler, O. (1962). The windows of the internal elastic lamella of the cerebral arteries. Virchow’s Arch. Pathol. Anat. Physiol. Klin. Med. 335, 127–132.Google Scholar

  • Hillery, C. and Panepinto, J. (2004). Pathophysiology of stroke in sickle cell disease. Microcirculation 11, 195–208.CrossrefGoogle Scholar

  • Keegan, P.M., Surapaneni, S., and Platt, M.O. (2012a). Sickle cell disease activates peripheral blood mononuclear cells to induce cathepsins k and v activity in endothelial cells. Anemia 2012, 201781.Google Scholar

  • Keegan, P.M., Wilder, C.L., and Platt, M.O. (2012b). Tumor necrosis factor alpha stimulates cathepsin K and V activity via juxtacrine monocyte-endothelial cell signaling and JNK activation. Mol. Cell Biochem. 367, 65–72.Web of ScienceGoogle Scholar

  • Ku, D.N. (1997). Blood flow in arteries. Annu. Rev. Fluid Mech. 29, 399–434.Web of ScienceGoogle Scholar

  • Ku, D.N. and Giddens, D.P. (1987). Laser Doppler anemometer measurements of pulsatile flow in a model carotid bifurcation. J. Biomech. 20, 407–421.CrossrefGoogle Scholar

  • Li, Z., Yasuda, Y., Li, W., Bogyo, M., Katz, N., Gordon, R.E., Fields, G.B., and Bromme, D. (2004). Regulation of collagenase activities of human cathepsins by glycosaminoglycans. J. Biol. Chem. 279, 5470–5479.Google Scholar

  • Li, W.A., Barry, Z.T., Cohen, J.D., Wilder, C.L., Deeds, R.J., Keegan, P.M., and Platt, M.O. (2010). Detection of femtomole quantities of mature cathepsin K with zymography. Anal. Biochem. 401, 91–98.Web of ScienceGoogle Scholar

  • Lutgens, E., Lutgens, S.P., Faber, B.C., Heeneman, S., Gijbels, M.M., de Winther, M.P., Frederik, P., van der Made, I., Daugherty, A., Sijbers, A.M., et al. (2006). Disruption of the cathepsin K gene reduces atherosclerosis progression and induces plaque fibrosis but accelerates macrophage foam cell formation. Circulation 113, 98–107.CrossrefGoogle Scholar

  • Malavé, I., Perdomo, Y., Escalona, E., Rodriguez, E., Anchustegui, M., Malavé, H., and Arends, T. (1993). Levels of tumor necrosis factor α/cachectin (TNFα) in sera from patients with sickle cell disease. Acta Haematol. 90, 172–176.Google Scholar

  • Merkel, K.H., Ginsberg, P.L., Parker, J.C., and Post, M.J. (1978). Cerebrovascular disease in sickle cell anemia: a clinical, pathological and radiological correlation. Stroke 9, 45–52.CrossrefGoogle Scholar

  • Pang, M., Martinez, A.F., Fernandez, I., Balkan, W., and Troen, B.R. (2007). AP-1 stimulates the cathepsin K promoter in RAW 264.7 cells. Gene 403, 151–158.Web of ScienceGoogle Scholar

  • Platt, O.S. (2005). Preventing stroke in sickle cell anemia. N. Engl. J. Med. 353, 2743–2745.Google Scholar

  • Platt, M.O., Ankeny, R.F., and Jo, H. (2006). Laminar shear stress inhibits cathepsin L activity in endothelial cells. Arterioscler. Thromb. Vasc. Biol. 26, 1784–1790.CrossrefGoogle Scholar

  • Platt, M.O., Ankeny, R.F., Shi, G.P., Weiss, D., Vega, J.D., Taylor, W.R., and Jo, H. (2007). Expression of cathepsin K is regulated by shear stress in cultured endothelial cells and is increased in endothelium in human atherosclerosis. Am. J. Physiol. Heart Circ. Physiol. 292, H1479–H1486.Google Scholar

