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23-02-2024 | Phlebothrombosis

Therapeutic implication of MicroRNA-320a antagonist in attenuating blood clots formed during venous thrombosis

Authors: Swati Srivastava, Iti Garg, Nilanjana Ghosh, Rajeev Varshney

Published in: Journal of Thrombosis and Thrombolysis | Issue 4/2024

Abstract

Venous thrombosis (VT) is a complex multi-factorial disease and a major health concern worldwide. Its clinical implications include deep vein thrombosis (DVT) and pulmonary embolism (PE). VT pathogenesis involves intricate interplay of various coagulants and anti-coagulants. Growing evidences from epidemiological studies have shown that many non-coding microRNAs play significant regulatory role in VT pathogenesis by modulating expressions of large number of gene involved in blood coagulation. Present study aimed to investigate the effect of human micro RNA (hsa-miR)-320a antagonist on thrombus formation in VT. Surgery was performed on Sprague–Dawley (SD) rats, wherein the inferior vena cava (IVC) was ligated to introduce DVT. Animals were divided into four groups (n = 5 in each group); Sham controls (Sham), IVC ligated-DVT (DVT), IVC ligated-DVT + transfection reagent (DVT-NC) and IVC ligated-DVT + miR320a antagonist (DVT-miR-320a antagonist). IVC was dissected after 6 h and 24 h of surgery to estimate thrombus weight and coagulatory parameters such as levels of D-dimer, clotting time and bleeding time. Also, ELISA based biochemical assays were formed to assess toxicity of miRNA antagonist in animals. Our experimental analysis demonstrated that there was a marked reduction in size of thrombus in hsa-miR-320a antagonist treated animals, both at 6 h and 24 h. There was a marked reduction in D-dimer levels in hsa-miR-320a antagonist treated animals. Also, blood clotting time was delayed and bleeding time was increased significantly in hsa-miR-320a antagonist treated rats compared to the non-treated and Sham rats. There was no sign of toxicity in treated group compared to control animals. Hsa-miR-320a antagonist could be promising therapeutic target for management of VT.

ISTH Steering Committee for World Thrombosis Day (2014) Thrombosis: a major contributor to the global disease burden. J Thromb Haemost 12(10):1580–1590. https://doi.org/10.1111/jth.12698CrossRef

Naess IA, Christiansen SC, Romundstad P, Cannegieter SC, Rosendaal FR, Hammerstrøm J (2007) Incidence and mortality of venous thrombosis: a population-based study. J Thromb Haemost 5(4):692–699. https://doi.org/10.1111/j.1538-7836.2007.02450.xCrossRefPubMed

Schulman S, Lindmarker P, Holmström M, Lärfars G, Carlsson A, Nicol P, Svensson E, Ljungberg B, Viering S, Nordlander S, Leijd B, Jahed K, Hjorth M, Linder O, Beckman M (2006) Post-thrombotic syndrome, recurrence, and death 10 years after the first episode of venous thromboembolism treated with warfarin for 6 weeks or 6 months. J Thromb Haemost 4(4):734–742. https://doi.org/10.1111/j.1538-7836.2006.01795.xCrossRefPubMed

Diaz JA, Obi A, Myers DD, Wrobleski SK, Henke PK, Mackman N, Wakefield TW (2012) Critical review of mouse models of venous thrombosis. Arterioscler Thromb Vasc Biol 32:556–562PubMedPubMedCentralCrossRef

Heit JA, O’Fallon WM, Petterson TM, Lohse CM, Silverstein MD, Mohr DN, Melton LJ (2002) Relative impact of risk factors for deep vein thrombosis and pulmonary embolism: a population-based study. Arch Intern Med 162:1245–1248PubMedCrossRef

Zwicker I et al (2009) Tumor-derived tissue factor-bearing microparticles are associated with venous thromboembolic events in malignancy. Clin Cancer Res 15:6830–6840PubMedPubMedCentralCrossRef

Zhou X et al (2010) Incidence and risk factors of venous thromboembolic events in lymphoma. Am J Med 123:935–941PubMedCrossRef

Demers C et al (1998) Incidence of venographically proved deep vein thrombosis after knee arthroscopy. Arch Intern Med 158:47–50PubMedCrossRef

Chan MY, Andreotti F, Becker RC (2008) Hypercoagulable states in cardiovascular disease. Circulation 118:2286–2297PubMedCrossRef

10.

