Original articles
Jingshu Tang, Yuying Kang, Longjian Huang, Lei Wu, Ying Penga. TIMP1 preserves the blood-brain barrier through interacting with CD63/integrin β1 complex and regulating downstream FAK/RhoA signaling[J]. Acta Pharmaceutica Sinica B, 2020, 10(6): 987-1003

TIMP1 preserves the blood-brain barrier through interacting with CD63/integrin β1 complex and regulating downstream FAK/RhoA signaling
Jingshu Tang, Yuying Kang, Longjian Huang, Lei Wu, Ying Penga
State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China
Blood-brain barrier (BBB) breakdown and the associated microvascular hyperpermeability are hallmark features of several neurological disorders, including traumatic brain injury (TBI). However, there is no viable therapeutic strategy to rescue BBB function. Tissue inhibitor of metalloproteinase-1 (TIMP1) has been considered to be beneficial for vascular integrity, but the molecular mechanisms underlying the functions of TIMP1 remain elusive. Here, we report that TIMP1 executes a protective role on neuroprotective function via ameliorating BBB disruption in mice with experimental TBI. In human brain microvessel endothelial cells (HBMECs) exposed to hypoxia and inflammation injury, the recombinant TIMP1 (rTIMP1) treatment maintained integrity of junctional proteins and trans-endothelial tightness. Mechanistically, TIMP1 interacts with CD63/integrin β1 complex and activates downstream FAK signaling, leading to attenuation of RhoA activation and F-actin depolymerization for endothelial cells structure stabilization. Notably, these effects depend on CD63/integrin β1 complex, instead of the MMP-inhibitory function. Together, our results identified a novel MMP-independent function of TIMP1 in regulating endothelial barrier integrity. Therapeutic interventions targeting TIMP1 and its downstream signaling may be beneficial to protect BBB function following brain injury and neurological disorders.
Key words:    issue inhibitor of metalloproteinase-1    Blood-brain barrier    Junctional proteins    CD63    Integrin β1   
Received: 2019-12-25     Revised: 2020-01-22
DOI: 10.1016/j.apsb.2020.02.015
Funds: This project was supported by the grants from National Natural Science Foundation of China (Nos. 81872855 and 81673420), CAMS Innovation Fund for Medical Sciences (No. 2017-I2M-2-004, China), National Science and Technology Major Project on Major New Drug Innovation of China (2018ZX09711001-003-005 and 2018ZX09711001-003-009), Fundamental Research Funds for the Central Universities (3332019070, China) and Disciplines construction project (20190200802, China).
Corresponding author: Ying Peng     Email:ypeng@imm.ac.cn
Author description:
PDF(KB) Free
Jingshu Tang
Yuying Kang
Longjian Huang
Lei Wu
Ying Penga

1. Alluri H, Wiggins-Dohlvik K, Davis ML, Huang JH, Tharakan B. Blood-brain barrier dysfunction following traumatic brain injury. Metab Brain Dis 2015;30:1093-104.
2. Cheng C, Yu Z, Zhao S, Liao Z, Xing C, Jiang Y, et al. Thrombospondin-1 gene deficiency worsens the neurological outcomes of traumatic brain injury in mice. Int J Med Sci 2017;14:927-36.
3. Neuwelt EA, Bauer B, Fahlke C, Fricker G, Iadecola C, Janigro D, et al. Engaging neuroscience to advance translational research in brain barrier biology. Nat Rev Neurosci 2011;12:169-82.
4. Korczyn AD. Vascular parkinsonism-characteristics, pathogenesis and treatment. Nat Rev Neurol 2015;11:319-26.
5. Drouin-Ouellet J, Sawiak SJ, Cisbani G, Lagace M, Kuan WL, SaintPierre M, et al. Cerebrovascular and blood-brain barrier impairments in Huntington's disease:potential implications for its pathophysiology. Ann Neurol 2015;78:160-77.
6. Zenaro E, Piacentino G, Constantin G. The blood-brain barrier in Alzheimer's disease. Neurobiol Dis 2017;107:41-56.
7. Ortiz GG, Pacheco-Moises FP, Macias-Islas MA, Flores-Alvarado LJ, Mireles-Ramirez MA, Gonzalez-Renovato ED, et al. Role of the blood-brain barrier in multiple sclerosis. Arch Med Res 2014;45:687-97.
8. Dietrich W, Erbguth F. Increased intracranial pressure and brain edema. Med Klin Intensivmed Notfallmed 2013;108:157-69.
9. Thal SC, Neuhaus W. The blood-brain barrier as a target in traumatic brain injury treatment. Arch Med Res 2014;45:698-710.