  • Sorescu, G.P., Song, H., Tressel, S.L., Hwang, J., Dikalov, S., Smith, D.A., Boyd, N.L., Platt, M.O., Lassegue, B., Griendling, K.K., et al. (2004). Bone morphogenic protein 4 produced in endothelial cells by oscillatory shear stress induces monocyte adhesion by stimulating reactive oxygen species production from a nox1-based NADPH oxidase. Circ. Res. 95, 773–779.Google Scholar

  • Stein, B., Baldwin, A.S., Jr., Ballard, D.W., Greene, W.C., Angel, P., and Herrlich, P. (1993). Cross-coupling of the NF-κB p65 and Fos/Jun transcription factors produces potentiated biological function. EMBO J 12, 3879–3891.Google Scholar

  • Stockman, J.A., Nigro, M.A., Mishkin, M.M., and Oski, F.A. (1972). Occlusion of large cerebral vessels in sickle-cell anemia. N. Engl. J. Med. 287, 846–849.Google Scholar

  • Sukhova G.K., Shi, G.P, Simon D.I., Chapman H.A., and Libby, P. (1998). Expression of the elastolytic cathepsins S and K in human atheroma and regulation of their production in smooth muscle cells. J. Clin. Invest. 102, 576–583.CrossrefGoogle Scholar

  • Suo, J., Ferrara, D.E., Sorescu, D., Guldberg, R.E., Taylor, W.R., and Giddens, D.P. (2007). Hemodynamic shear stresses in mouse aortas: implications for atherogenesis. Arterioscler. Thromb. Vasc. Biol. 27, 346–351.Web of ScienceCrossrefGoogle Scholar

  • Switzer, J.A., Hess, D.C., Nichols, F.T., and Adams, R.J. (2006). Pathophysiology and treatment of stroke in sickle-cell disease: present and future. Lancet Neurol. 5, 501–512.CrossrefGoogle Scholar

  • Verma, I.M., Stevenson, J.K., Schwarz, E.M., Van Antwerp, D., and Miyamoto, S. (1995). Rel/NF-κB/IκB family: intimate tales of association and dissociation. Gene. Dev. 9, 2723–2735.CrossrefGoogle Scholar

  • Wang, J., An, F.S., Zhang, W., Gong, L., Wei, S.J., Qin, W.D., Wang, X.P., Zhao, Y.X., Zhang, Y., Zhang, C., et al. (2011). Inhibition of c-Jun N-terminal kinase attenuates low shear stress-induced atherogenesis in apolipoprotein E-deficient mice. Mol. Med. 17, 990–999.Web of ScienceGoogle Scholar

  • Wang, C., Baker, B.M., Chen, C.S., and Schwartz, M.A. (2013). Endothelial cell sensing of flow direction. Arterioscler. Thromb. Vasc. Biol. 33, 2130–2136.Web of ScienceGoogle Scholar

  • Wilder, C.L., Park, K.Y., Keegan, P.M., and Platt, M.O. (2011). Manipulating substrate and pH in zymography protocols selectively distinguishes cathepsins K, L, S, and V activity in cells and tissues. Arch. Biochem. Biophys. 516, 52–57.Web of ScienceGoogle Scholar

  • Zarins, C.K., Giddens, D.P., Bharadvaj, B.K., Sottiurai, V.S., Mabon, R.F., and Glagov, S. (1983). Carotid bifurcation atherosclerosis. Quantitative correlation of plaque localization with flow velocity profiles and wall shear stress. Circ. Res. 53, 502–514.CrossrefGoogle Scholar

About the article

Received: 2015-09-08

Accepted: 2016-01-04

Published Online: 2016-01-12

Published in Print: 2016-05-01

Funding Source: National Institutes of Health

Award identifier / Grant number: 1DP2OD007433-01

This work was supported by the National Institutes of Health [1DP2OD007433-01 to M.O.P] and also by a National Science Foundation Graduate Research Fellowship to P.M.K.

Conflict of Interest statement: None declared.

Citation Information: Biological Chemistry, Volume 397, Issue 5, Pages 459–468, ISSN (Online) 1437-4315, ISSN (Print) 1431-6730, DOI: https://doi.org/10.1515/hsz-2015-0244.

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