Anand AC, Jha SK, Saha A, Sharma V, Adya CM (2001) Thrombosis as a complication of extended stay at high altitude. Natl Med J India 14(4):197–201PubMed

11.

Khalil KF, Saeed W (2010) Pulmonary embolism in soldiers serving at high altitude. J Coll Physicians Surg Pak 20:7

12.

Kumar S (2006) High altitude induced deep venous thrombosis: a study of 28 cases. Indian J Surg 68:84–88

13.

Rao KS (2006) Other medical illnesses aggravated by high altitude. In: Anand AC, Narula AS, Kakkar R, Kalra R (eds) Textbook of environmental emergencies. Department of Internal Medicine Armed Forces Medical College, Pune, pp 26–29

14.

Singh I, Chohan IS (1972) Blood coagulation changes at high altitude predisposing to pulmonary hypertension. Br Heart J 34:611–617PubMedPubMedCentralCrossRef

15.

Brill A, Fuchs TA, Chauhan AK et al (2011) von Willebrand factor-mediated platelet adhesion is critical for deep vein thrombosis in mouse models. Blood 117:1400–1407. https://doi.org/10.1182/blood-2010-05-287623CrossRefPubMedPubMedCentral

16.

Mammen EF (1992) Pathogenesis of venous thrombosis. Chest. https://doi.org/10.1378/chest.102.6_supplement.640sCrossRefPubMed

17.

Sevitt S (1974) The structure and growth of valve-pocket thrombi in femoral veins. J Clin Pathol 27:517–528. https://doi.org/10.1136/jcp.27.7.517CrossRefPubMedPubMedCentral

18.

Brühl MLV, Stark K, Steinhart A et al (2012) Monocytes, neutrophils, and platelets cooperate to initiate and propagate venous thrombosis in mice in vivo. J Exp Med 209:819–835. https://doi.org/10.1084/jem.20112322CrossRef

19.

Schönfelder T, Jäckel S, Wenzel P (2017) Mouse models of deep vein thrombosis. Gefasschirurgie 22:28–33. https://doi.org/10.1007/s00772-016-0227-6CrossRefPubMed

20.

Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–297PubMedCrossRef

21.

Iwakawa HO, Tomari Y (2015) The functions of microRNAs: mRNA decay and translational repression. Trends Cell Biol 25(11):651–665PubMedCrossRef

22.

Zhu W, Qian J, Ma L, Ma P, Yang F, Shu Y (2017) MiR-346 suppresses cell proliferation through SMYD3 dependent approach in hepatocellular carcinoma. Oncotarget 8(39):65218–65229. https://doi.org/10.18632/oncotarget.18060CrossRefPubMedPubMedCentral

23.

Bartel DP (2009) MicroRNAs: target recognition and regulatory functions. Cell 136:215–233PubMedPubMedCentralCrossRef

24.

Armand-Labit V, Pradines A (2017) Circulating cell-free MicroRNAs as clinical cancer biomarkers. Biomol Concepts 8:61–81PubMedCrossRef

25.

Loosen SH, Schueller F, Trautwein C et al (2017) Role of circulating microRNAs in liver diseases. World J Hepatol 9:586–594PubMedPubMedCentralCrossRef

26.

Rayner K, Dimmeler S, Calin GA et al (2014) Novel biomarkers for acute myocardial infarction: is microRNA the new kid on the block? ClinChem 60:812–817

27.

Liu X, Cheng Y, Zhang S, Lin Y, Yang J, Zhang C (2009) A necessary role of miR-221 and miR-222 in vascular smooth muscle cell proliferation and neointimal hyperplasia. Circ Res 104:476–487PubMedPubMedCentralCrossRef

28.

Tan M, Yan HB, Li JN et al (2016) Thrombin stimulated platelet-derived exosomes inhibit platelet-derived growth factor receptor-beta expression vascular smooth muscle cells. Cell Physiol Biochem 38:2348–2365PubMedCrossRef

29.

Weber C, Schober A, Zernecke A (2010) Micrornas in arterial remodelling, inflammation and atherosclerosis. Curr Drug Targets 11:950–956PubMedCrossRef

30.