10. Obermeier B, Verma A, Ransohoff RM. The blood-brain barrier. Handb Clin Neurol 2016;133:39-59.
11. Woessner Jr JF. Matrix metalloproteinases and their inhibitors in connective tissue remodeling. FASEB J 1991;5:2145-54.
12. Ries C. Cytokine functions of TIMP-1. Cell Mol Life Sci 2014;71:659-72.
13. Ichiyama T, Siba P, Suarkia D, Takasu T, Miki K, Kira R, et al. Serum levels of matrix metalloproteinase-9 and tissue inhibitors of metalloproteinases 1 in subacute sclerosing panencephalitis. J Neurol Sci 2007;252:45-8.
14. Palus M, Zampachova E, Elsterova J, Ruzek D. Serum matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 levels in patients with tick-borne encephalitis. J Infect 2014;68:165-9.
15. Li DD, Song JN, Huang H, Guo XY, An JY, Zhang M, et al. The roles of MMP-9/TIMP-1 in cerebral edema following experimental acute cerebral infarction in rats. Neurosci Lett 2013;550:168-72.
16. Fujimoto M, Takagi Y, Aoki T, Hayase M, Marumo T, Gomi M, et al. Tissue inhibitor of metalloproteinases protect blood-brain barrier disruption in focal cerebral ischemia. J Cerebr Blood Flow Metabol 2008;28:1674-85.
17. Magnoni S, Baker A, Thomson S, Jordan G, George SJ, McColl BW, et al. Neuroprotective effect of adenoviral-mediated gene transfer of TIMP-1 and -2 in ischemic brain injury. Gene Ther 2007;14:621-5.
18. Mandel ER, Uchida C, Nwadozi E, Makki A, Haas TL. Tissue inhibitor of metalloproteinase 1 influences vascular adaptations to chronic alterations in blood flow. J Cell Physiol 2017;232:831-41.
19. Takawale A, Zhang P, Patel VB, Wang X, Oudit G, Kassiri Z. Tissue inhibitor of matrix metalloproteinase-1 promotes myocardial fibrosis by mediating CD63-integrin beta1 interaction. Hypertension 2017;69:1092-103.
20. Gomis-Ruth FX, Maskos K, Betz M, Bergner A, Huber R, Suzuki K, et al. Mechanism of inhibition of the human matrix metalloproteinase stromelysin-1 by TIMP-1. Nature 1997;389:77-81.
21. Jung KK, Liu XW, Chirco R, Fridman R, Kim HR. Identification of CD63 as a tissue inhibitor of metalloproteinase-1 interacting cell surface protein. EMBO J 2006;25:3934-42.
22. Cui H, Seubert B, Stahl E, Dietz H, Reuning U, Moreno-Leon L, et al. Tissue inhibitor of metalloproteinases-1 induces a pro-tumourigenic increase of miR-210 in lung adenocarcinoma cells and their exosomes. Oncogene 2015;34:3640-50.
23. Forte D, Salvestrini V, Corradi G, Rossi L, Catani L, Lemoli RM, et al. The tissue inhibitor of metalloproteinases-1 (TIMP-1) promotes survival and migration of acute myeloid leukemia cells through CD63/PI3K/Akt/p21 signaling. Oncotarget 2017;8:2261-74.
24. Lee SY, Kim JM, Cho SY, Kim HS, Shin HS, Jeon JY, et al. TIMP-1 modulates chemotaxis of human neural stem cells through CD63 and integrin signalling. Biochem J 2014;459:565-76.
25. Althoff GE, Wolfer DP, Timmesfeld N, Kanzler B, Schrewe H, Pagenstecher A. Long-term expression of tissue-inhibitor of matrix metalloproteinase-1 in the murine central nervous system does not alter the morphological and behavioral phenotype but alleviates the course of experimental allergic encephalomyelitis. Am J Pathol 2010;177:840-53.
26. Crocker SJ, Whitmire JK, Frausto RF, Chertboonmuang P, Soloway PD, Whitton JL, et al. Persistent macrophage/microglial activation and myelin disruption after experimental autoimmune encephalomyelitis in tissue inhibitor of metalloproteinase-1-deficient mice. Am J Pathol 2006;169:2104-16.
27. Stilley JA, Sharpe-Timms KL. TIMP1 contributes to ovarian anomalies in both an MMP-dependent and -independent manner in a rat model. Biol Reprod 2012;86:1-10.
28. Caley MP, Martins VL, O'Toole EA. Metalloproteinases and wound healing. Adv Wound Care (New Rochelle) 2015;4:225-34.
29. Chen SF, Hsu CW, Huang WH, Wang JY. Post-injury baicalein improves histological and functional outcomes and reduces inflammatory cytokines after experimental traumatic brain injury. Br J Pharmacol 2008;155:1279-96.
