Inhibiting MMP13 Attenuates Deep Vein Thrombosis in a Mouse Model by Reducing the Expression of Pdpn

Abstract

Objective

Matrix metalloproteinase 13 (MMP13) is an extracellular matrix protease that affects the progression of atherosclerotic plaques and arterial thrombi by degrading collagens, modifying protein structures and regulating inflammatory responses, but its role in deep vein thrombosis (DVT) has not been determined. The purpose of this study was to investigate the potential effects of MMP13 and MMP13-related genes on the formation of DVT.

Methods

We altered the expression level of MMP13 in vivo and conducted a transcriptome study to examine the expression and relationship between MMP13 and MMP13-related genes in a mouse model of DVT. After screening genes possibly related to MMP13 in DVT mice, the expression levels of candidate genes in human umbilical vein endothelial cells (HUVECs) and the venous wall were evaluated. The effect of MMP13 on platelet aggregation in HUVECs was investigated in vitro.

Results

Among the differentially expressed genes, interleukin 1 beta, podoplanin (Pdpn), and factor VIII von Willebrand factor (F8VWF) were selected for analysis in mice. When MMP13 was inhibited, the expression level of PDPN decreased significantly in vitro. In HUVECs, overexpression of MMP13 led to an increase in the expression level of PDPN and induced platelet aggregation, while transfection of PDPN-siRNA weakened the ability of MMP13 to increase platelet aggregation.

Conclusions

Inhibiting the expression of MMP13 could reduce the burden of DVT in mice. The mechanism involves downregulating the expression of Pdpn through MMP13, which could provide a novel gene target for DVT diagnosis and treatment.

Keywords

deep vein thrombosis / matrix metalloproteinase 13 / podoplanin

References

[1] TritschlerT, KraaijpoelN, Le GalG, et al.. Venous Thromboembolism: Advances in Diagnosis and Treatment. JAMA, 2018, 320(15): 1583-1594

[2] KrugerPC, EikelboomJW, DouketisJD, et al.. Deep vein thrombosis: update on diagnosis and management. Med J Aust, 2019, 210(11): 516-524

[3] MukhopadhyayS, JohnsonTA, DuruN, et al.. Fibrinolysis and Inflammation in Venous Thrombus Resolution. Front Immunol, 2019, 10: 1348

[4] BranchfordBR, CarpenterSL. The Role of Inflammation in Venous Thromboembolism. Front Pediatr, 2018, 6: 142

[5] FischettiF, TedescoF. Cross-talk between the complement system and endothelial cells in physiologic conditions and in vascular diseases. Autoimmunity, 2006, 39(5): 417-428

[6] KudoY, IizukaS, YoshidaM, et al.. Matrix metalloproteinase-13 (MMP-13) directly and indirectly promotes tumor angiogenesis. J Biol Chem, 2012, 287(46): 38716-38728

[7] MauroA, BuscemiM, GerbinoA. Immunohistochemical and transcriptional expression of matrix metalloproteinases in full-term human umbilical cord and human umbilical vein endothelial cells. J Mol Histol, 2010, 41(6): 367-377

[8] ZuckerS, PeiD, JianC, et al.. Membrane type-matrix metalloproteinases (MT-MMP). Current Topics in Develop Biol, 2003, 54: 1-74

[9] ToutouzasK, SynetosA, NikolaouC, et al.. Matrix metalloproteinases and vulnerable atheromatous plaque. Curr Top Med Chem, 2012, 12(10): 1166-1180

[10] HowesJM, KnäuperV, MalcorJD, et al.. Cleavage by MMP-13 renders VWF unable to bind to collagen but increases its platelet reactivity. J Thromb Haemost, 2020, 18(4): 942-954

[11] HowesJM, PughN, HamaiaSW, et al.. MMP-13 binds to platelet receptors αIIbβ3 and GPVI and impairs aggregation and thrombus formation. Res Pract Thromb Haemost, 2018, 2(2): 370-379

[12] LanH, TongZ, JiaoY, et al.. Deep Vein Thrombosis Is Facilitated by Endothelial-Derived Extracellular Vesicles via the PDI-GRP94-GPIIb/IIIa Pathway in Mice. J Clin Med, 2023, 12(13): 4265

[13] ZhangYB, LiW, YaoLQ, et al.. Expression changes and roles of matrix metalloproteinases in a rat model of traumatic deep vein thrombosis. Chin J Traumatol, 2010, 13(3): 188-192

[14] RosenbergGA, YangY. Vasogenic edema due to tight junction disruption by matrix metalloproteinases in cerebral ischemia. Neurosurg Focus, 2007, 22(5): E44

[15] QuillardT, TesmenitskyY, CroceK, et al.. Selective inhibition of matrix metalloproteinase-13 increases collagen content of established mouse atherosclerosis. Arterioscler Thromb Vasc Biol, 2011, 31(11): 2464-2472