Hembrom AA, Srivastava S, Garg I, Kumar B (2020) MicroRNAs in venous thrombo-embolism. Clin Chim Acta 504:66–72. https://doi.org/10.1016/j.cca.2020.01.034CrossRefPubMed

31.

Hembrom AA, Ghosh N, Kumar V, Garg I, Ganju L, Srivastava S (2022) Panel of regulatory miRNAs for blood coagulation under normoxic and hypoxic conditions. Def Life Sci J 7(1):17–26CrossRef

32.

Srivastava S, Garg I, Kumari B, Rai C, Singh Y, Kumar V, Yanamandra U, Singh J, Bansal A, Kumar B (2019) Diagnostic potential of circulating micro RNA hsa-miR-320 in patients of high altitude induced deep vein thrombosis: an Indian study. Gene Rep. 17:100550CrossRef

33.

Srivastava S, Garg I, Kumar B (2019) An In-Silico argument for Micro-RNAs showing pivotal role in susceptibility towards high altitude induced venous thrombo-embolism (HA-VTE). Int J Gen Sci 6(1):1–11

34.

Sahu A, Jha PK, Prabhakar A, Singh HD, Gupta N, Chatterjee T, Tyagi T, Sharma S, Kumari B, Singh S, Nair V, Goel S, Ashraf MZ (2017) MicroRNA-145 impedes thrombus formation via targeting tissue factor in venous thrombosis. EBioMedicine 26:175–186PubMedPubMedCentralCrossRef

35.

Sahu A, Jha PK, Prabhakar A, Singh HD, Gupta N, Chatterjee T, Tyagi T, Sharma S, Kumari B, Singh S et al (2017) MicroRNA-145 impedes thrombus formation via targeting tissue factor in venous thrombosis. EBioMedicine 26:175–186PubMedPubMedCentralCrossRef

36.

Tyagi T, Ahmad S, Gupta N, Sahu A, Ahmad Y, Nair V, Chatterjee T, Bajaj N, Sengupta S, Ganju L, Singh SB, Ashraf MZ (2013) Altered expression of platelet proteins and calpain activity mediate hypoxia-induced prothrombotic phenotype. Blood. https://doi.org/10.1182/blood-2013-05-501924CrossRefPubMed

37.

Gupta N, Sahu A, Prabhakar A, Chatterjee T, Tyagi T, Kumari B, Khan N, Nair V, Bajaj N, Sharma M, Ashraf MZ (2017) Activation of NLRP3 inflammasome complex potentiates venous thrombosis in response to hypoxia. Proc Natl Acad Sci U S A 114(18):4763–4768. https://doi.org/10.1073/pnas.1620458114CrossRefPubMedPubMedCentral

38.

Wang W, Zhu X, Du X, Xu A, Yuan X, Zhan Y, Liu M, Wang S (2019) MiR-150 promotes angiogensis and proliferation of endothelial progenitor cells in deep venous thrombosis by targeting SRCIN1. Microvasc Res 123:35–41PubMedCrossRef

39.

Wang X, Sundquist K, Svensson PJ et al (2019) Association of recurrent venous thromboembolism and circulating microRNAs. Clin Epigenet 11:28CrossRef

40.

Rodriguez-Rius A, Lopez S, Martinez-Perez A, Souto JC, Soria JM (2020) Identification of a plasma MicroRNA Profile associated with venous thrombosis. Arterioscler, Thromb, Vasc Biol 40:1392–1399PubMedCrossRef

41.

Rodriguez-Rius A, Lopez S, Martinez-Perez A, Souto JC, Soria JM (2020) Identification of a plasma MicroRNA profile associated with venous thrombosis. Arterioscler Thromb Vasc Biol 40(5):1392–1399. https://doi.org/10.1161/ATVBAHA.120.314092CrossRefPubMed

42.

Mussbacher M, Krammer TL, Heber S et al (2020) Impact of anticoagulation and sample processing on the quantification of human blood-derived microRNA signatures. Cells 9(8):E1915. https://doi.org/10.3390/cells9081915CrossRef

43.

Jiang Z, Ma J, Wang Q, Wu F, Ping J, Ming L (2018) Combination of circulating miRNA-320a/b and D-dimer improves diagnostic accuracy in deep vein thrombosis patients. Med Sci Monit 6(24):2031–2037CrossRef

44.

Starikova I, Jamaly S, Sorrentino A, Blondal T, Latysheva N, Sovershaev M, Hansen JB (2015) Differential expression of plasma miRNAs in patients with unprovoked venous thromboembolism and healthy control individuals. Thromb Res 136(3):566–572PubMedCrossRef

45.

Thomas PD, Ebert D, Muruganujan A, Mushayahama T, Albou L-P, Mi H (2022) PANTHER: making genome-scale phylogenetics accessible to all. Protein Soc 31(1):8–22. https://doi.org/10.1002/pro.421CrossRef

46.

Kloosterman WP, Plasterk RH (2006) The diverse functions of microRNAs in animal development and disease. Dev Cell 11:441–450. https://doi.org/10.1016/j.devcel.2006.09.009CrossRefPubMed

47.

Paul P, Chakraborty A, Sarkar D, Langthasa M, Rahman M, Bari M, Singha RS, Malakar AK, Chakraborty S (2018) Interplay between miRNAs and human diseases. J Cell Physiol 233:2007–2018. https://doi.org/10.1002/jcp.25854CrossRefPubMed

48.

Rupaimoole R, Slack FJ (2017) MicroRNA therapeutics: towards a new era for the management of cancer and other diseases. Nat Rev Drug Discov 16:203–222. https://doi.org/10.1038/nrd.2016.246CrossRefPubMed

49.

Van Rooij E, Kauppinen S (2014) Development of microRNA therapeutics is coming of age. EMBO Mol Med 6:851–864. https://doi.org/10.15252/emmm.201100899CrossRefPubMedPubMedCentral

50.

Morelli VM, Brækkan SK, Hansen JB (2020) Role of microRNAs in venous thromboembolism. Int J Mol Sci 21(7):2602PubMedPubMedCentralCrossRef

51.

Denby L, Ramdas V, Lu R, Conway BR, Grant JS, Dickinson B, Aurora AB, McClure JD, Kipgen D, Delles C, van Rooij E, Baker AH (2014) MicroRNA-214 antagonism protects against renal fibrosis. J Am Soc Nephrol 25(1):65–80PubMedCrossRef

52.

Zhang H, Qu Y, Wang A (2018) Antagonist targeting microRNA-146a protects against lithium-pilocarpine-induced status epilepticus in rats by nuclear factor-κB pathway. Mol Med Rep 17:5356–5361PubMed

53.

Hum C, Loiselle J, Ahmed N, Shaw TA, Toudic C, Pezacki JP (2021) MicroRNA mimics or inhibitors as antiviral therapeutic approaches against COVID-19. Drugs 81(5):517–531PubMedPubMedCentralCrossRef

54.

Sun L, Li X, Li Q, Wang L, Li J, Shu C (2021) Multiple arterial and venous thromboembolism in a male patient with hereditary protein C deficiency. Medicine (Baltimore) 100(15):e25575. https://doi.org/10.1097/MD.0000000000025575CrossRefPubMed

55.

Du X, Hong L, Sun L, Sang H, Qian A, Li W, Zhuang H, Liang H, Song D, Li C et al (2019) miR-21 induces endothelial progenitor cells proliferation and angiogenesis via targeting FASLG and is a potential prognostic marker in deep venous thrombosis. J Transl Med 17:270PubMedPubMedCentralCrossRef

56.

Kong L, Du X, Hu N, Li W, Wang W, Wei S, Zhuang H, Li X, Li C (2016) Downregulation of let-7e-5p contributes to endothelial progenitor cell dysfunction in deep vein thrombosis via targeting FASLG. Thromb Res 138:30–36PubMedCrossRef

57.

Kong L, Hu N, Du X, Wang W, Chen H, Li W, Wei S, Zhuang H, Li X, Li C (2016) Upregulation of miR-483-3p contributes to endothelial progenitor cells dysfunction in deep vein thrombosis patients via SRF. J Transl Med 14:23PubMedPubMedCentralCrossRef

58.

Meng Q, Wang W, Yu X, Li W, Kong L, Qian A, Li C, Li X (2015) Upregulation of MicroRNA-126 contributes to endothelial progenitor cell function in deep vein thrombosis via its target PIK3R2. J Cell Biochem 116:1613–1623. https://doi.org/10.1002/jcb.25115CrossRefPubMed

59.