30. Shaw KE, Bondi CO, Light SH, Massimino LA, McAloon RL, Monaco CM, et al. Donepezil is ineffective in promoting motor and cognitive benefits after controlled cortical impact injury in male rats. J Neurotrauma 2013;30:557-64.
31. Fox GB, Fan L, Levasseur RA, Faden AI. Sustained sensory/motor and cognitive deficits with neuronal apoptosis following controlled cortical impact brain injury in the mouse. J Neurotrauma 1998;15:599-614.
32. Zhou X, Wu Y, Ye L, Wang Y, Zhang K, Wang L, et al. Aspirin alleviates endothelial gap junction dysfunction through inhibition of NLRP3 inflammasome activation in LPS-induced vascular injury. Acta Pharm Sin B 2019;9:711-23.
33. Kim KH, Burkhart K, Chen P, Frevert CW, Randolph-Habecker J, Hackman RC, et al. Tissue inhibitor of metalloproteinase-1 deficiency amplifies acute lung injury in bleomycin-exposed mice. Am J Respir Cell Mol Biol 2005;33:271-9.
34. Yang Y, Rosenberg GA. Blood-brain barrier breakdown in acute and chronic cerebrovascular disease. Stroke 2011;42:3323-8.
35. Yan EB, Satgunaseelan L, Paul E, Bye N, Nguyen P, Agyapomaa D, et al. Post-traumatic hypoxia is associated with prolonged cerebral cytokine production, higher serum biomarker levels, and poor outcome in patients with severe traumatic brain injury. J Neurotrauma 2014;31:618-29.
36. Murray KN, Parry-Jones AR, Allan SM. Interleukin-1 and acute brain injury. Front Cell Neurosci 2015;9:1-17.
37. O'Shea M, Willenbrock F, Williamson RA, Cockett MI, Freedman RB, Reynolds JJ, et al. Site-directed mutations that alter the inhibitory activity of the tissue inhibitor of metalloproteinases-1:importance of the N-terminal region between cysteine 3 and cysteine 13. Biochemistry 1992;31:10146-52.
38. Mehta D, Tiruppathi C, Sandoval R, Minshall RD, Holinstat M, Malik AB. Modulatory role of focal adhesion kinase in regulating human pulmonary arterial endothelial barrier function. J Physiol 2002; 539:779-89.
39. Schmidt TT, Tauseef M, Yue L, Bonini MG, Gothert J, Shen TL, et al. Conditional deletion of FAK in mice endothelium disrupts lung vascular barrier function due to destabilization of RhoA and Rac1 activities. Am J Physiol Lung Cell Mol Physiol 2013;305:L291-300.
40. Kim JG, Islam R, Cho JY, Jeong H, Cap KC, Park Y, et al. Regulation of RhoA GTPase and various transcription factors in the RhoA pathway. J Cell Physiol 2018;233:6381-92.
41. Burridge K, Wittchen ES. The tension mounts:stress fibers as forcegenerating mechanotransducers. J Cell Biol 2013;200:9-19.
42. Shi Y, Zhang L, Pu H, Mao L, Hu X, Jiang X, et al. Rapid endothelial cytoskeletal reorganization enables early blood-brain barrier disruption and long-term ischaemic reperfusion brain injury. Nat Commun 2016;7:10523.
43. Zhao Z, Nelson AR, Betsholtz C, Zlokovic BV. Establishment and dysfunction of the blood-brain barrier. Cell 2015;163:1064-78.
44. Obermeier B, Daneman R, Ransohoff RM. Development, maintenance and disruption of the blood-brain barrier. Nat Med 2013;19:1584-96.
45. Pla-Navarro I, Bevan D, Hajihosseini MK, Lee M, Gavrilovic J. Interplay between metalloproteinases and cell signalling in blood brain barrier integrity. Histol Histopathol 2018;33:1253-70.
46. Wang X, Jung J, Asahi M, Chwang W, Russo L, Moskowitz MA, et al. Effects of matrix metalloproteinase-9 gene knock-out on morphological and motor outcomes after traumatic brain injury. J Neurosci 2000; 20:7037-42.
47. Asahi M, Wang X, Mori T, Sumii T, Jung JC, Moskowitz MA, et al. Effects of matrix metalloproteinase-9 gene knock-out on the proteolysis of blood-brain barrier and white matter components after cerebral ischemia. J Neurosci 2001;21:7724-32.
48. Chen F, Radisky ES, Das P, Batra J, Hata T, Hori T, et al. TIMP-1 attenuates blood-brain barrier permeability in mice with acute liver failure. J Cerebr Blood Flow Metabol 2013;33:1041-9.