[16] McQuibbanGA, GongJH, WongJP, et al.. Matrix metalloproteinase processing of monocyte chemoattractant proteins generates CC chemokine receptor antagonists with anti-inflammatory properties in vivo. Blood, 2002, 100(4): 1160-1167

[17] HuveneersS, DanenEH. Adhesion signaling–crosstalk between integrins, Src and Rho. J Cell Sci, 2009, 122: 1059-1069 Pt 8)

[18] RodriguesSF, GrangerDN. Blood cells and endothelial barrier function. Tissue Barriers, 2015, 3(1–2): e978720

[19] PayneH, PonomaryovT, WatsonSP, et al.. Mice with a deficiency in CLEC-2 are protected against deep vein thrombosis. Blood, 2017, 129(14): 2013-2020

[20] ShindoK, AishimaS, OhuchidaK, et al.. Podoplanin expression in cancer-associated fibroblasts enhances tumor progression of invasive ductal carcinoma of the pancreas. Mol Cancer, 2013, 12(1): 168

[21] ZhouJ, ZhaoX, XieS, et al.. Transcriptome analysis of Klf15-mediated inhibitory functions in a mouse deep venous thrombosis model. Int J Mol Med, 2020, 45(6): 1735-1752

[22] MurinS, MarelichGP, ArroligaAC, et al.. Hereditary thrombophilia and venous thromboembolism. Am J Respir Crit Care Med, 1998, 158(5Pt1): 1369-1373

[23] XiangQ, WangZ, MuG, et al.. Genetic variants influenced the risk of bleeding and pharmacodynamics of rivaroxaban in patients with nonvalvular atrial fibrillation: A multicentre prospective cohort study. Clin Transl Med, 2023, 13(5): e1263

[24] FuJ, LiS, FengR, et al.. Multiple myeloma-derived MMP-13 mediates osteoclast fusogenesis and osteolytic disease. J Clin Invest, 2016, 126(5): 1759-1772

[25] KhalilSK, SaeedL, FadulA, et al.. DVT as the Initial Presentation of Multiple Myeloma: A Rare Case Report and Literature Review. Cureus, 2023, 15(7): e42600

[26] LiS, PritchardDM, YuLG. Regulation and Function of Matrix Metalloproteinase-13 in Cancer Progression and Metastasis. Cancers (Basel), 2022, 14(13): 3263

[27] EkimH, KutayV, TuncerM, et al.. Management of primary venous aneurysms. Saudi Med J, 2004, 25(3): 303-307

[28] WestbyD, GhoneimBM, NolanF, et al.. Varicose veins as a risk factor for venous thromboembolism in arthroplasty patients: Meta-analysis. Phlebology, 2023, 38(3): 150-156

[29] IrwinC, SynnA, KraissL, et al.. Metalloproteinase expression in venous aneurysms. J Vasc Surg, 2008, 48(5): 1278-1285

[30] SerralheiroP, NovaisA, CairrãoE, et al.. Variability of MMP/TIMP and TGF-β1 Receptors throughout the Clinical Progression of Chronic Venous Disease. Int J Mol Sci, 2017, 19(1): 6

[31] QuillardT, AraújoHA, FranckG, et al.. Matrix metalloproteinase-13 predominates over matrix metalloproteinase-8 as the functional interstitial collagenase in mouse atheromata. Arterioscler Thromb Vasc Biol, 2014, 34(6): 1179-1186

[32] DeguchiJO, AikawaE, LibbyP, et al.. Matrix metalloproteinase-13/collagenase-3 deletion promotes collagen accumulation and organization in mouse atherosclerotic plaques. Circulation, 2005, 112(17): 2708-2715

[33] HowesJM, PughN, KnäuperV, et al.. Modified platelet deposition on matrix metalloproteinase 13 digested collagen I. J Thromb Haemost, 2015, 13(12): 2253-2259

[34] HuQ, EckerM. Overview of MMP-13 as a Promising Target for the Treatment of Osteoarthritis. Int J Mol Sci, 2021, 22(4): 1742

[35] SasanoT, Gonzalez-DelgadoR, MuñozNM, et al.. Podoplanin promotes tumor growth, platelet aggregation, and venous thrombosis in murine models of ovarian cancer. J Thromb Haemost, 2022, 20(1): 104-114

[36] Suzuki-InoueK, TsukijiN, ShiraiT, et al.. Platelet CLEC-2: Roles Beyond Hemostasis. Semin Thromb Hemost, 2018, 44(2): 126-134

[37] RiedlJ, PreusserM, NazariPM, et al.. Podoplanin expression in primary brain tumors induces platelet aggregation and increases risk of venous throm-boembolism. Blood, 2017, 129(13): 1831-1839