Wang W, Li C, Li W, Kong L, Qian A, Hu N, Meng Q, Li X (2014) MiR-150 enhances the motility of EPCs in vitro and promotes EPCs homing and thrombus resolving in vivo. Thromb Res 133:590–598PubMedCrossRef

60.

Sun S, Chai S, Zhang F, Lu L (2020) Overexpressed microRNA-103a-3p inhibits acute lower-extremity deep venous thrombosis via inhibition of CXCL12. IUBMB Life 72(3):492–504. https://doi.org/10.1002/iub.2168CrossRefPubMed

61.

Sun S, Chai S, Zhang F, Lu L (2020) Overexpressed microRNA-103a-3p inhibits acute lower-extremity deep venous thrombosis via inhibition of CXCL12. IUBMB Life 72:492–504. https://doi.org/10.1002/iub.2168CrossRefPubMed

62.

Zhang Y, Zhang Z, Wei R, Miao X, Sun S, Liang G, Chu C, Zhao L, Zhu X, Guo Q et al (2020) IL (Interleukin)-6 contributes to deep vein thrombosis and is negatively regulated by miR-338-5p. Arterioscler Thromb Vasc Biol 40:323–334PubMedCrossRef

63.

McCreight J, Schneider S, Wilburn D, Swanson W (2017) Evolution of microRNA in primates. PLoS ONE 12:e0176596. https://doi.org/10.1371/journal.pone.0176596CrossRefPubMedPubMedCentral

64.

Chen X, Gao S, Zhao Z, Liang G, Kong J, Feng X (2020) MicroRNA-320d regulates tumor growth and invasion by promoting FoxM1 and predicts poor outcome in gastric cardiac adenocarcinoma. Cell Biosci 10:80. https://doi.org/10.1186/s13578-020-00439-7CrossRefPubMedPubMedCentral

65.

Fang Z, Tang J, Bai Y, Lin H, You H, Jin H, Lin L, You P, Li J, Dai Z et al (2015) Plasma levels of microRNA-24, microRNA-320a, and microRNA-423-5p are potential biomarkers for colorectal carcinoma. J Exp Clin Cancer Res 34:86. https://doi.org/10.1186/s13046-015-0198-6CrossRefPubMedPubMedCentral

66.

Hong H, Zhu H, Zhao S, Wang K, Zhang N, Tian Y, Li Y, Wang Y, Lv X, Wei T et al (2019) The novel circCLK3/miR-320a/FoxM1 axis promotes cervical cancer progression. Cell Death Dis 10:950. https://doi.org/10.1038/s41419-019-2183-zCrossRefPubMedPubMedCentral

67.

Lv Q, Du H, Liu Y, Huang Y, Wang G, Zhang X, Chen S, Zhou H (2017) Low expression of microRNA-320b correlates with tumorigenesis and unfavorable prognosis in glioma. Oncol Rep 38:959–966. https://doi.org/10.3892/or.2017.5762CrossRefPubMed

68.

Thibord F, Munsch G, Perret C, Suchon P, Roux M, Ibrahim-Kosta M, Goumidi L, Deleuze JF, Morange PE, Trégouët DA (2020) Bayesian network analysis of plasma microRNA sequencing data in patients with venous thrombosis. Eur Heart J Suppl 22(Suppl C):C34–C45PubMedPubMedCentralCrossRef

69.

Diener C, Keller A, Meese E (2022) Emerging concepts of miRNA therapeutics: from cells to clinic. Trends Genet 38(6):613–626PubMedCrossRef

70.

Nagalla S, Shaw C, Kong X et al (2011) Platelet microRNA-mRNA coexpression profiles correlate with platelet reactivity. Blood 117(19):5189–5197. https://doi.org/10.1182/blood-2010-09-299719CrossRefPubMedPubMedCentral

Title: Therapeutic implication of MicroRNA-320a antagonist in attenuating blood clots formed during venous thrombosis
Authors: Swati Srivastava
Iti Garg
Nilanjana Ghosh
Rajeev Varshney
Publication date: 23-02-2024
Publisher: Springer US
Keywords: Phlebothrombosis
Venous Thrombosis
Venous Thrombosis
Published in: Journal of Thrombosis and Thrombolysis / Issue 4/2024
Print ISSN: 0929-5305
Electronic ISSN: 1573-742X
DOI: https://doi.org/10.1007/s11239-024-02947-6

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