49. Chodobski A, Zink BJ, Szmydynger-Chodobska J. Blood-brain barrier pathophysiology in traumatic brain injury. Transl Stroke Res 2011; 2:492-516.
50. Boulday G, Fitau J, Coupel S, Soulillou JP, Charreau B. Exogenous tissue inhibitor of metalloproteinase-1 promotes endothelial cell survival through activation of the phosphatidylinositol 3-kinase/Akt pathway. Ann N Y Acad Sci 2004;1030:28-36.
51. Murphy FR, Issa R, Zhou X, Ratnarajah S, Nagase H, Arthur MJ, et al. Inhibition of apoptosis of activated hepatic stellate cells by tissue inhibitor of metalloproteinase-1 is mediated via effects on matrix metalloproteinase inhibition:implications for reversibility of liver fibrosis. J Biol Chem 2002;277:11069-76.
52. Schlaepfer DD, Mitra SK. Multiple connections link FAK to cell motility and invasion. Curr Opin Genet Dev 2004;14:92-101.
53. Abedi H, Zachary I. Vascular endothelial growth factor stimulates tyrosine phosphorylation and recruitment to new focal adhesions of focal adhesion kinase and paxillin in endothelial cells. J Biol Chem 1997;272:15442-51.
54. Carbajal JM, Gratrix ML, Yu CH, Schaeffer Jr RC. ROCK mediates thrombin's endothelial barrier dysfunction. Am J Physiol Cell Physiol 2000;279:C195-204.
55. Vepa S, Scribner WM, Parinandi NL, English D, Garcia JG, Natarajan V. Hydrogen peroxide stimulates tyrosine phosphorylation of focal adhesion kinase in vascular endothelial cells. Am J Physiol 1999;277:L150-8.
56. Holinstat M, Knezevic N, Broman M, Samarel AM, Malik AB, Mehta D. Suppression of RhoA activity by focal adhesion kinaseinduced activation of p190RhoGAP:role in regulation of endothelial permeability. J Biol Chem 2006;281:2296-305.
57. Zieseniss A. Hypoxia and the modulation of the actin cytoskeletondemerging interrelations. Hypoxia (Auckl) 2014;2:11-21.
58. Schnoor M, Garcia Ponce A, Vadillo E, Pelayo R, Rossaint J, Zarbock A. Actin dynamics in the regulation of endothelial barrier functions and neutrophil recruitment during endotoxemia and sepsis. Cell Mol Life Sci 2017;74:1985-97.
59. Komarova Y, Malik AB. Regulation of endothelial permeability via paracellular and transcellular transport pathways. Annu Rev Physiol 2010;72:463-93.
60. Giannotta M, Trani M, Dejana E. VE-cadherin and endothelial adherens junctions:active guardians of vascular integrity. Dev Cell 2013;26:441-54.
61. Abu Taha A, Schnittler HJ. Dynamics between actin and the VEcadherin/catenin complex:novel aspects of the ARP2/3 complex in regulation of endothelial junctions. Cell Adhes Migrat 2014;8:125-35.
62. Taddei A, Giampietro C, Conti A, Orsenigo F, Breviario F, Pirazzoli V, et al. Endothelial adherens junctions control tight junctions by VEcadherin-mediated upregulation of claudin-5. Nat Cell Biol 2008;10:923-34.
63. Meister S, Storck SE, Hameister E, Behl C, Weggen S, Clement AM, et al. Expression of the ALS-causing variant hSOD1(G93A) leads to an impaired integrity and altered regulation of claudin-5 expression in an in vitro bloodespinal cord barrier model. J Cerebr Blood Flow Metabol 2015;35:1112-21.
64. Park JC, Baik SH, Han SH, Cho HJ, Choi H, Kim HJ, et al. Annexin A1 restores Abeta1-42-induced blood-brain barrier disruption through the inhibition of RhoAeROCK signaling pathway. Aging Cell 2017; 16:149-61.
65. Gopalakrishnan S, Raman N, Atkinson SJ, Marrs JA. Rho GTPase signaling regulates tight junction assembly and protects tight junctions during ATP depletion. Am J Physiol 1998;275:C798-809.
66. Murakami T, Felinski EA, Antonetti DA. Occludin phosphorylation and ubiquitination regulate tight junction trafficking and vascular endothelial growth factor-induced permeability. J Biol Chem 2009; 284:21036-46.
67. Yang Y, Rosenberg GA. Matrix metalloproteinases as therapeutic targets for stroke. Brain Res 2015;1623:30-8.
Similar articles: